https://gitlab.com/tgwgraham/papa_paper_plasmids
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Tip revision: 984803582c0023a9fe975ec6d41be3d723c2737b authored by Thomas Graham on 08 July 2022, 01:06:31 UTC
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Tip revision: 9848035
ptg839-ef1alpha-3xflag-halo-snapf-60mer.gb
LOCUS       pTG839_EF1alpha-3xFlag- 9165 bp ds-DNA     circular     20-NOV-2021
DEFINITION  .
KEYWORDS    "creator:Gina Dailey - Darzacq/Tjian Lab" "marker:AmpR, PuroR"     
FEATURES             Location/Qualifiers
     primer          complement(108..126)
                     /label="oTG1552_pBS_diffusion_substrate_300bp"
                     /note="sequence: ctctagcttcccggcaaca"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     CDS             complement(650..718)
                     /label="AmpR"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     promoter        complement(719..823)
                     /label="AmpR promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer          complement(802..823)
                     /label="oTG1179_FRT_AmpR_5’"
                     /note="sequence: CTATACTTTCTAGAGAATAGGAACTTCcgcggaacccctatttgtttat"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     primer          complement(821..839)
                     /label="oTG1553_pBS_diffusion_substrate_1kb"
                     /note="sequence: ttttcggggaaatgtgcgc"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     rep_origin      complement(849..1303)
                     /label="f1 ori"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          1441..1460
                     /label="oTG1336_M13F_extended"
                     /note="sequence: GTTGTAAAACGACGGCCAGT"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     primer_bind     1443..1460
                     /label="M13-fwd"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer_bind     1444..1460
                     /label="M13 fwd"
                     /ApEinfo_revcolor=#a020f0
                     /ApEinfo_fwdcolor=#a020f0
     misc_feature    1590..1602
                     /label="PB Terminal repeat"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    1637..1652
                     /label="IR (inverted repeat) truncated?"
                     /ApEinfo_revcolor=#339966
                     /ApEinfo_fwdcolor=#339966
     promoter        2071..2570
                     /label="PGK promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     CDS             2592..3191
                     /label="PuroR"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     polyA_signal    3224..3448
                     /label="bGH poly(A) signal"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     promoter        3488..3699
                     /label="EF-1α core promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     promoter        3488..3699
                     /label="EF-1α core promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     LTR             3712..3980
                     /label="5' LTR (truncated)"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     LTR             3712..3980
                     /label="5' LTR (truncated)"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     primer          3991..4010
                     /label="oTG1576_3xFlag-Halo_upstream_F"
                     /note="sequence: CCGGCGCCTACTCTAGCCAC"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     CDS             4012..4014
                     /label="ATG"
                     /ApEinfo_revcolor=#993366
                     /ApEinfo_fwdcolor=#993366
     CDS             4012..6218
                     /label="Translation 4012-6218"
     CDS             4015..4080
                     /label="3xFLAG"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     primer          complement(4057..4081)
                     /label="oTG1615_Flag_RT2_R"
