pFlp-AB7 vector (V006226)

LOCUS       V006226                 7680 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V006226
VERSION     V006226
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7680)
  AUTHORS   Norris MH, Kang Y, Barrett AR, Lu D, Hoang TT.
  TITLE     Glyphosate resistance as a novel non-antibiotic selectable marker in
            the select agent Burkholderia pseudomallei
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7680)
  AUTHORS   Norris MH, Kang Y, Barrett AR, Lu D, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-OCT-2008) Molecular Biology and Bio-Engineering,
            University of Hawaii at Manoa, 2538 McCarthy Mall, Snyder 308,
            Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 7680)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7680)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (14-OCT-2008) Molecular Biology and Bio-Engineering, University of
            Hawaii at Manoa, 2538 McCarthy Mall, Snyder 308, Honolulu, HI 96822,
            USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(232..583)
                     /direction=LEFT
                     /label="pRO1600 oriV"
                     /note="broad-host-range origin of replication from
                     Pseudomonas aeruginosa plasmid pRO1600; requires the
                     pRO1600 Rep protein for replication (West et al., 1994)"
     CDS             597..1427
                     /label="pRO1600 Rep"
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
     primer_bind     1591..1607
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     regulatory      1662..1703
                     /note="PCS12; rpsL promoter from Burkholderia pseudomallei"
                     /regulatory_class="promoter"
     CDS             1739..2749
                     /gene="pheS"
                     /label="Phenylalanine--tRNA ligase alpha subunit"
                     /note="Phenylalanine--tRNA ligase alpha subunit from
                     Burkholderia pseudomallei (strain 1710b). Accession#:
                     Q3JT08"
     CDS             complement(2964..4232)
                     /label="FLP"
                     /note="site-specific recombinase"
     RBS             4236..4244
                     /label="Shine-Dalgarno sequence"
                     /note="full consensus sequence for ribosome-binding sites
                     upstream of start codons in E. coli; complementary to a
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             4333..5043
                     /label="lambda repressor (ts)"
                     /note="temperature-sensitive variant of the phage lambda
                     repressor"
     primer_bind     complement(5335..5351)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5359..5375)
                     /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)."
     promoter        complement(5383..5413)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5428..5449)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     oriT            5833..5941
                     /label="oriT"
                     /note="incP origin of transfer"
     rep_origin      complement(6013..6601)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6921..7361)
                     /codon_start=1
                     /gene="gat"
                     /product="Gat"
                     /label="gat"
                     /note="glyphosate acetyl transferase; confers resistance to
                     glyphosate"
                     /protein_id="ACJ70061.1"
                     /translation="MIEVKPINAEDTYDLRHRVLRPNQPIEACMFESDLTRSAFHLGGF
                     YGGKLISVASFHQAEHSELQGKKQYQLRGVATLEGYREQKAGSSLVKHAEEILRKRGAD
                     MIWCNARTSASGYYRKLGFSEQGEVFDTPPVGPHILMYKRIT"
     gene            complement(6921..7361)
                     /gene="gat"
                     /label="gat"
     RBS             complement(7397..7419)
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      complement(7430..7471)
                     /note="PCS12; rpsL promoter from Burkholderia cenocepacia"
                     /regulatory_class="promoter"