pFloxin-TAP vector (V006228)

LOCUS       40924_20211        4032 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFloxin-TAP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4032)
  AUTHORS   Singla V, Hunkapiller J, Norman AR, Skarnes WC, Reiter JF.
  TITLE     The Floxin system: an efficient method for creating diverse targeted
            modifications in gene trap embryonic stem cell lines
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4032)
  AUTHORS   Singla V, Reiter JF, Skarnes WC.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-JUL-2008) Biochemistry, University of California, San 
            Francisco, 1550 4th Street, Rock Hall 446, San Francisco, CA 94158, 
            USA
REFERENCE   3  (bases 1 to 4032)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4032)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-JUL-2008) Biochemistry, University of California, San Francisco,
            1550 4th Street, Rock Hall 446, San Francisco, CA 94158, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..525
                     /label=b-actin
                     /note="b-actin"
                     /regulatory_class="promoter"
     regulatory      517..526
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(529..562)
                     /label=lox66
                     /note="Right element (RE) mutant of loxP (Araki et al.,
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     intron          563..828
                     /note="En2"
     misc_feature    831..856
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             857..874
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             902..922
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     CDS             929..994
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
     polyA_signal    1184..1318
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    1336..1383
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(1407..1423)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1431..1447)
                     /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(1455..1485)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1500..1521)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1809..2397)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2571..3428)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3429..3533)
                     /label=AmpR promoter
     primer_bind     4007..4023
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"