                     /note="sequence: CTAGCttgctagtgcttgtcatcgtcatccttgtaatc"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          complement(4057..4081)
                     /label="oTG1613_Flag_RT1_R"
                     /note="sequence: CTAGCttgctattgcttgtcatcgtcatccttgtaatc"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          complement(4057..4081)
                     /label="oTG1617_Flag_RT3_R"
                     /note="sequence: CTAGCgccctattgcttgtcatcgtcatccttgtaatc"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          4089..4114
                     /label="oTG945_60xHalo_F"
                     /note="sequence: CCGGCTCCGGCAGCGGTAGCATGGCAGAAATCGGTACTGGCTTTC"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     primer          4090..4110
                     /label="oTG1582_SNAPf_linker_Halo_F"
                     /note="sequence: TGGGCAAGCCTGGGCTGGGTggtagtggtaccggttctATGGCAGAAATCGGTACTGGC"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer          4090..4110
                     /label="oTG778_5'Halo"
                     /note="sequence: ATGGCAGAAATCGGTACTGGC"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          4090..4110
                     /label="oTG1573_GSGSKCK_Halo_F"
                     /note="sequence: GGTAGTGGTAGTAAGTGTAAAATGGCAGAAATCGGTACTGGC"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer          4090..4110
                     /label="oTG898_H10_Halo_F"
                     /note="sequence: TTCTCACCATCACCATCACCACCACCACCACCATATGGCAGAAATCGGTACTGGC"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     CDS             4090..4980
                     /label="HaloTag®"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     primer          4093..4113
                     /label="oTG1144_inVitro_60xHalo_F"
                     /note="sequence: GGCTCCGGCAGCGGTAGCGCAGAAATCGGTACTGGCTTT"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     primer          4096..4118
                     /label="oTG1280_60mer_linker_Halo_F"
                     /note="sequence: GGAGTGGTAGCGGCAGTATTAAGgaaatcggtactggctttccatt"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          4096..4118
                     /label="oTG995_Halo_linker_F"
                     /note="sequence: GATCTGGAGGTTCAGGAGGCAGCGGTGGAAGTGAAATCGGTACTGGCTTTCCATT"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer          complement(4101..4120)
                     /label="oTG1172_Halo_int_R2"
                     /note="sequence: CGAATGGAAAGCCAGTACCG"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer          complement(4106..4125)
                     /label="oTG1214_Halo_int_R_75"
                     /note="sequence: GGGGTCGAATGGAAAGCCAG"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          complement(4129..4148)
                     /label="oTG1096_Halo_int_R"
                     /note="sequence: TCGCCCAGGACTTCCACATA"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          complement(4150..4169)
                     /label="oTG1173_Halo_int_R3"
                     /note="sequence: ACATCGACGTAGTGCATGCG"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     primer          4311..4332
                     /label="oTG1513_Halo_qPCR_F1"
                     /note="sequence: ACCAGACCTGGGTTATTTCTTC"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     primer          4451..4470
                     /label="oTG1208_Halo_int_F"
                     /note="sequence: AGCGCGTCAAAGGTATTGCA"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer          complement(4471..4490)
                     /label="oTG1514_Halo_qPCR_R1"
                     /note="sequence: GGGCGGATGAACTCCATAAA"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer          complement(4565..4587)
                     /label="oTG1674_Halo_V167C_R"
                     /note="sequence: GTTCTGATCGATGATCAGCTTGC"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          complement(4565..4587)
