Basic Vector Information
- Vector Name:
- pMK2010
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 4892 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- House BL, Mortimer MW, Kahn ML.
pMK2010 vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pMK2010 vector Sequence
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"
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