LOCUS       Exported                4032 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFloxin-TAP, complete sequence.
ACCESSION   EU916836
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4032
                     /organism="Cloning vector pFloxin-TAP"
                     /mol_type="other DNA"
                     /db_xref="taxon:554307"
     regulatory      1..525
                     /regulatory_class="promoter"
                     /note="b-actin"
     regulatory      517..526
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_recomb     529..562
                     /label=lox66 site
                     /note="lox66 site"
     protein_bind    529..562
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     intron          563..828
                     /note="En2"
     misc_feature    831..856
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    857..997
                     /note="tandem affinity purification (TAP) tag; FLAG, TEV, 
                     6x His"
     CDS             857..874
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             902..922
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             929..994
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      1077..1298
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40"
     polyA_signal    1184..1318
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    1336..1383
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_recomb     1337..1383
                     /label=FRT site
                     /note="FRT site"
     primer_bind     complement(1407..1423)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1431..1447
                     /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(1455..1485)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1500..1521
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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(2568..3428)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="AmpR"
                     /protein_id="ACG76135.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3429..3533)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4007..4023
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"