p15TV-L vector (V009782)

Basic Vector Information

      • Vector Name:
      • p15TV-L
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 7746 bp
      • Type:
      • Expression vector
      • Source/Author:
      • Guthrie J, Loppnau P, Kozieradzki I, Savchenko A, Arrowsmith C.

p15TV-L vector Vector Map

p15TV-L7746 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600690072007500AmpR promoterAmpRoribomropCAP binding sitelacIlacI promoterT7 promoterlac operatorRBS6xHisTEV siteloxPsacB promoterSacB3'-LIC sequence; add complement to downstream PCR primerT7 terminatortet 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 ℃.

p15TV-L vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_2428        7746 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector p15TV-L, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7746)
  AUTHORS   Guthrie J, Loppnau P, Kozieradzki I, Savchenko A, Arrowsmith C.
  TITLE     Expression vectors for high-throughput In-Fusion cloning
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7746)
  AUTHORS   Guthrie J, Loppnau P, Kozieradzki I, Savchenko A, Arrowsmith C.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-FEB-2007) Structural Genomics Consortium, University 
            of Toronto, 100 College Street, Toronto, ON M5G1L5, Canada
REFERENCE   3  (bases 1 to 7746)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7746)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-FEB-2007) Structural Genomics Consortium, University of Toronto,
            100 College Street, Toronto, ON M5G1L5, Canada"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        104..208
                     /label=AmpR promoter
     CDS             209..1066
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1240..1828
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2014..2156)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2261..2449)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(3727..3748)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3764..4843)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(4844..4921)
                     /label=lacI promoter
     promoter        5230..5248
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5249..5273
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             5288..5310
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5329..5346
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             5359..5379
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     protein_bind    complement(5407..5440)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5455..5900
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             5901..7319
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     misc_feature    7395..7409
                     /note="3'-LIC sequence; add complement to downstream PCR
                     primer"
     terminator      7487..7534
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(7709..7737)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"

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