pBT20-Dbla-T7pol vector (V009018)

LOCUS       V009018                 9952 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009018
VERSION     V009018
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9952)
  AUTHORS   Kang Y, Son MS, Hoang TT.
  TITLE     One step engineering of T7-expression strains for protein
            production: increasing the host-range of the T7-expression system
  JOURNAL   Protein Expr. Purif. 55 (2), 325-333 (2007)
   PUBMED   17716915
REFERENCE   2  (bases 1 to 9952)
  AUTHORS   Kang Y, Son MS, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-NOV-2006) Microbiology, University of Hawaii at Manoa,
            2538 The Mall - Snyder 310, Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 9952)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9952)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein
            Expr. Purif."; date: "2007"; volume: "55"; issue: "2"; pages:
            "325-333"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (30-NOV-2006) Microbiology, University of Hawaii at Manoa, 2538 The
            Mall - Snyder 310, Honolulu, HI 96822, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             34..402
                     /label="traJ"
                     /note="oriT-recognizing protein"
     rep_origin      1021..1409
                     /label="R6K gamma ori"
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     repeat_region   1451..1478
                     /rpt_family="Mariner"
     CDS             complement(1549..4197)
                     /note="T7 RNA polymerase from Escherichia phage T7.
                     Accession#: P00573"
                     /label="T7 RNA polymerase"
     CDS             complement(4506..4679)
                     /label="lacZ-alpha"
                     /note="LacZ-alpha fragment of beta-galactosidase"
     misc_feature    complement(4697..4721)
                     /label="lac operator"
                     /note="lac operator"
     protein_bind    4699..4715
                     /label="lac operator"
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(4723..4753)
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    complement(4768..4789)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4805..5884)
                     /label="lacI"
                     /note="lac repressor"
     misc_feature    complement(5965..6012)
                     /note="FRT; Flp-recombination-target"
     protein_bind    complement(5965..6012)
                     /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)."
     promoter        6053..6081
                     /label="Pc promoter"
                     /note="class 1 integron promoter"
     CDS             6270..6800
                     /label="GmR"
                     /note="gentamycin acetyltransferase"
     protein_bind    complement(6933..6980)
                     /label="FRT"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             7243..7695
                     /label="T7 lysozyme"
                     /note="lysozyme from bacteriophage T7"
     repeat_region   7730..7757
                     /rpt_family="Mariner"
     primer_bind     7890..7906
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        7916..7934
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(7997..9043)
                     /codon_start=1
                     /gene="Tnp"
                     /product="Tnp"
                     /label="Tnp"
                     /note="mariner transposase"
                     /protein_id="ABN54776.1"
                     /translation="MEKKEFRVLIKYCFLKGKNTVEAKTWLDNEFPDSAPGKSTIIDWY
                     AKFKRGEMSTEDGERSGRPKEVVTDENIKKIHKMILNDRKMKLIEIAEALKISKERVGH
                     IIHQYLDMRKLCAKWVPRELTFDQKQRRVDDSKRCLQLLTRNTPEFFRRYVTMDETWLH
                     HYTPESNRQSAEWTATGEPSPKRGKTQKSAGKVMASVFWDAHGIIFIDYLEKGKTINSD
                     YYMALLERLKVEIAAKRPHMKKKKVLFHQDNAPCHKSLRTMAKIHELGFELLPHPPYSP
                     DLAPSDFFLFSDLKRMLAGKKFGCNEEVIAETEAYFEAKPKEYYQNGIKKLEGRYNRCI
                     ALEGNYVE"
     gene            complement(7997..9043)
                     /gene="Tnp"
                     /label="Tnp"
     oriT            9844..9952
                     /label="oriT"
                     /note="incP origin of transfer"