Basic Vector Information
GBT-S8 vector Vector Map
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
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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