                     /label="oTG1691_Halo_V167C_R"
                     /note="sequence: GTTCTGATCGATGATCAGCTTGC"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          complement(4574..4599)
                     /label="oTG1676_Halo_G171C_R"
                     /note="sequence: CTCGATAAAAACGTTCTGATCGATGA"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          complement(4574..4599)
                     /label="oTG1693_Halo_G171C_R"
                     /note="sequence: CTCGATAAAAACGTTCTGATCGATGA"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          4590..4610
                     /label="oTG1690_Halo_V167C_F"
                     /note="sequence: tgtTTTATCGAGGGTACGCTGCC"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          4590..4610
                     /label="oTG1673_Halo_V167C_F"
                     /note="sequence: tgtTTTATCGAGGGTACGCTGCC"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          complement(4592..4611)
                     /label="oTG1678_Halo_M175C_R"
                     /note="sequence: CGGCAGCGTACCCTCGATAA"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          complement(4592..4611)
                     /label="oTG1695_Halo_M175C_R"
                     /note="sequence: CGGCAGCGTACCCTCGATAA"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          4601..4620
                     /label="oTG1692_Halo_G171C_F"
                     /note="sequence: tgtACGCTGCCGATGGGTGTC"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          4601..4620
                     /label="oTG1675_Halo_G171C_F"
                     /note="sequence: tgtACGCTGCCGATGGGTGTC"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          4615..4632
                     /label="oTG1677_Halo_M175C_F"
                     /note="sequence: tgtGGTGTCGTCCGCCCGCTG"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          4615..4632
                     /label="oTG1694_Halo_M175C_F"
                     /note="sequence: tgtGGTGTCGTCCGCCCGCTG"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          complement(4629..4650)
                     /label="oTG1207_Halo_int_R"
                     /note="sequence: GTCCATCTCGACTTCAGTCAGC"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer          4636..4657
                     /label="oTG1515_Halo_qPCR_F2"
                     /note="sequence: GAAGTCGAGATGGACCATTACC"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     primer          complement(4758..4777)
                     /label="oTG1516_Halo_qPCR_R2"
                     /note="sequence: GCAGCCAGTCCATGTATTCT"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          4871..4890
                     /label="oTG1095_Halo_int_F"
                     /note="sequence: ACTGCAAGGCTGTGGACATC"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer          4898..4920
                     /label="oTG979_3'_Halo_seq"
                     /note="sequence: GTCTGAATCTGCTGCAAGAAGAC"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer          complement(4948..4965)
                     /label="oTG1312_Halo_tubUTR_R"
                     /note="sequence: cgcgacgcttagtgtgccgcTTACGTCGACAGCCAGCGCGC"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer          complement(4960..4979)
                     /label="oTG1261_ParB2_Halo_stop_R"
                     /note="sequence: AGGTTCCTTCACAAAGATCCTCTAGAttaACCGGAAATCTCGAGCGTCGA"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer          complement(4960..4979)
                     /label="oTG1261_ParB2_Halo_stop_R"
                     /note="sequence: AGGTTCCTTCACAAAGATCCTCTAGAttaACCGGAAATCTCGAGCGTCGA"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer          complement(4961..4980)
                     /label="oTG793_3'Halo_R"
                     /note="sequence: GCCGGAAATCTCGAGCGTCG"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer          complement(4961..4980)
