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pMSP3535 vector (V004456)

Basic Vector Information

Vector Name:
pMSP3535
Length:
8353 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Bryan EM, Bae T, Kleerebezem M, Dunny GM.

pMSP3535 vector Vector Map

pMSP35358353 bp400800120016002000240028003200360040004400480052005600600064006800720076008000ermBPRepDRepERepGNisRoriT7 promoterPnisA; nisin inducible promoter

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 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_32405        8353 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMSP3535, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8353)
  AUTHORS   Bryan EM, Bae T, Kleerebezem M, Dunny GM.
  TITLE     Improved vectors for nisin-controlled expression in gram-positive 
            bacteria
  JOURNAL   Plasmid 44 (2), 183-190 (2000)
  PUBMED    10964628
REFERENCE   2  (bases 1 to 8353)
  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   3  (bases 1 to 8353)
  AUTHORS   Staddon JH, Bryan EM, Manias DA, Bae T, Kleerebezem M, 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 8353)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 8353)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid"; 
            date: "2000"; volume: "44"; issue: "2"; pages: "183-190"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "J. 
            Bacteriol."; date: "2004"; volume: "186"; issue: "8"; pages: 
            "2393-2401"
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..8353
                     /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="AAP60030.1"
                     /translation="MNHDECKTYIKNSLLEIRKLANIYTLETFKKELEKRNIYLETKSD
                     KYFSSEGEDYIYKLIENNKIIYSISGKKLTYKGKKSFSKHAILKQLNEKANQVN"
     CDS             2388..3878
                     /codon_start=1
                     /product="RepE"
                     /label=RepE
                     /note="plasmid replication; pAMbeta1 plasmid replicon"
                     /protein_id="AAP60031.1"
                     /translation="MNIPFVVETVLHDGLLKYKFKNSKIRSITTKPGKSKGAIFAYRSK
                     SSMIGGRGVVLTSEEAIQENQDTFTHWTPNVYRYGTYADENRSYTKGHSENNLRQINTF
                     FIDFDIHTAKETISASDILTTAIDLGFMPTMIIKSDKGYQAYFVLETPVYVTSKSEFKS
                     VKAAKIISQNIREYFGKSLPVDLTCNHFGIARIPRTDNVEFFDPNYRYSFKEWQDWSFK
                     QTDNKGFTRSSLTVLSGTEGKKQVDEPWFNLLLHETKFSGEKGLIGRNNVMFTLSLAYF
                     SSGYSIETCEYNMFEFNNRLDQPLEEKEVIKIVRSAYSENYQGANREYITILCKAWVSS
                     DLTSKDLFVRQGWFKFKKKRSERQRVHLSEWKEDLMAYISEKSDVYKPYLVTTKKEIRE
                     VLGIPERTLDKLLKVLKANQEIFFKIKPGRNGGIQLASVKSLLLSIIKVKKEEKESYIK
                     ALTNSFDLEHTFIQETLNKLAERPKTDTQLDLFSYDTG"
     CDS             4228..4365
                     /codon_start=1
                     /product="RepG"
                     /label=RepG
                     /protein_id="AAP60032.1"
                     /translation="MEKYLKNPITWIGLVLVVTWFLTKSSEFLIFGVCVLLLVFASQSD
                     "
     CDS             4614..5252
                     /codon_start=1
                     /product="NisR"
                     /label=NisR
                     /note="response regulator"
                     /protein_id="AAP60033.1"
                     /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"
     regulatory      8045..8353
                     /note="PnisA; nisin inducible promoter"
                     /regulatory_class="promoter"

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