LOCUS       Exported                6888 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Gene trap vector pDB783, complete sequence.
ACCESSION   KF844243
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6888)
  AUTHORS   Balciuniene J, Nagelberg D, Walsh K, Camerota D, Georlette D, Biemar
            F, Bellipanni G, Balciunas D.
  TITLE     Efficient disruption of Zebrafish genes using a Gal4-containing gene
            trap
  JOURNAL   BMC Genomics 14 (1), 619 (2013)
  PUBMED    24034702
REFERENCE   2  (bases 1 to 6888)
  AUTHORS   Darius B.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-NOV-2013) Department of Biology, Temple University, 
            1900 N 12th St, Philadelphia, PA 19122, USA
REFERENCE   3  (bases 1 to 6888)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6888
                     /organism="Gene trap vector pDB783"
                     /mol_type="other DNA"
                     /db_xref="taxon:1441632"
     mobile_element  complement(2..203)
                     /mobile_element_type="transposon:Tol2"
                     /note="miniTol2 3"
     misc_feature    205..222
                     /label=I-Sce
                     /note="I-Sce"
     misc_feature    221..254
                     /label=loxP
                     /note="loxP"
     protein_bind    complement(221..254)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    263..402
                     /label=carp b-actin intron 1 splice acceptor
                     /note="carp b-actin intron 1 splice acceptor"
     CDS             423..1073
                     /codon_start=1
                     /product="AUG-less Gal4-VP16"
                     /label=AUG-less Gal4-VP16
                     /protein_id="AHE73662.1"
                     /translation="KLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKR
                     SPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKD
                     AVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVSSRSTPGIQISRAAPPTD
                     VSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDM"
     CDS             423..860
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="KLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKR
                     SPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKD
                     AVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     misc_feature    1080..1678
                     /label=Danio rerio b-actin 3 UTR
                     /note="Danio rerio b-actin 3 UTR"
     misc_feature    1659..2084
                     /label=Danio rerio b-actin p(A)
                     /note="Danio rerio b-actin p(A)"
     misc_feature    2089..2122
                     /label=FRT
                     /note="FRT"
     protein_bind    2089..2122
                     /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)"
     misc_feature    2131..2360
                     /label=SV40 poly(A)
                     /note="SV40 poly(A)"
     regulatory      2142..2333
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 late polyA"
     polyA_signal    2203..2337
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        2342..2359
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2379..3098)
                     /codon_start=1
                     /product="eGFP"
                     /label=eGFP
                     /protein_id="AHE73659.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      3095..3104
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    complement(3205..3574)
                     /label=14x Gal4 UAS
                     /note="14x Gal4 UAS"
     misc_feature    3588..3621
                     /label=FRT
                     /note="FRT"
     protein_bind    3588..3621
                     /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)"
     misc_feature    3632..3665
                     /label=loxP
                     /note="loxP"
     protein_bind    complement(3632..3665)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    complement(3666..3683)
                     /label=I-Sce
                     /note="I-Sce"
     mobile_element  complement(3689..3949)
                     /mobile_element_type="transposon:Tol2"
                     /note="miniTol2 5"
     primer_bind     complement(3961..3988)
                     /label=T7
                     /note="T7"
     promoter        complement(3962..3980)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3987..4004)
                     /label=M13F
                     /note="M13F"
     primer_bind     complement(3987..4003)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4075..4143
                     /codon_start=1
                     /product="LacZ alpha"
                     /label=LacZ alpha
                     /protein_id="AHE73660.1"
                     /translation="WRNSEEARTDRPSQQLRSLNGEW"
     rep_origin      4144..4599
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      complement(4149..4589)
                     /direction=LEFT
                     /label=F1 ori
                     /note="F1 ori"
     rep_origin      4161..4467
                     /label=F1 ori
                     /note="F1 ori"
     promoter        4677..4781
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4782..5642
                     /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"
     CDS             4980..5642
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="AHE73661.1"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     rep_origin      5737..6419
                     /label=ColE1 origin
                     /note="ColE1 origin"
     rep_origin      5813..6401
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6689..6710
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6725..6755
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     misc_feature    6759..6781
                     /label=LacO
                     /note="LacO"
     protein_bind    6763..6779
                     /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)."
     primer_bind     6787..6807
                     /label=M13R
                     /note="M13R"
     primer_bind     6787..6803
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6813..6838
                     /label=SP6
                     /note="SP6"
     promoter        6821..6839
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"