pKOD_mazF vector (V005201)

LOCUS       V005201                 7202 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005201
VERSION     V005201
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7202)
  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 7202)
  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 7202)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7202)
  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..7202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             37..693
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     CDS             1038..1340
                     /label="ccdB"
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(1384..1508)
                     /label="attR2"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1533..1973)
                     /codon_start=1
                     /gene="repL"
                     /product="gram positive replication protein"
                     /label="repL"
                     /note="RepL"
                     /protein_id="AFS68389.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     gene            complement(1533..1973)
                     /gene="repL"
                     /label="repL"
     CDS             complement(2584..3423)
                     /codon_start=1
                     /gene="bgaR"
                     /product="beta-galactosidase regulator"
                     /label="bgaR"
                     /note="BgaR"
                     /protein_id="AFS68386.1"
                     /translation="MQILWKKYVKENFEMNVDECGIEQGIPGLGYNYEVLKNAVIHYVT
                     KGYGTFKFNGKVYNLKQGDIFILLKGMQVEYVASIDDPWEYYWIGFSGSNANEYLNRTS
                     ITNSCVANCEENSKIPQIILNMCEISKTYNPSRSDDILLLKELYSLLYALIEEFPKPFE
                     YKDKELHTYIQDALNFINSNYMHSITVQEIADYVNLSRSYLYKMFIKNLGISPQRYLIN
                     LRMYKATLLLKSTKLPIGEVASSVGYSDSLLFSKTFSKHFSMSPLNYRNNQVNKPSI"
     gene            complement(2584..3423)
                     /gene="bgaR"
                     /label="bgaR"
     protein_bind    3854..3870
                     /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)."
     CDS             3889..4221
                     /gene="mazF"
                     /label="Endoribonuclease toxin MazF"
                     /note="Endoribonuclease toxin MazF from Escherichia coli
                     (strain K12). Accession#: P0AE70"
     CDS             complement(4364..5095)
                     /gene="ermC'"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from Bacillus
                     subtilis. Accession#: P13956"
     primer_bind     complement(5627..5643)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5651..5667)
                     /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(5675..5705)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5720..5741)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6029..6617)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    7006..7130
                     /label="attR1"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        7155..7185
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"