LOCUS       Exported                7034 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFlipTrap, complete sequence.
ACCESSION   JN564735
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7034)
  AUTHORS   Trinh Le A, Hochgreb T, Graham M, Wu D, Ruf-Zamojski F, Jayasena CS,
            Saxena A, Hawk R, Gonzalez-Serricchio A, Dixson A, Chow E, Gonzales 
            C, Leung HY, Solomon I, Bronner-Fraser M, Megason SG, Fraser SE.
  TITLE     A versatile gene trap to visualize and interrogate the function of 
            the vertebrate proteome
  JOURNAL   Genes Dev. 25 (21), 2306-2320 (2011)
  PUBMED    22056673
REFERENCE   2  (bases 1 to 7034)
  AUTHORS   Trinh LA.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-AUG-2011) Biology, California Institute of Technology,
            1200 E. California Blvd, 139-74, Pasadena, CA 91125, USA
REFERENCE   3  (bases 1 to 7034)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7034
                     /organism="Cloning vector pFlipTrap"
                     /mol_type="other DNA"
                     /db_xref="taxon:1112871"
     misc_recomb     548..581
                     /label=FRT
                     /note="FRT"
     protein_bind    548..581
                     /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)"
     gene            644..2104
                     /gene="RalGDS/AF-6"
                     /label=RalGDS/AF-6
                     /note="Ras association domain family 8b"
     intron          644..969
                     /gene="RalGDS/AF-6"
                     /note="splice acceptor"
     exon            970..1779
                     /gene="citrine"
                     /note="synthetic exon"
     gene            970..1779
                     /gene="citrine"
                     /label=citrine
     misc_recomb     complement(1000..1033)
                     /label=loxP
                     /note="loxP"
     protein_bind    1000..1033
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             1039..1752
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     intron          1780..2104
                     /gene="RalGDS/AF-6"
                     /note="splice donor"
     misc_recomb     complement(2111..2144)
                     /label=loxPV
                     /note="loxPV"
     protein_bind    2111..2144
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     regulatory      complement(2151..2506)
                     /regulatory_class="polyA_signal_sequence"
                     /note="alpha tubulin"
     CDS             complement(2513..3220)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     exon            complement(2517..3225)
                     /gene="mCherry"
                     /note="3 exon"
     gene            complement(2517..3225)
                     /gene="mCherry"
                     /label=mCherry
     misc_recomb     3238..3271
                     /label=loxP
                     /note="loxP"
     protein_bind    complement(3238..3271)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_recomb     3278..3311
                     /label=loxPV
                     /note="loxPV"
     protein_bind    complement(3278..3311)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     misc_recomb     3318..3351
                     /label=FRT-F3
                     /note="FRT-F3"
     mobile_element  join(3357..3878,1..541)
                     /mobile_element_type="transposon:Tol2"
     primer_bind     complement(3912..3928)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3965..3983)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4004..4020)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4028..4044
                     /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(4052..4082)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4097..4118
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4406..4994)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5165..6025)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6026..6130)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6156..6611)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     6753..6769
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6779..6797
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"