                     /label="oTG779_Halo-stop_pETSUMO"
                     /note="sequence: GCTCCAATTGGGTACCCATGGAGCCGGAAATCTCGAGCGTCG"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     primer          complement(4961..4982)
                     /label="oTG1583_Halo_noNLS_R"
                     /note="sequence: TGATTATGATCAGTTATCTAGATCCGGTGGATCCgccggaaatctcgagcgtcg"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    4972..4991
                     /label="Gibson overhang"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          4972..4991
                     /label="oTG1618_SNAP-3xNLS_gBlock_F"
                     /note="sequence: ATTTCCGGCggtagtggtac"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     misc_feature    4999..5544
                     /label="SNAPf"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     CDS             4999..5544
                     /label="Translation 4999-5544"
     primer          5550..5569
                     /label="oTG1683_3xNLS-P2A-T2A_KpnI_F"
                     /note="sequence: atttccggcggtagtggtaccTCAGGAAGTGGAGATGGCG"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          5577..5626
                     /label="oTG1620_HaloSNAP60mer-T2AP2A-SNAP_F"
                     /note="sequence: GATCAATGGACGGGCAGATgctagcATGAAAATGGAAGAGCTGTTCAAGA"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          5601..5626
                     /label="oTG909_60mer_pcDNA_F"
                     /note="sequence: CCACTAGTCCAGTGTGGTGGCCACCATGAAAATGGAAGAGCTGTTCAAGA"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer          5601..5626
                     /label="oTG1622_HaloSNAP3xN-TP2A-60merSNAP_F"
                     /note="sequence: gaaaatcccggcccaGgtaccATGAAAATGGAAGAGCTGTTCAAGA"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     misc_feature    5602..6216
                     /label="60mer"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer          5605..5626
                     /label="oTG936_60xGFP_F"
                     /note="sequence: AAAATGGAAGAGCTGTTCAAGA"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer          5605..5630
                     /label="oTG977_Linker_ctGFP_F"
                     /note="sequence: GTTCAGGAGGCAGCGGTGGAAGTAAAATGGAAGAGCTGTTCAAGAAACA"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     primer          complement(6198..6216)
                     /label="oTG1621_HaloSNAP60mer-T2AP2A-SNAP_R"
                     /note="sequence: ttgtggcgccgctgcccatatgTTCGGTGCAGCCACGAATC"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          complement(6198..6216)
                     /label="oTG1623_HaloSNAP60mer-TP2A-SNAP_R"
                     /note="sequence: tccacttccgccaccggtaccTTCGGTGCAGCCACGAATC"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     primer          complement(6198..6218)
                     /label="oTG1623_HaloSNAP60mer-TP2A-SNAP_R"
                     /note="sequence: tccacttccgccaccggtaccTTCGGTGCAGCCACGAATC"
                     /ApEinfo_revcolor=#f7977a
                     /ApEinfo_fwdcolor=#f7977a
     CDS             6217..6231
                     /label="Translation 6217-6231"
     CDS             6236..6247
                     /label="Translation 6236-6247"
     misc_feature    6237..6247
                     /label="stop codons"
                     /ApEinfo_revcolor=#ff0000
                     /ApEinfo_fwdcolor=#ff0000
     misc_feature    6324..6324
                     /label="add_before"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     CDS             6324..6341
                     /label="MCS"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     polyA_signal    6457..6578
                     /label="SV40 poly(A) signal"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer          complement(6550..6578)
                     /label="oTG885_SV40_pA_F"
