Basic Vector Information
- Vector Name:
- pMCI002
- Length:
- 3990 bp
- Type:
- Cloning vector
- Replication origin:
- R6K γ ori
- Source/Author:
- Uhlich GA, Chen CY.
pMCI002 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 ℃.
pMCI002 vector Sequence
LOCUS 40924_30011 3990 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pMCI002, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3990)
AUTHORS Uhlich GA, Chen CY.
TITLE A cloning vector for creation of Escherichia colilacZ translational
fusions and generation of linear template for chromosomal
integration
JOURNAL Plasmid 67 (3), 259-263 (2012)
PUBMED 22197962
REFERENCE 2 (bases 1 to 3990)
AUTHORS Chen C-Y., Uhlich GA.
TITLE Direct Submission
JOURNAL Submitted (26-SEP-2011) Molecular Characterization of Foodborne
Pathogens RU, US Department of Agriculture, 600 E. Mermaid Lane,
Wyndmoor, PA 19038, USA
REFERENCE 3 (bases 1 to 3990)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3990)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid";
date: "2012"; volume: "67"; issue: "3"; pages: "259-263"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(26-SEP-2011) Molecular Characterization of Foodborne Pathogens RU,
US Department of Agriculture, 600 E. Mermaid Lane, Wyndmoor, PA
19038, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3990
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 1..21
/label=LacIF primer binding site
/note="LacIF primer binding site"
primer_bind 51..76
/note="SpecF primer binding site; primer with additional
sequence at the 5' end 'AAGGAATT'"
CDS 228..1007
/gene="ant1"
/label=ant1
/note="Spectinomycin 9-adenylyltransferase from
Staphylococcus aureus (strain N315). Accession#: P0A0D1"
primer_bind complement(1257..1294)
/label=SpecR primer binding site
/note="SpecR primer binding site"
misc_feature 1279..1302
/label=multiple cloning site
/note="multiple cloning site"
CDS 1298..1408
/codon_start=1
/product="lacZ"
/label=lacZ
/note="lacZ reading frame for translational fusion"
/protein_id="AFA52512.1"
/translation="DPVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR"
primer_bind complement(1346..1367)
/label=LacZR primer binding site
/note="LacZR primer binding site"
primer_bind complement(1389..1409)
/label=LacZR2 primer binding site
/note="LacZR2 primer binding site"
rep_origin complement(1418..1806)
/direction=LEFT
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
CDS complement(1836..2393)
/codon_start=1
/gene="traI"
/product="TraI"
/label=traI
/note="TraI DNA relaxase"
/protein_id="AFA52510.1"
/translation="MIAKHVPMRSIKKSDFAELVKYITDEQGKTERLGHVRVTNCEANT
LPAVMAEVMATQHGNTRSEADKTYHLLVSFRAGEKPDAETLRAIEDRICAGLGFAEHQR
VSAVHHDTDNLHIHIAINKIHPTRNTIHEPYRAYRALADLCATLERDYGLERDNHETRQ
RVSENRANDMERHAGVESLVGWI"
gene complement(1836..2393)
/gene="traI"
/label=traI
CDS complement(2431..2799)
/label=traJ
/note="oriT-recognizing protein"
oriT complement(2832..2941)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
CDS 3167..3563
/codon_start=1
/gene="traK"
/product="TraK"
/label=traK
/note="oriT binding protein"
/protein_id="AFA52508.1"
/translation="MPKSYTDELAEWVESRAAKKRRRDEAAVAFLAVRADVEAALASGY
ALVTIWEHMRETGKVKFSYETFRSHARRHIKAKPADVPAPQAKAAEPAPAPKTPEPRRP
KQGGKAEKPAPAAAPTGFTFNPTPDKKD"
gene 3167..3563
/gene="traK"
/label=traK
promoter 3670..3772
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
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