LOCUS       Exported                5996 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Gene trapping vector pDB790, complete sequence.
ACCESSION   KF921010
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5996)
  AUTHORS   Balciuniene J, Nagelberg D, Walsh KT, 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, 619 (2013)
  PUBMED    24034702
REFERENCE   2  (bases 1 to 5996)
  AUTHORS   Balciuniene J, Balciunas D.
  TITLE     Gene trapping using gal4 in zebrafish
  JOURNAL   J Vis Exp 79, E50113 (2013)
  PUBMED    24121167
REFERENCE   3  (bases 1 to 5996)
  AUTHORS   Balciunas D.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-NOV-2013) Department of Biology, Temple University, 
            1900 N 12th St, Philadelphia, PA 19122, USA
REFERENCE   4  (bases 1 to 5996)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5996
                     /organism="Gene trapping vector pDB790"
                     /lab_host="Danio rerio"
                     /mol_type="other DNA"
                     /label=pTol2/14XUAS:mRFP, crygc:GFP
                     /note="pTol2/14XUAS:mRFP, crygc:GFP"
                     /db_xref="taxon:1458412"
     misc_feature    complement(1..202)
                     /label=miniTol2 3
                     /note="miniTol2 3"
     misc_feature    210..414
                     /label=X. laevis crygc promoter
                     /note="X. laevis crygc promoter"
     misc_binding    210..414
                     /label=14X Gal4 UAS binding site
                     /bound_moiety="14X Gal4 UAS"
     CDS             438..1154
                     /codon_start=1
                     /product="GFP"
                     /label=GFP
                     /protein_id="AHI43152.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGYKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     polyA_signal    1198..1332
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      1202..1393
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 late polyA"
     CDS             1901..2578
                     /codon_start=1
                     /product="mRFP"
                     /label=mRFP
                     /protein_id="AHI43153.1"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     polyA_signal    2589..2723
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      2593..2784
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 late polyA"
     misc_feature    complement(2796..3056)
                     /label=miniTol2 5
                     /note="miniTol2 5"
     misc_feature    complement(3068..3095)
                     /label=T7
                     /note="T7"
     promoter        complement(3069..3087)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(3094..3111)
                     /label=M13F
                     /note="M13F"
     primer_bind     complement(3094..3110)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3182..3250
                     /label=partial LacZ alpha
                     /note="partial LacZ alpha"
     rep_origin      3251..3706
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      complement(3256..3696)
                     /direction=LEFT
                     /label=F1 ori
                     /note="F1 ori"
     promoter        3784..3888
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3889..4749
                     /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             4087..4749
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /protein_id="AHI43154.1"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     rep_origin      4844..5526
                     /label=ColE1 origin
                     /note="ColE1 origin"
     rep_origin      4920..5508
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5796..5817
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5832..5862
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     misc_feature    5866..5888
                     /label=LacO
                     /note="LacO"
     protein_bind    5870..5886
                     /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)."
     misc_feature    5894..5914
                     /label=M13R
                     /note="M13R"
     primer_bind     5894..5910
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5920..5945
                     /label=SP6
                     /note="SP6"
     promoter        5928..5946
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"