                     /note="sequence: TAAGATACATTGATGAGTTTGGACAAACC"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    complement(7338..7356)
                     /label="IR (inverted repeat)"
                     /ApEinfo_revcolor=#339966
                     /ApEinfo_fwdcolor=#339966
     misc_feature    complement(7360..7372)
                     /label="PB Terminal repeat"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     primer_bind     complement(7863..7879)
                     /label="M13 rev"
                     /ApEinfo_revcolor=#a020f0
                     /ApEinfo_fwdcolor=#a020f0
     protein_bind    7887..7903
                     /label="lac operator"
                     /ApEinfo_revcolor=#31849b
                     /ApEinfo_fwdcolor=#31849b
     promoter        complement(7911..7917)
                     /label="lac promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     promoter        complement(7918..7935)
                     /label="lac promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer          complement(7929..7950)
                     /label="oTG980_Vector-Fwd-GA"
                     /note="sequence: ccggatgacagtgacgaggagaactgaACCCCAGGCTTTacacTTTATG"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     promoter        complement(7936..7941)
                     /label="lac promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     protein_bind    7956..7977
                     /label="CAP binding site"
                     /ApEinfo_revcolor=#31849b
                     /ApEinfo_fwdcolor=#31849b
     primer          8008..8028
                     /label="oTG1156_mCherry-LacI-Rev"
                     /note="sequence: AAGCTTCCATTCGGGAAACCTGTCGtgCCAG"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer          8028..8048
                     /label="oTG1089_Rev-LacI-NLS-NOSTOP-GA"
                     /note="sequence: AACCTTCCTCTTCTTCTTAGGAGGCTGCATTAATGAATCGGCCA"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     primer          8028..8049
                     /label="oTG1084_Rev-LacI-GA"
                     /note="sequence: TCAAACCTTCCTCTtcTTcTTaGgagGCTGCATTAATGAATCGGCCAA"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     primer          8085..8102
                     /label="oTG775_pET_BbvCI_insert_seq"
                     /note="sequence: TTCCGCTTCCTCGCTCAC"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     rep_origin      complement(8265..8852)
                     /label="ori"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          8906..8929
                     /label="oTG1180_AmpR_seq"
                     /note="sequence: ACGTTAAGGGATTTTGGTCATGAG"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer          complement(8992..9022)
                     /label="oTG1177_FRT_AmpR_3’flanking"
                     /note="sequence: CTATACTTTCTAGAGAATAGGAACTTCctgtcagaccaagtttactcatatatacttt"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     primer          complement(8992..9022)
                     /label="oTG1554_pBS_diffusion_substrate_3kb"
                     /note="sequence: ctgtcagaccaagtttactcatatatacttt"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     primer          8993..9022
                     /label="oTG1182_E75_AmpR_F"
                     /note="sequence: gactaactgccactcgcagcgcgcagatcgAAGTATATATGAGTAAACTTGGTCTGACAG"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     primer          9022..9046
                     /label="oTG1549_pBS_849_adapter"
                     /note="sequence: GCTGTGTCTTATTACACACAGTCGttaccaatgcttaatcagtgaggc"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     CDS             complement(join(9023..9165,1..649))
                     /label="AmpR"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     primer          9023..9046
                     /label="oTG1176_FRT_AmpR_3’"
                     /note="sequence: CTATTCTCTAGAAAGTATAGGAACTTCttaccaatgcttaatcagtgaggc"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          complement(9069..9091)
