Basic Vector Information
- Vector Name:
- pMSP3535H3
- Length:
- 9463 bp
- Type:
- Shuttle vector
- Replication origin:
- ori
- Source/Author:
- Oddone GM, Mills DA, Block DE.
pMSP3535H3 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 ℃.
pMSP3535H3 vector Sequence
LOCUS 40924_32410 9463 bp DNA circular SYN 18-DEC-2018
DEFINITION Shuttle vector pMSP3535H3, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9463)
AUTHORS Oddone GM, Mills DA, Block DE.
TITLE Incorporation of nisI-mediated nisin immunity improves vector-based
nisin-controlled gene expression in lactic acid bacteria
JOURNAL Plasmid 61 (3), 151-158 (2009)
PUBMED 19141301
REFERENCE 2 (bases 1 to 9463)
AUTHORS Oddone GM, Kim J-H.
TITLE Direct Submission
JOURNAL Submitted (16-DEC-2008) Viticulture and Enology, University of
California, Davis, RMI North Lab, 595 Hilgard Lane, Davis, CA 95616,
USA
REFERENCE 3 (bases 1 to 9463)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9463)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid";
date: "2009"; volume: "61"; issue: "3"; pages: "151-158"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(16-DEC-2008) Viticulture and Enology, University of California,
Davis, RMI North Lab, 595 Hilgard Lane, Davis, CA 95616, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9463
/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 1971..2270
/codon_start=1
/gene="repD"
/product="RepD"
/label=repD
/note="replication factor"
/protein_id="ACM62692.1"
/translation="MNHDECKTYIKNSLLEIRKLANIYTLETFKKELEKRNIYLETKSD
KYFSSEGEDYIYKLIENNKIIYSISGKKLTYKGKKSFSKHAILKQLNEKANQVN"
gene 1971..2270
/gene="repD"
/label=repD
CDS 2313..3803
/codon_start=1
/gene="repE"
/product="RepE"
/label=repE
/note="replication factor"
/protein_id="ACM62693.1"
/translation="MNIPFVVETVLHDGLLKYKFKNSKIRSITTKPGKSKGAIFAYRSK
SSMIGGRGVVLTSEEAIQENQDTFTHWTPNVYRYGTYADENRSYTKGHSENNLRQINTF
FIDFDIHTAKETISASDILTTAIDLGFMPTMIIKSDKGYQAYFVLETPVYVTSKSEFKS
VKAAKIISQNIREYFGKSLPVDLTCNHFGIARIPRTDNVEFFDPNYRYSFKEWQDWSFK
QTDNKGFTRSSLTVLSGTEGKKQVDEPWFNLLLHETKFSGEKGLIGRNNVMFTLSLAYF
SSGYSIETCEYNMFEFNNRLDQPLEEKEVIKIVRSAYSENYQGANREYITILCKAWVSS
DLTSKDLFVRQGWFKFKKKRSERQRVHLSEWKEDLMAYISEKSDVYKPYLVTTKKEIRE
VLGIPERTLDKLLKVLKANQEIFFKIKPGRNGGIQLASVKSLLLSIIKVKKEEKESYIK
ALTNSFDLEHTFIQETLNKLAERPKTDTQLDLFSYDTG"
gene 2313..3803
/gene="repE"
/label=repE
CDS 4153..4290
/codon_start=1
/gene="repG"
/product="RepG"
/label=repG
/note="replication factor"
/protein_id="ACM62694.1"
/translation="MEKYLKNPITWIGLVLVVTWFLTKSSEFLIFGVCVLLLVFASQSD
"
gene 4153..4290
/gene="repG"
/label=repG
CDS 4440..5177
/codon_start=1
/gene="nisI"
/product="NisI"
/label=nisI
/note="nisin immunity protein; provides significant
protection from the toxic effects of the inducer nisin"
/protein_id="ACM62695.1"
/translation="MRKYLILIVALIGITGLSGCYQTSQKKVRFDEGSYTNFIYDNKSY
FVTDKEIPQENVNNCKVKFYNLLIVDMKSEKLLSSSNKNSVTLVLNNIYEASDKSLCMG
INDRYYKILPESDKGVVKALRLQNFDVTSDISDDNFVIDKNDSRKIDYMGNIYSISDTT
VSDEELGEYQDVLAEVRVFDSVSGKSIPRSEWGRIDKDGSNSKQSRTEWDYGEIHSIRG
KSLTEAFAVEINDDFKLATKVGN"
gene 4440..5177
/gene="nisI"
/label=nisI
CDS 5406..6044
/codon_start=1
/gene="nisR"
/product="NisR"
/label=nisR
/note="nis operon regulator protein"
/protein_id="ACM62696.1"
/translation="MKTALEMRNYEVATHQNISLPLDITDFQGFDLILLDIMMSNIEGT
EICKRIRREISTPIIFVSAKDTEEDIINGLGIGGDDYITKPFSLKQLVAKVEANIKREE
RNKHAVHVFSEIRRDLGPITFYLEERRVCVNGQTIPLTCREYDILELLSQRTSKVYTRE
DIYDDVYDEYSNALFRSISEYIYQIRSKFAPYDINPIKTVRGLGYQWHG"
gene 5406..6044
/gene="nisR"
/label=nisR
CDS 6037..7377
/gene="nisK"
/label=nisK
/note="Nisin biosynthesis sensor protein NisK from
Lactococcus lactis subsp. lactis. Accession#: P42707"
rep_origin 7882..8470
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 8728..8746
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 9147..9163
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 9164..9220
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(9233..9249)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
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