Basic Vector Information
- Vector Name:
- pMK2017
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 4252 bp
- Type:
- Cloning vector
- Replication origin:
- R6K γ ori
- Source/Author:
- House BL, Mortimer MW, Kahn ML.
- Promoter:
- lac UV5
pMK2017 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 ℃.
pMK2017 vector Sequence
LOCUS 40924_30960 4252 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pMK2017, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4252)
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 4252)
AUTHORS House BL, Mortimer MW, 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 4252)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4252)
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..4252
/mol_type="other DNA"
/organism="synthetic DNA construct"
oriT complement(93..202)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
rep_origin 373..761
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
misc_feature 772..877
/label=MCS
/note="pBluescript multiple cloning site"
protein_bind 898..945
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
protein_bind 1003..1122
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 1152..1182
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 1236..1892
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 2237..2539
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(2583..2707)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
protein_bind complement(2765..2812)
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS complement(2958..4145)
/codon_start=1
/label=TcR
/note="tetracycline efflux protein"
/translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVLFIM
QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
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