                     /label="oTG1550_pBS_diffusion_substrate_100bp"
                     /note="sequence: caggcaactatggatgaacgaaa"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
ORIGIN
        1 ACCCACGCTC ACCGGTTCCA GATTTATCAG CAATAAACCA GCCAGCCGGA AGGGCCGAGC
       61 GCAGAAGTGG TCCTGCAACT TTATCCGCCT CCATCCAGTC TATTAATTGT TGCCGGGAAG
      121 CTAGAGTAAG TAGTTCGCCA GTTAATAGTT TGCGCAACGT TGTTGCCATT GCTACAGGCA
      181 TCGTGGTGTC ACGCTCGTCG TTTGGTATGG CTTCATTCAG CTCCGGTTCC CAACGATCAA
      241 GGCGAGTTAC ATGATCCCCC ATGTTGTGCA AAAAAGCGGT TAGCTCCTTC GGTCCTCCGA
      301 TCGTTGTCAG AAGTAAGTTG GCCGCAGTGT TATCACTCAT GGTTATGGCA GCACTGCATA
      361 ATTCTCTTAC TGTCATGCCA TCCGTAAGAT GCTTTTCTGT GACTGGTGAG TACTCAACCA
      421 AGTCATTCTG AGAATAGTGT ATGCGGCGAC CGAGTTGCTC TTGCCCGGCG TCAATACGGG
      481 ATAATACCGC GCCACATAGC AGAACTTTAA AAGTGCTCAT CATTGGAAAA CGTTCTTCGG
      541 GGCGAAAACT CTCAAGGATC TTACCGCTGT TGAGATCCAG TTCGATGTAA CCCACTCGTG
      601 CACCCAACTG ATCTTCAGCA TCTTTTACTT TCACCAGCGT TTCTGGGTGA GCAAAAACAG
      661 GAAGGCAAAA TGCCGCAAAA AAGGGAATAA GGGCGACACG GAAATGTTGA ATACTCATAC
      721 TCTTCCTTTT TCAATATTAT TGAAGCATTT ATCAGGGTTA TTGTCTCATG AGCGGATACA
      781 TATTTGAATG TATTTAGAAA AATAAACAAA TAGGGGTTCC GCGCACATTT CCCCGAAAAG
      841 TGCCACCTAA ATTGTAAGCG TTAATATTTT GTTAAAATTC GCGTTAAATT TTTGTTAAAT
      901 CAGCTCATTT TTTAACCAAT AGGCCGAAAT CGGCAAAATC CCTTATAAAT CAAAAGAATA
      961 GACCGAGATA GGGTTGAGTG TTGTTCCAGT TTGGAACAAG AGTCCACTAT TAAAGAACGT
     1021 GGACTCCAAC GTCAAAGGCG AAAAACCGTC TATCAGGGCG ATGGCCCACT ACGTGAACCA
     1081 TCACCCTAAT CAAGTTTTTT GGGGTCGAGG TGCCGTAAAG CACTAAATCG GAACCCTAAA
     1141 GGGAGCCCCC GATTTAGAGC TTGACGGGGA AAGCCGGCGA ACGTGGCGAG AAAGGAAGGG
     1201 AAGAAAGCGA AAGGAGCGGG CGCTAGGGCG CTGGCAAGTG TAGCGGTCAC GCTGCGCGTA
     1261 ACCACCACAC CCGCCGCGCT TAATGCGCCG CTACAGGGCG CGTCCCATTC GCCATTCAGG
     1321 CTGCGCAACT GTTGGGAAGG GCGATCGGTG CGGGCCTCTT CGCTATTACG CCAGCTGGCG
     1381 AAAGGGGGAT GTGCTGCAAG GCGATTAAGT TGGGTAACGC CAGGGTTTCC CAGTCACGAC
     1441 GTTGTAAAAC GACGGCCAGT GAGCGCGCCT CGTTCATTCA CGTTTTTGAA CCCGTGGAGG
     1501 ACGGGCAGAC TCGCGGTGCA AATGTGTTTT ACAGCGTGAT GGAGCAGATG AAGATGCTCG
     1561 ACACGCTGCA GAACACGCAG CTAGATTAAC CCTAGAAAGA TAATCATATT GTGACGTACG
     1621 TTAAAGATAA TCATGTGTAA AATTGACGCA TGTGTTTTAT CGGTCTGTAT ATCGAGGTTT
     1681 ATTTATTAAT TTGAATAGAT ATTAAGTTTT ATTATATTTA CACTTACATA CTAATAATAA
     1741 ATTCAACAAA CAATTTATTT ATGTTTATTT ATTTATTAAA AAAAACAAAA ACTCAAAATT
     1801 TCTTCTATAA AGTAACAAAA CTTTTATGAG GGACAGCCCC CCCCCAAAGC CCCCAGGGAT
     1861 GTAATTACGT CCCTCCCCCG CTAGGGGGCA GCAGCGAGCC GCCCGGGGCT CCGCTCCGGT
     1921 CCGGCGCTCC CCCCGCATCC CCGAGCCGGC AGCGTGCGGG GACAGCCCGG GCACGGGGAA
     1981 GGTGGCACGG GATCGCTTTC CTCTGAACGC TTCTCGCTGC TCTTTGAGCC TGCAGACACC
     2041 TGGGGGGATA CGGGGAAAAG GCGAACGCGT gggtagggga ggcgcttttc ccaaggcagt
     2101 ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc
     2161 ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct
     2221 tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt
     2281 cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca
     2341 ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct
     2401 tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc
     2461 tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa
     2521 agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcagcc
     2581 caAGCTTCAC CATGACCGAG TACAAGCCCA CGGTGCGCCT CGCCACCCGC GACGACGTCC
     2641 CCAGGGCCGT ACGCACCCTC GCCGCCGCGT TCGCCGACTA CCCCGCCACG CGCCACACCG
     2701 TCGATCCGGA CCGCCACATC GAGCGGGTCA CCGAGCTGCA AGAACTCTTC CTCACGCGCG
     2761 TCGGGCTCGA CATCGGCAAG GTGTGGGTCG CGGACGACGG CGCCGCGGTG GCGGTCTGGA
     2821 CCACGCCGGA GAGCGTCGAA GCGGGGGCGG TGTTCGCCGA GATCGGCCCG CGCATGGCCG
     2881 AGTTGAGCGG TTCCCGGCTG GCCGCGCAGC AACAGATGGA AGGCCTCCTG GCGCCGCACC
