LOCUS       Exported                5274 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFloxin-IRES-eGFP, complete sequence.
ACCESSION   GQ357182
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5274)
  AUTHORS   Singla V, Hunkapiller J.
  TITLE     Efficient engineering of novel alleles at gene trap loci in mouse 
            embryonic stem cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5274)
  AUTHORS   Singla V, Hunkapiller J.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JUL-2009) Biochemistry, UCSF, 1550 4th Street, San 
            Francisco, CA 94158, USA
REFERENCE   3  (bases 1 to 5274)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5274
                     /organism="Cloning vector pFloxin-IRES-eGFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:664657"
     regulatory      1..525
                     /regulatory_class="promoter"
                     /note="b-actin"
     regulatory      520..529
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_recomb     532..565
                     /label=lox66
                     /note="lox66"
     protein_bind    532..565
                     /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          566..831
                     /note="En2"
     misc_feature    858..1453
                     /note="IRES; internal ribosome entry site"
     misc_feature    879..1451
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    902..1453
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     regulatory      1500..1509
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1506..2225
                     /codon_start=1
                     /product="eGFP"
                     /label=eGFP
                     /note="enhanced green fluorescent protein"
                     /protein_id="ACU30838.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      2319..2540
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40"
     polyA_signal    2426..2560
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    2578..2625
                     /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_feature    2579..2625
                     /label=FRT site
                     /note="FRT site"
     misc_feature    complement(2625..5274)
                     /label=ampicillin resistance gene cassette
                     /note="ampicillin resistance gene cassette"
     primer_bind     complement(2649..2665)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2673..2689
                     /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(2697..2727)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2742..2763
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3051..3639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3810..4670)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="ACU30839.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4671..4775)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5249..5265
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"