Basic Vector Information
- Vector Name:
- pMR361-K
- Antibiotic Resistance:
- Kanamycin
- Length:
- 7836 bp
- Type:
- Cloning vector
- Replication origin:
- R6K γ ori
- Source/Author:
- Narayanan AM, Ramsey MM, Stacy A, Whiteley M.
pMR361-K 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 ℃.
pMR361-K vector Sequence
LOCUS 40924_32195 7836 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pMR361-K, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7836)
AUTHORS Narayanan AM, Ramsey MM, Stacy A, Whiteley M.
TITLE Genomic resources for an oral pathogen
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7836)
AUTHORS Ramsey MM.
TITLE Direct Submission
JOURNAL Submitted (13-MAR-2017) Department of Molecular Biosciences, The
University of Texas at Austin, 2506 Speedway, NMS 3.254, Austin, TX
78712, USA
REFERENCE 3 (bases 1 to 7836)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7836)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(13-MAR-2017) Department of Molecular Biosciences, The University of
Texas at Austin, 2506 Speedway, NMS 3.254, Austin, TX 78712, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7836
/mol_type="other DNA"
/organism="synthetic DNA construct"
oriT 35..144
/label=oriT
/note="incP origin of transfer"
CDS 177..545
/label=traJ
/note="oriT-recognizing protein"
CDS 604..1344
/codon_start=1
/product="TraI"
/label=TraI
/protein_id="ARI46004.1"
/translation="MRSIKKSDFAELVKYITDEQGKTERLGHVRVTNCEANTLPAVMAE
VMATQHGNTRSEADKTYHLLVSFRAGEKPDAETLRAIEDRICAGLGFAEHQRVSAVHHD
TDNLHIHIAINKIHPTRNTIHEPYRAYRALADLCATLERDYGLERDNHETRQRVSENRA
NDMERHAGVESLVGWILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFG
VGMVAGPVAGGLLGAISLLPRAFR"
promoter complement(1616..1720)
/label=AmpR promoter
rep_origin 1918..2506
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2794..2815
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2830..2860
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2868..2884
/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 2892..2908
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
CDS 2962..4008
/codon_start=1
/product="transposase"
/label=transposase
/protein_id="ARI46007.1"
/translation="MEKKEFRVLIKYCFLKGKNTVEAKTWLDNEFPDSAPGKSTIIDWY
AKFKRGEMSTEDGERSGRPKEVVTDENIKKIHKMILNDRKMKLIEIAEALKISKERVGH
IIHQYLDMRKLCAKWVPRELTFDQKQRRVDDSKRCLQLLTRNTPEFFRRYVTMDETWLH
HYTPESNRQSAEWTATGEPSPKRGKTQKSAGKVMASVFWDAHGIIFIDYLEKGKTINSD
YYMALLERLKVEIAAKRPHMKKKKVLFHQDNAPCHKSLRTMAKIHELGFELLPHPPYSP
DLAPSDFFLFSDLKRMLAGKKFGCNEEVIAETEAYFEAKPKEYYQNGIKKLEGRYNRCI
ALEGNYVE"
mobile_element 4020..4098
/mobile_element_type="transposon:mariner"
/label=ner
/note="repeat end"
promoter complement(4104..4122)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 4123..4139
/note="M13 rev; common sequencing primer, one of multiple
similar variants"
CDS complement(4208..4999)
/label=KanR
/note="aminoglycoside phosphotransferase"
promoter 5517..5535
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
mobile_element 5541..5640
/mobile_element_type="transposon:mariner"
/label=ner
/note="repeat end"
CDS 6012..6686
/codon_start=1
/product="spectinomycin resistance cassette"
/label=spectinomycin resistance cassette
/note="SpcR"
/protein_id="ARI46006.1"
/translation="MFGSGVESGLKPNSDLDFLVVVSEPLTDQSKEILIQKIRPISKKI
GDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYGEWLQELYEQGYIPQKELNSDLTIMLY
QAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMDSSEELIDNYQDDETNSILTLCRMILT
MDTGKIIPKDIAGNAVAESSPLEHRERILLAVRSYLGENIEWTNENVNLTINYLNNRLK
KL"
rep_origin complement(6871..7227)
/direction=LEFT
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
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