pwFRT-pheS-gat vector (V002137)

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LOCUS       V002137                 4531 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002137
VERSION     V002137
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 4531)
  AUTHORS   Kang Y, Norris MH, Hoang TT.
  TITLE     Knock-out and pull-out recombineering protocols for natural
            transformable Burkholderia thailandensis and Burkholderia
            pseudomallei
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4531)
  AUTHORS   Kang Y, Norris MH, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-OCT-2009) Molecular Biosciences and Biological
            Engineering, University of Hawaii at Manoa, 2538 McCarthy Mall,
            Snyder Hall 207, Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 4531)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4531)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (09-OCT-2009) Molecular Biosciences and Biological Engineering,
            University of Hawaii at Manoa, 2538 McCarthy Mall, Snyder Hall 207,
            Honolulu, HI 96822, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /label="AmpR promoter"
     CDS             201..1058
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1232..1820
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    2108..2129
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2144..2174
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2206..2222
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    2289..2356
                     /note="wild type FRT; Flp recognition target"
     protein_bind    complement(2306..2353)
                     /label="FRT"
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     regulatory      2393..2434
                     /note="PCS12; rpsL promoter from Burkholderia cenocepacia"
                     /regulatory_class="promoter"
     RBS             2445..2467
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2478..3488
                     /gene="pheS"
                     /label="Phenylalanine--tRNA ligase alpha subunit"
                     /note="Phenylalanine--tRNA ligase alpha subunit from
                     Burkholderia pseudomallei (strain 1710b). Accession#:
                     Q3JT08"
     CDS             3527..3967
                     /codon_start=1
                     /gene="gat"
                     /product="glyphosate acetyl transferase"
                     /label="gat"
                     /note="confers resistance to glyphosate"
                     /protein_id="ADO63837.1"
                     /translation="MIEVKPINAEDTYDLRHRVLRPNQPIEACMFESDLTRSAFHLGGF
                     YGGKLISVASFHQAEHSELQGKKQYQLRGVATLEGYREQKAGSSLVKHAEEILRKRGAD
                     MIWCNARTSASGYYRKLGFSEQGEVFDTPPVGPHILMYKRIT"
     gene            3527..3967
                     /gene="gat"
                     /label="gat"
     protein_bind    complement(4023..4070)
                     /label="FRT"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(4137..4153)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"