Basic Vector Information
PSM-123456_Gli3-DT 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 ℃.
PSM-123456_Gli3-DT vector Sequence
LOCUS 40924_40682 9093 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector PSM-123456_Gli3-DT, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9093)
AUTHORS Vega Lopez GA, Cerrizuela S, Aguero TH, Palacio MB, Tribulo C, Aybar
MJ.
TITLE The gli3 transcriptional activator is a key effector of Indian
hedgehog signaling during Xenopus neural crest development
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 9093)
AUTHORS Vega Lopez GA, Cerrizuela S, Aguero TH, Palacio MB, Tribulo C, Aybar
MJ.
TITLE Direct Submission
JOURNAL Submitted (03-JAN-2017) Developmental Biology, Instituto Superior de
Investigaciones Biologicas (INSIBIO, CONICET-UNT), Chacabuco 461,
Tucuman, Tucuman T4000ILI, Argentina
REFERENCE 3 (bases 1 to 9093)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9093)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(03-JAN-2017) Developmental Biology, Instituto Superior de
Investigaciones Biologicas (INSIBIO, CONICET-UNT), Chacabuco 461,
Tucuman, Tucuman T4000ILI, Argentina"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Assembly Method :: vector nti v. 11
Sequencing Technology :: Applied Biosystems 3130 Genetic Analyzer
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..9093
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 35..53
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
CDS 103..132
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 142..171
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 181..210
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 220..249
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 259..288
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 322..351
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 379..5085
/gene="gli3"
/label=gli3
/note="Zinc finger protein GLI3 from Xenopus laevis.
Accession#: Q91660"
CDS 5092..5109
/label=6xHis
/note="6xHis affinity tag"
promoter complement(5120..5137)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal complement(5142..5276)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter complement(5389..5407)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5428..5444)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5452..5468)
/label=lac operator
/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)."
promoter complement(5476..5506)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5521..5542)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5830..6418)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6592..7449)
/label=AmpR
/note="beta-lactamase"
promoter complement(7450..7554)
/label=AmpR promoter
rep_origin complement(7580..8035)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 8109..9093
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
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