Basic Vector Information
- Vector Name:
- EN.CFP.SV40
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9966 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Chen SX, Osipovich AB, Ustione A, Potter LA, Hipkens S, Gangula R, Yuan W, Piston DW, Magnuson MA.
- Promoter:
- mPGK
EN.CFP.SV40 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 ℃.
EN.CFP.SV40 vector Sequence
LOCUS 40924_775 9966 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector EN.CFP.SV40, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9966)
AUTHORS Chen SX, Osipovich AB, Ustione A, Potter LA, Hipkens S, Gangula R,
Yuan W, Piston DW, Magnuson MA.
TITLE Quantification of factors influencing fluorescent protein expression
using RMCE to generate an allelic series in the ROSA26 locus in mice
JOURNAL Dis Model Mech 4 (4), 537-547 (2011)
PUBMED 21324933
REFERENCE 2 (bases 1 to 9966)
AUTHORS Chen SX, Osipovich AB, Magnuson MA.
TITLE Direct Submission
JOURNAL Submitted (16-JUL-2010) Molecular Physiology and Biophysics,
Vanderbilt University, 9475 MRB IV, 2214 Garland Avenue, Nashville,
TN 37232, USA
REFERENCE 3 (bases 1 to 9966)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9966)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Dis Model
Mech"; date: "2011"; volume: "4"; issue: "4"; pages: "537-547"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(16-JUL-2010) Molecular Physiology and Biophysics, Vanderbilt
University, 9475 MRB IV, 2214 Garland Avenue, Nashville, TN 37232,
USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9966
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter complement(13..31)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(52..68)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 76..92
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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)."
promoter complement(100..130)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(145..166)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(454..1042)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1216..2073)
/label=AmpR
/note="beta-lactamase"
promoter complement(2074..2178)
/label=AmpR promoter
rep_origin complement(2253..2708)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 2850..2866
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 2873..2891
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 2906..2939
/label=lox71
/note="Left element (LE) mutant of loxP (Araki et al.,
2010). Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
regulatory 2957..7037
/label=Rosa 26 promoter
/note="Rosa 26 promoter"
/regulatory_class="promoter"
5'UTR 7050..7092
/label=Xenopus globin 5'-UTR
/note="translational enhancer from the 5'-UTR of the major
beta-globin gene of Xenopus laevis"
CDS 7103..7819
/label=mCerulean
/note="enhanced monomeric variant of CFP (Rizzo et al.,
2004)"
polyA_signal 7931..8012
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
protein_bind 8045..8078
/label=FRT (minimal)
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
promoter 8142..8641
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
promoter 8653..8700
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 8719..9519
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase from Tn5"
polyA_signal 9560..9784
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
misc_feature 9842..9875
/label=FRT site
/note="FRT site"
protein_bind 9842..9875
/label=FRT (minimal)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
protein_bind 9928..9961
/label=lox2272
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are
compatible with each other, but incompatible with loxP or
loxN sites (Lee and Saito, 1988)."
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