EN.CFP.bGbpA vector (V009999)

Basic Vector Information

      • Vector Name:
      • EN.CFP.bGbpA
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 10298 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Chen SX, Osipovich AB, Ustione A, Potter LA, Hipkens S, Gangula R, Yuan W, Piston DW, Magnuson MA.

EN.CFP.bGbpA vector Vector Map

EN.CFP.bGbpA10298 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000T3 promoterM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoterf1 oriM13 fwdT7 promoterlox71Rosa 26 promoter5'UTRmCeruleanbeta-globin poly(A) signalFRT (minimal)PGK promoterEM7 promoterNeoR/KanRrabbit beta globin poly A signalFRT (minimal)lox2272

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.bGbpA vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_770       10298 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector EN.CFP.bGbpA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10298)
  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 10298)
  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 10298)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10298)
  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..10298
                     /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    complement(76..92)
                     /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)."
     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    7971..8026
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     protein_bind    8377..8410
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     promoter        8474..8973
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     promoter        8985..9032
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             9051..9851
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     regulatory      9868..10169
                     /label=rabbit beta globin poly A signal
                     /note="rabbit beta globin poly A signal"
                     /regulatory_class="polyA_signal_sequence"
     polyA_signal    9892..10116
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    10174..10207
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     protein_bind    10260..10293
                     /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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