Basic Vector Information
- Vector Name:
- pMCentr2
- Antibiotic Resistance:
- Kanamycin
- Length:
- 2586 bp
- Type:
- Structural Genomics Vectors
- Replication origin:
- ori
- Source/Author:
- Midwest Center for Structural Genomics
- Copy Number:
- High copy number
- 5' Primer:
- M13 fwd
- 3' Primer:
- M13 rev
pMCentr2 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 ℃.
pMCentr2 vector Sequence
LOCUS 40924_29941 2586 bp DNA circular SYN 01-JAN-1980 DEFINITION Plasmid with a kanamycin resistance marker and a TEV cleavage site, for creating a Gateway(R) entry vector by ligation-independent cloning (LIC). ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2586) AUTHORS Midwest Center for Structural Genomics TITLE Direct Submission REFERENCE 2 (bases 1 to 2586) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" COMMENT For LIC, linearize with SspI and treat with T4 DNA polymerase plus dGTP. FEATURES Location/Qualifiers source 1..2586 /mol_type="other DNA" /organism="synthetic DNA construct" terminator complement(268..295) /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator complement(387..473) /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 537..553 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 569..668 /label=attL1 /note="recombination site for the Gateway(R) LR reaction" CDS 671..691 /codon_start=1 /label=TEV site /note="tobacco etch virus (TEV) protease recognition and cleavage site" /translation="ENLYFQS" protein_bind complement(711..810) /label=attL2 /note="recombination site for the Gateway(R) LR reaction" promoter complement(828..846) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(851..867) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" CDS 980..1786 /codon_start=1 /label=KanR /note="aminoglycoside phosphotransferase" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" rep_origin 1936..2524 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
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