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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

GBT-S86910 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900lox66FRT (minimal)Gal4-VP16FRT (minimal)lox71SV40 poly(A) signaleBFPT7 promoterM13 fwdf1 oriAmpR promoterAmpRoriCAP binding sitelac promoterlac operatorM13 revSP6 promoter

Plasmid Resuspension protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

GBT-S8 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

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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