Basic Vector Information
- Vector Name:
- pMSP3535-FT
- Length:
- 8588 bp
- Type:
- Shuttle vector
- Replication origin:
- ori
- Source/Author:
- Staddon JH, Bryan EM, Manias DA, Dunny GM.
pMSP3535-FT 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 ℃.
pMSP3535-FT vector Sequence
LOCUS 40924_32395 8588 bp DNA circular SYN 18-DEC-2018 DEFINITION Shuttle vector pMSP3535-FT, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8588) AUTHORS Staddon JH, Bryan EM, Manias DA, Dunny GM. TITLE Conserved target for group II intron insertion in relaxase genes of conjugative elements of gram-positive bacteria JOURNAL J. Bacteriol. 186 (8), 2393-2401 (2004) PUBMED 15060042 REFERENCE 2 (bases 1 to 8588) AUTHORS Staddon JH, Bryan EM, Manias DA, Dunny GM. TITLE Nisin-inducible pCF10 target of Ll.ltrB JOURNAL Unpublished REFERENCE 3 (bases 1 to 8588) AUTHORS Staddon JH, Bryan EM, Manias DA, Dunny GM. TITLE Direct Submission JOURNAL Submitted (21-MAY-2003) Microbiology, University of Minnesota, MMC 196, 420 Delaware Street S.E., Minneapolis, MN 55455, USA REFERENCE 4 (bases 1 to 8588) TITLE Direct Submission REFERENCE 5 (bases 1 to 8588) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J. Bacteriol."; date: "2004"; volume: "186"; issue: "8"; pages: "2393-2401" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted (21-MAY-2003) Microbiology, University of Minnesota, MMC 196, 420 Delaware Street S.E., Minneapolis, MN 55455, USA" COMMENT SGRef: number: 4; type: "Journal Article" FEATURES Location/Qualifiers source 1..8588 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 332..1066 /gene="ermBP" /label=ermBP /note="rRNA adenine N-6-methyltransferase from Enterococcus faecalis. Accession#: P0A4D5" CDS 2046..2345 /codon_start=1 /product="RepD" /label=RepD /protein_id="AAQ75102.1" /translation="MNHDECKTYIKNSLLEIRKLANIYTLETFKKELEKRNIYLETKSD KYFSSEGEDYIYKLIENNKIIYSISGKKLTYKGKKSFSKHAILKQLNEKANQVN" CDS 2388..3878 /codon_start=1 /product="RepE" /label=RepE /note="plasmid replication; pAMbeta1 plasmid replicon" /protein_id="AAQ75103.1" /translation="MNIPFVVETVLHDGLLKYKFKNSKIRSITTKPGKSKGAIFAYRSK SSMIGGRGVVLTSEEAIQENQDTFTHWTPNVYRYGTYADENRSYTKGHSENNLRQINTF FIDFDIHTAKETISASDILTTAIDLGFMPTMIIKSDKGYQAYFVLETPVYVTSKSEFKS VKAAKIISQNIREYFGKSLPVDLTCNHFGIARIPRTDNVEFFDPNYRYSFKEWQDWSFK QTDNKGFTRSSLTVLSGTEGKKQVDEPWFNLLLHETKFSGEKGLIGRNNVMFTLSLAYF SSGYSIETCEYNMFEFNNRLDQPLEEKEVIKIVRSAYSENYQGANREYITILCKAWVSS DLTSKDLFVRQGWFKFKKKRSERQRVHLSEWKEDLMAYISEKSDVYKPYLVTTKKEIRE VLGIPERTLDKLLKVLKANQEIFFKIKPGRNGGIQLASVKSLLLSIIKVKKEEKESYIK ALTNSFDLEHTFIQETLNKLAERPKTDTQLDLFSYDTG" CDS 4228..4365 /codon_start=1 /product="RepG" /label=RepG /protein_id="AAQ75104.1" /translation="MEKYLKNPITWIGLVLVVTWFLTKSSEFLIFGVCVLLLVFASQSD " CDS 4614..5252 /codon_start=1 /product="NisR" /label=NisR /note="response regulator" /protein_id="AAQ75105.1" /translation="MKTALEMRNYEVATHQNISLPLDITDFQGFDLILLDIMMSNIEGT EICKRIRREISTPIIFVSAKDTEEDIINGLGIGGDDYITKPFSLKQLVAKVEANIKREE RNKHAVHVFSEIRRDLGPITFYLEERRVCVNGQTIPLTCREYDILELLSQRTSKVYTRE DIYDDVYDEYSNALFRSISEYIYQIRSKFAPYDINPIKTVRGLGYQWHG" CDS 5245..6585 /gene="nisK" /label=nisK /note="Nisin biosynthesis sensor protein NisK from Lactococcus lactis subsp. lactis. Accession#: P42707" rep_origin 7090..7678 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" promoter 7936..7954 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature complement(7976..8246) /note="contains nucleotides 578-308 from pcfG gene of pCF10; Ll.ltrB group II intron target; transcribed from PnisA in forward orientation" regulatory complement(8280..8588) /note="PnisA; nisin-inducible promoter" /regulatory_class="promoter"
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