     2941 GGCCCAAGGA GCCCGCGTGG TTCCTGGCCA CCGTCGGCGT CTCGCCCGAC CACCAGGGCA
     3001 AGGGTCTGGG CAGCGCCGTC GTGCTCCCCG GAGTGGAGGC GGCCGAGCGC GCCGGGGTGC
     3061 CCGCCTTCCT GGAGACCTCC GCGCCCCGCA ACCTCCCCTT CTACGAGCGG CTCGGCTTCA
     3121 CCGTCACCGC CGACGTCGAG GTGCCCGAAG GACCGCGCAC CTGGTGCATG ACCCGCAAGC
     3181 CCGGTGCCTG AGCGGCCCGT TTAAACCCGC TGATCAGCCT CGACTGTGCC TTCTAGTTGC
     3241 CAGCCATCTG TTGTTTGCCC CTCCCCCGTG CCTTCCTTGA CCCTGGAAGG TGCCACTCCC
     3301 ACTGTCCTTT CCTAATAAAA TGAGGAAATT GCATCGCATT GTCTGAGTAG GTGTCATTCT
     3361 ATTCTGGGGG GTGGGGTGGG GCAGGACAGC AAGGGGGAGG ATTGGGAAGA CAATAGCAGG
     3421 CATGCTGGGG ATGCGGTGGG CTCTATGGCT TCTGAGGCGG AACGCGATCG CTCCGGTGCC
     3481 CGTCAGTGGG CAGAGCGCAC ATCGCCCACA GTCCCCGAGA AGTTGGGGGG AGGGGTCGGC
     3541 AATTGAACGG GTGCCTAGAG AAGGTGGCGC GGGGTAAACT GGGAAAGTGA TGTCGTGTAC
     3601 TGGCTCCGCC TTTTTCCCGA GGGTGGGGGA GAACCGTATA TAAGTGCAGT AGTCGCCGTG
     3661 AACGTTCTTT TTCGCAACGG GTTTGCCGCC AGAACACAGC TGAAGCTTCG AGGGGCTCGC
     3721 ATCTCTCCTT CACGCGCCCG CCGCCCTACC TGAGGCCGCC ATCCACGCCG GTTGAGTCGC
     3781 GTTCTGCCGC CTCCCGCCTG TGGTGCCTCC TGAACTGCGT CCGCCGTCTA GGTAAGTTTA
     3841 AAGCTCAGGT CGAGACCGGG CCTTTGTCCG GCGCTCCCTT GGAGCCTACC TAGACTCAGC
     3901 CGGCTCTCCA CGCTTTGCCT GACCCTGCTT GCTCAACTCT ACGTCTTTGT TTCGTTTTCT
     3961 GTTCTGCGCC GTTACAGATC CAAGCTGTGA CCGGCGCCTA CTCTAGCCAC CATGGACTAC
     4021 AAAGACCATG ACGGTGATTA TAAAGATCAT GACATCGATT ACAAGGATGA CGATGACAAG
     4081 CTTGGCTCCA TGGCAGAAAT CGGTACTGGC TTTCCATTCG ACCCCCATTA TGTGGAAGTC
     4141 CTGGGCGAGC GCATGCACTA CGTCGATGTT GGTCCGCGCG ATGGCACCCC TGTGCTGTTC
     4201 CTGCACGGTA ACCCGACCTC CTCCTACGTG TGGCGCAACA TCATCCCGCA TGTTGCACCG
     4261 ACCCATCGCT GCATTGCTCC AGACCTGATC GGTATGGGCA AATCCGACAA ACCAGACCTG
     4321 GGTTATTTCT TCGACGACCA CGTCCGCTTC ATGGATGCCT TCATCGAAGC CCTGGGTCTG
     4381 GAAGAGGTCG TCCTGGTCAT TCACGACTGG GGCTCCGCTC TGGGTTTCCA CTGGGCCAAG
     4441 CGCAATCCAG AGCGCGTCAA AGGTATTGCA TTTATGGAGT TCATCCGCCC TATCCCGACC
     4501 TGGGACGAAT GGCCAGAATT TGCCCGCGAG ACCTTCCAGG CCTTCCGCAC CACCGACGTC
     4561 GGCCGCAAGC TGATCATCGA TCAGAACGTT TTTATCGAGG GTACGCTGCC GATGGGTGTC
     4621 GTCCGCCCGC TGACTGAAGT CGAGATGGAC CATTACCGCG AGCCGTTCCT GAATCCTGTT
     4681 GACCGCGAGC CACTGTGGCG CTTCCCAAAC GAGCTGCCAA TCGCCGGTGA GCCAGCGAAC
     4741 ATCGTCGCGC TGGTCGAAGA ATACATGGAC TGGCTGCACC AGTCCCCTGT CCCGAAGCTG
     4801 CTGTTCTGGG GCACCCCAGG CGTTCTGATC CCACCGGCCG AAGCCGCTCG CCTGGCCAAA
     4861 AGCCTGCCTA ACTGCAAGGC TGTGGACATC GGCCCGGGTC TGAATCTGCT GCAAGAAGAC
     4921 AACCCGGACC TGATCGGCAG CGAGATCGCG CGCTGGCTGT CGACGCTCGA GATTTCCGGC
     4981 ggtagtggta caggttctAT GGATAAAGAC TGTGAGATGA AGCGGACAAC CCTTGATTCC
     5041 CCTCTGGGGA AACTGGAGCT CAGTGGGTGC GAACAGGGGC TGCACAGAAT TATATTTCTT
     5101 GGGAAGGGCA CGTCTGCTGC AGATGCTGTC GAGGTCCCAG CCCCGGCTGC CGTGCTTGGT
     5161 GGACCAGAGC CTCTCATGCA AGCTACTGCT TGGCTGAACG CGTATTTCCA CCAACCTGAA
     5221 GCGATAGAAG AGTTCCCTGT TCCTGCTCTG CACCACCCGG TTTTCCAACA GGAATCTTTC
     5281 ACGAGACAAG TATTGTGGAA ACTTCTCAAA GTGGTCAAAT TCGGTGAAGT AATATCCTAT
     5341 TCACACCTGG CAGCTCTCGC CGGGAACCCG GCGGCAACCG CTGCGGTTAA AACTGCACTC
     5401 TCTGGTAATC CTGTACCAAT ACTTATACCG TGCCACAGGG TTGTCCAAGG CGACTTGGAC
     5461 GTTGGAGGGT ATGAAGGGGG CCTCGCTGTG AAAGAGTGGC TTCTCGCGCA CGAAGGTCAT
     5521 CGGCTCGGTA AGCCAGGGCT CGGGGAAATC TCAGGAAGTG GAGATGGCGC CGGATTGATC
     5581 AATGGACGGG CAGATgctag cATGAAAATG GAAGAGCTGT TCAAGAAACA CAAGATCGTT
     5641 GCCGTGCTGC GTGCCAATAG TGTGGAAGAA GCGAAAAAGA AAGCGCTGGC GGTTTTCCTG
     5701 GGCGGCGTTC ATCTGATTGA AATTACCTTT ACCGTGCCGG ATGCGGATAC CGTGATTAAG
     5761 GAACTGAGCT TTCTGAAGGA AATGGGCGCG ATTATTGGTG CGGGCACCGT GACCAGCGTG
     5821 GAGCAGTGCC GTAAAGCGGT GGAAAGTGGC GCCGAATTCA TTGTGAGTCC GCACCTGGAC
     5881 GAGGAAATTA GCCAATTTTG CAAGGAGAAG GGTGTGTTCT ATATGCCAGG CGTTATGACC
     5941 CCGACCGAAC TGGTGAAAGC CATGAAACTG GGCCATACCA TCTTAAAACT GTTTCCGGGT
     6001 GAGGTGGTGG GTCCGCAGTT TGTTAAAGCG ATGAAAGGTC CGTTTCCGAA TGTGAAATTT
