p94FLP_asRepL vector (V009694)

LOCUS       V009694                 8046 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V009694
VERSION     V009694
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8046)
  AUTHORS   Al-Hinai MA, Fast AG, Papoutsakis ET.
  TITLE     Novel system for efficient isolation of clostridium double-crossover
            allelic exchange mutants enabling markerless chromosomal gene
            deletions and DNA integration
  JOURNAL   Appl. Environ. Microbiol. 78 (22), 8112-8121 (2012)
   PUBMED   22983967
REFERENCE   2  (bases 1 to 8046)
  AUTHORS   Al-Hinai MA, Papoutsakis ET, Fast AG.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-SEP-2012) Biological Sciences, University of Delaware,
            15 Innovation Way, Newark, DE 19711, USA
REFERENCE   3  (bases 1 to 8046)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8046)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Environ. Microbiol."; date: "2012"; volume: "78"; issue: "22";
            pages: "8112-8121"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (06-SEP-2012) Biological Sciences, University of Delaware, 15
            Innovation Way, Newark, DE 19711, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8046
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     381..397
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(465..905)
                     /codon_start=1
                     /gene="repL"
                     /product="gram positive replication protein"
                     /label="repL"
                     /note="RepL"
                     /protein_id="AFS68390.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     gene            complement(465..905)
                     /gene="repL"
                     /label="repL"
     CDS             complement(1356..2087)
                     /gene="ermC'"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from Bacillus
                     subtilis. Accession#: P13956"
     CDS             complement(2667..3506)
                     /codon_start=1
                     /gene="bgaR"
                     /product="beta-galactosidase regulator"
                     /label="bgaR"
                     /note="BgaR"
                     /protein_id="AFS68392.1"
                     /translation="MQILWKKYVKENFEMNVDECGIEQGIPGLGYNYEVLKNAVIHYVT
                     KGYGTFKFNGKVYNLKQGDIFILLKGMQVEYVASIDDPWEYYWIGFSGSNANEYLNRTS
                     ITNSCVANCEENSKIPQIILNMCEISKTYNPSRSDDILLLKELYSLLYALIEEFPKPFE
                     YKDKELHTYIQDALNFINSNYMHSITVQEIADYVNLSRSYLYKMFIKNLGISPQRYLIN
                     LRMYKATLLLKSTKLPIGEVASSVGYSDSLLFSKTFSKHFSMSPLNYRNNQVNKPSI"
     gene            complement(2667..3506)
                     /gene="bgaR"
                     /label="bgaR"
     gene            3923..4022
                     /gene="repLAS"
                     /label="repLAS"
                     /note="antisense RNA to the repL"
     CDS             4360..5628
                     /label="FLP"
                     /note="site-specific recombinase"
     primer_bind     complement(5825..5841)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5849..5865)
                     /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(5873..5903)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5918..5939)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6227..6815)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6989..7846)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7847..7951)
                     /label="AmpR promoter"