Basic Vector Information
- Vector Name:
- pMRKO1
- Length:
- 5881 bp
- Type:
- Suicide vector
- Replication origin:
- R6K γ ori
- Source/Author:
- Ramsey M, Rumbaugh K, Whiteley M.
- Promoter:
- lac
pMRKO1 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 ℃.
pMRKO1 vector Sequence
LOCUS 40924_32310 5881 bp DNA circular SYN 18-DEC-2018
DEFINITION Suicide vector pMRKO1, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5881)
AUTHORS Ramsey M, Rumbaugh K, Whiteley M.
TITLE Aggregatibacter actinomycetemcomitans suicide vector
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5881)
AUTHORS Ramsey M, Rumbaugh K, Whiteley M.
TITLE Direct Submission
JOURNAL Submitted (09-AUG-2010) Molecular Genetics and Microbiology,
University of Texas at Austin, 2506 Speedway Blvd, Austin, TX 78712,
USA
REFERENCE 3 (bases 1 to 5881)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5881)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(09-AUG-2010) Molecular Genetics and Microbiology, University of
Texas at Austin, 2506 Speedway Blvd, Austin, TX 78712, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5881
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 171..759
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 1047..1068
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1083..1113
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1121..1137
/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)."
primer_bind 1145..1161
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
CDS 1229..1936
/codon_start=1
/label=mCherry
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
CDS 2310..2984
/codon_start=1
/gene="SpcR"
/product="spectinomycin resistance protein"
/label=SpcR
/protein_id="ADV78249.1"
/translation="MFGSGVESGLKPNSDLDFLVVVSEPLTDQSKEILIQKIRPISKKI
GDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYGEWLQELYEQGYIPQKELNSDLTIMLY
QAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMDSSEELIDNYQDDETNSILTLCRMILT
MDTGKIIPKDIAGNAVAESSPLEHRERILLAVRSYLGENIEWTNENVNLTINYLNNRLK
KL"
gene 2310..2984
/gene="SpcR"
/label=SpcR
rep_origin complement(3169..3525)
/direction=LEFT
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
oriT 4169..4278
/label=oriT
/note="incP origin of transfer"
CDS 4311..4679
/codon_start=1
/label=traJ
/note="oriT-recognizing protein"
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
promoter complement(5750..5854)
/label=AmpR promoter
This page is informational only.