     6061 GTGCCAACCG GCGGTGTTAA TCTGGACAAT GTGTGCGAAT GGTTCAAAGC GGGCGTGCTG
     6121 GCCGTGGGCG TGGGCAGCGC GTTAGTGAAA GGCACCCCGG TGGAAGTGGC GGAAAAGGCC
     6181 AAGGCGTTCG TTGAGAAGAT TCGTGGCTGC ACCGAAcaTA GCGGTACCTA GTGATCTAGA
     6241 TAACTGATCA TAATCAGCCA TACCACATTT GTAGAGGTTT TACTTGCTTT AAAAAACCTC
     6301 CCACACCTCC CCCTGAACCT GAACGCGGCC GCtggGTCGA CAATCAACCT CTGGATTACA
     6361 AAATTTGTGA AAGATTGACT GGTATTCTTA ACTATGTTGC TCCTTTTACG CTATGTGGAT
     6421 ACGCTGCTTT AATGCCTTTG TATCATGCGT TAACTAAACT TGTTTATTGC AGCTTATAAT
     6481 GGTTACAAAT AAAGCAATAG CATCACAAAT TTCACAAATA AAGCATTTTT TTCACTGCAT
     6541 TCTAGTTGTG GTTTGTCCAA ACTCATCAAT GTATCTTATC ATGTCTGGAA TTGACTCAAA
     6601 TGATGTCAAT TAGTCTATCA GAAGCTCATC TGGTCTCCCT TCCGGGGGAC AAGACATCCC
     6661 TGTTTAATAT TTAAACAGCA GTGTTCCCAA ACTGGGTTCT TATATCCCTT GCTCTGGTCA
     6721 ACCAGGTTGC AGGGTTTCCT GTCCTCACAG GAACGAAGTC CCTAAAGAAA CAGTGGCAGC
     6781 CAGGTTTAGC CCCGGAATTG ACTGGATTCC TTTTTTAGGG CCCATTGGTA TGGCTTTTTC
     6841 CCCGTATCCC CCCAGGTGTC TGCAGGCTCA AAGAGCAGCG AGAAGCGTTC AGAGGAAAGC
     6901 GATCCCGTGC CACCTTCCCC GTGCCCGGGC TGTCCCCGCA CGCTGCCGGC TCGGGGATGC
     6961 GGGGGGAGCG CCGGACCGGA GCGGAGCCCC GGGCGGCTCG CTGCTGCCCC CTAGCGGGGG
     7021 AGGGACGTAA TTACATCCCT GGGGGCTTTG GGGGGGGGCT GTCCCTGATA TCTATAACAA
     7081 GAAATATATA TATAATAAGT TATCACGTAA GTAGAACATG AAATAACAAT ATAATTATCG
     7141 TATGAGTTAA TCTTAAAGTC ACGTAAAAGA TAATCATGCG TCATTTTGAC TCACGCGGTC
     7201 GTTATAGTTC AAAATCAGTG ACACTTACCG CATTGACAAG CACGCCTCAC GGGAGCTCCA
     7261 AGCGGCGACT GAGATGTCCT AAATGCACAG CGACGGATTC GCGCTATTTA GAAAGAGAGA
     7321 GCAATATTTC AAGAATGCAT GCGTCAATTT TACGCAGACT ATCTTTCTAG GGTTAATCTA
     7381 GCTGCATCAG GATCATATCG TCGGGTCTTT TTTCCGGCTC AGTCATCGCC CAAGCTGGCG
     7441 CTATCTGGGC ATCGGGGAGG AAGAAGCCCG TGCCTTTTCC CGCGAGGTTG AAGCGGCATG
     7501 GAAAGAGTTT GCCGAGGATG ACTGCTGCTG CATTGACGTT GAGCGAAAAC GCACGTTTAC
     7561 CATGATGATT CGGGAAGGTG TGGCCATGCA CGCCTTTAAC GGTGAACTGT TCGTTCAGGC
     7621 CACCTGGGAT ACCAGTTCGT CGCGGCTTTT CCGGACACAG TTCCGGATGG TCAGCCCGAA
     7681 GCGCATCAGC AACCCGAACA ATACCGGCGA CAGCCGGAAC TGCCGTGCCG GTGTGCAGAT
     7741 TAATGACAGC GGTGCGGCGC TGGGATATTA CGTCAGCGAG GACGGGTATC CTGGCTGGAT
     7801 GCCGCAGAAA TGGACATGGA TACCCCGTGA GTTACCCGGC GGGCGCGCTT GGCGTAATCA
     7861 TGGTCATAGC TGTTTCCTGT GTGAAATTGT TATCCGCTCA CAATTCCACA CAACATACGA
     7921 GCCGGAAGCA TAAAGTGTAA AGCCTGGGGT GCCTAATGAG TGAGCTAACT CACATTAATT
     7981 GCGTTGCGCT CACTGCCCGC TTTCCAGTCG GGAAACCTGT CGTGCCAGCT GCATTAATGA
     8041 ATCGGCCAAC GCGCGGGGAG AGGCGGTTTG CGTATTGGGC GCTCTTCCGC TTCCTCGCTC
     8101 ACTGACTCGC TGCGCTCGGT CGTTCGGCTG CGGCGAGCGG TATCAGCTCA CTCAAAGGCG
     8161 GTAATACGGT TATCCACAGA ATCAGGGGAT AACGCAGGAA AGAACATGTG AGCAAAAGGC
     8221 CAGCAAAAGG CCAGGAACCG TAAAAAGGCC GCGTTGCTGG CGTTTTTCCA TAGGCTCCGC
     8281 CCCCCTGACG AGCATCACAA AAATCGACGC TCAAGTCAGA GGTGGCGAAA CCCGACAGGA
     8341 CTATAAAGAT ACCAGGCGTT TCCCCCTGGA AGCTCCCTCG TGCGCTCTCC TGTTCCGACC
     8401 CTGCCGCTTA CCGGATACCT GTCCGCCTTT CTCCCTTCGG GAAGCGTGGC GCTTTCTCAT
     8461 AGCTCACGCT GTAGGTATCT CAGTTCGGTG TAGGTCGTTC GCTCCAAGCT GGGCTGTGTG
     8521 CACGAACCCC CCGTTCAGCC CGACCGCTGC GCCTTATCCG GTAACTATCG TCTTGAGTCC
     8581 AACCCGGTAA GACACGACTT ATCGCCACTG GCAGCAGCCA CTGGTAACAG GATTAGCAGA
     8641 GCGAGGTATG TAGGCGGTGC TACAGAGTTC TTGAAGTGGT GGCCTAACTA CGGCTACACT
     8701 AGAAGAACAG TATTTGGTAT CTGCGCTCTG CTGAAGCCAG TTACCTTCGG AAAAGAGTTG
     8761 GTAGCTCTTG ATCCGGCAAA CAAACCACCG CTGGTAGCGG TGGTTTTTTT GTTTGCAAGC
     8821 AGCAGATTAC GCGCAGAAAA AAAGGATCTC AAGAAGATCC TTTGATCTTT TCTACGGGGT
     8881 CTGACGCTCA GTGGAACGAA AACTCACGTT AAGGGATTTT GGTCATGAGA TTATCAAAAA
     8941 GGATCTTCAC CTAGATCCTT TTAAATTAAA AATGAAGTTT TAAATCAATC TAAAGTATAT
     9001 ATGAGTAAAC TTGGTCTGAC AGTTACCAAT GCTTAATCAG TGAGGCACCT ATCTCAGCGA
     9061 TCTGTCTATT TCGTTCATCC ATAGTTGCCT GACTCCCCGT CGTGTAGATA ACTACGATAC
     9121 GGGAGGGCTT ACCATCTGGC CCCAGTGCTG CAATGATACC GCGAG
//
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