pMK2010 vector (V004721)

LOCUS       40924_30950        4892 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMK2010, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4892)
  AUTHORS   House BL, Mortimer MW, Kahn ML.
  TITLE     New recombination methods for Sinorhizobium meliloti genetics
  JOURNAL   Appl. Environ. Microbiol. 70 (5), 2806-2815 (2004)
  PUBMED    15128536
REFERENCE   2  (bases 1 to 4892)
  AUTHORS   House BL, Kahn ML.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-SEP-2003) School of Molecular Biosciences, Washington 
            State University, Abelson Hall Room 406A, Pullman, WA 99164, USA
REFERENCE   3  (bases 1 to 4892)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4892)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2004"; volume: "70"; issue: "5"; pages:
            "2806-2815"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-SEP-2003) School of Molecular Biosciences, Washington State 
            University, Abelson Hall Room 406A, Pullman, WA 99164, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    29..260
                     /label=attP1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(659..961)
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(1306..1962)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(1963..2065)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     protein_bind    complement(2210..2441)
                     /label=attP2
                     /note="recombination site for the Gateway(R) BP reaction 
                     (pDONR(TM)221 version)"
     CDS             2565..3371
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      3517..4105
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     oriT            4348..4457
                     /label=oriT
                     /note="incP origin of transfer"
     terminator      complement(4662..4689)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(4781..4867)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"