Basic Vector Information
- Vector Name:
- pMK2030
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 6926 bp
- Type:
- Reporter vector
- Replication origin:
- R6K γ ori
- Source/Author:
- Humann JL, Schroeder BK, Mortimer MW, House BL, Yurgel SN, Maloney SC, Ward KL, Fallquist HM, Ziemkiewicz HT, Kahn ML.
- Promoter:
- lac UV5
pMK2030 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 ℃.
pMK2030 vector Sequence
LOCUS 40924_30965 6926 bp DNA circular SYN 18-DEC-2018
DEFINITION Reporter vector pMK2030, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6926)
AUTHORS Humann JL, Schroeder BK, Mortimer MW, House BL, Yurgel SN, Maloney
SC, Ward KL, Fallquist HM, Ziemkiewicz HT, Kahn ML.
TITLE Construction and expression of sugar kinase transcriptional gene
fusions by using the Sinorhizobium meliloti ORFeome
JOURNAL Appl. Environ. Microbiol. 74 (21), 6756-6765 (2008)
PUBMED 18791020
REFERENCE 2 (bases 1 to 6926)
AUTHORS Mortimer MW, Kahn ML, Humann JL.
TITLE Direct Submission
JOURNAL Submitted (16-JUN-2008) Institute of Biological Chemistry,
Washington State University, Pullman, WA 99164-6340, USA
REFERENCE 3 (bases 1 to 6926)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6926)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
Environ. Microbiol."; date: "2008"; volume: "74"; issue: "21";
pages: "6756-6765"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(16-JUN-2008) Institute of Biological Chemistry, Washington State
University, Pullman, WA 99164-6340, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6926
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(20..408)
/direction=LEFT
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
oriT 579..688
/label=oriT
/note="incP origin of transfer"
CDS 888..2075
/codon_start=1
/label=TcR
/note="tetracycline efflux protein"
/translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVLFIM
QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
protein_bind complement(2229..2276)
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
protein_bind 2317..2436
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 2466..2496
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 2550..3206
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 3551..3853
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(3897..4021)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
protein_bind 4062..4109
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS 4211..4924
/codon_start=1
/label=yeGFP
/note="yeast-enhanced green fluorescent protein"
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
primer_bind 4966..4982
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
CDS 5020..6828
/codon_start=1
/label=GUS
/note="beta-glucuronidase"
/translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
primer_bind complement(6897..6913)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
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