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GBT-S8 vector (V009959#)

Basic Vector Information

Basic Vector Information
Vector Name GBT-S8 Antibiotic Resistance Ampicillin
Length 6910 bp Type Conditional gene trap vector
Source Grajevskaja V, Camerota D, Bellipanni G, Balciuniene J, Balciunas D.

GBT-S8 vector Vector Map

lox66 FRT (minimal) Gal4-VP16 FRT (minimal) lox71 SV40 poly(A) signal eBFP T7 promoter M13 fwd f1 ori AmpR promoter AmpR ori CAP binding site lac promoter lac operator M13 rev SP6 promoter

GBT-S8 vector Sequence

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LOCUS       Exported                6910 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Conditional gene trap vector GBT-S8, complete sequence.
ACCESSION   MH450096
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6910)
  AUTHORS   Grajevskaja V, Camerota D, Bellipanni G, Balciuniene J, Balciunas D.
  TITLE     Analysis of a conditional gene trap reveals that tbx5a is required 
            for heart regeneration in zebrafish
  JOURNAL   PLoS ONE 13 (6), e0197293 (2018)
  PUBMED    29933372
REFERENCE   2  (bases 1 to 6910)
  AUTHORS   Camerota D, Balciunas D.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JUN-2018) Department of Biology, Temple University, 
            1900 N 12th St, Philadelphia, PA 19122, USA
REFERENCE   3  (bases 1 to 6910)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6910
                     /organism="Conditional gene trap vector GBT-S8"
                     /mol_type="other DNA"
                     /db_xref="taxon:2250237"
     protein_bind    complement(220..253)
                     /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)."
     protein_bind    259..292
                     /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)"
     CDS             469..1119
                     /codon_start=1
                     /product="Gal4-VP16"
                     /label=Gal4-VP16
                     /note="AUG-less Gal4-VP16"
                     /protein_id="AWX64029.1"
                     /translation="KLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKR
                     SPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKD
                     AVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVSSRSTPGIQISRAAPPTD
                     VSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDM"
     CDS             469..906
                     /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"
     protein_bind    complement(2150..2183)
                     /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)"
     protein_bind    2190..2223
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     polyA_signal    2358..2492
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(2501..3220)
                     /codon_start=1
                     /product="eBFP"
                     /label=eBFP
                     /protein_id="AWX64030.1"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     mobile_element  complement(join(3710..3970,1..202))
                     /mobile_element_type="transposon:miniTol2"
     promoter        complement(3983..4001)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4008..4024)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4165..4620
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4698..4802
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4803..5663
                     /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"
     rep_origin      5834..6422
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6710..6731
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6746..6776
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6784..6800
                     /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     6808..6824
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6842..6860
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"

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