EN.CFP.SV40 vector (V009997)

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)."