Basic Vector Information
Alpha3000-MT-gli2-6xHIS 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 ℃.
Alpha3000-MT-gli2-6xHIS vector Sequence
LOCUS 40924_240 11145 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector Alpha3000-MT-gli2-6xHIS, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11145)
AUTHORS Cerrizuela S, Vega-Lopez GA, Palacio MB, Aybar MJ.
TITLE Gli2 is required for the induction and migration of Xenopus laevis
neural crest
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 11145)
AUTHORS Cerrizuela S, Vega-Lopez GA, Palacio MB, Aybar MJ.
TITLE Direct Submission
JOURNAL Submitted (05-JUL-2017) Developmental Biology, Instituto Superior de
Investigaciones Biologicas (INSIBIO, CONICET), Chacabuco 461, San
Miguel de Tucuman, Tucuman T4000ILI, Argentina
REFERENCE 3 (bases 1 to 11145)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11145)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(05-JUL-2017) Developmental Biology, Instituto Superior de
Investigaciones Biologicas (INSIBIO, CONICET), Chacabuco 461, San
Miguel de Tucuman, Tucuman T4000ILI, Argentina"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11145
/mol_type="other DNA"
/organism="synthetic DNA construct"
sig_peptide 21..275
/gene="Gli2"
CDS 24..53
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 63..92
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 102..131
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 141..170
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 180..209
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 243..272
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 417..4367
/gene="gli4"
/label=gli4
/note="Zinc finger protein GLI4 from Xenopus laevis.
Accession#: Q91661"
CDS 4377..4394
/label=6xHis
/note="6xHis affinity tag"
promoter complement(4405..4422)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal complement(4427..4561)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter complement(4660..4678)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5108..5124)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 5132..5148
/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)."
promoter complement(5156..5186)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5201..5222)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5510..6098)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6272..7129)
/label=AmpR
/note="beta-lactamase"
promoter complement(7130..7234)
/label=AmpR promoter
primer_bind 7708..7724
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
regulatory 8046..11130
/regulatory_class="promoter"
regulatory 10953..10958
/regulatory_class="TATA_box"
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