Basic Vector Information
- Vector Name:
- Alpha-3000-gli3Act
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10139 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Vega Lopez GA, Cerrizuela S, Palacio MB, Aguero TH, Tribulo C, Aybar MJ.
Alpha-3000-gli3Act 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 ℃.
Alpha-3000-gli3Act vector Sequence
LOCUS 40924_235 10139 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector Alpha-3000-gli3Act, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10139)
AUTHORS Vega Lopez GA, Cerrizuela S, Palacio MB, Aguero TH, 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JOURNAL Unpublished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 10139)
AUTHORS Vega Lopez GA, Cerrizuela S, Palacio MB, Aguero TH, 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 10139)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10139)
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"Unpublished"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..10139
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 85..3606
/codon_start=1
/gene="Xlp-Sla-3000"
/product="gli3-Act"
/label=Xlp-Sla-3000
/note="Activator form of GLI3 transcription factor"
/protein_id="ARB02201.1"
/translation="MPGIPSVLNPVQVSIGPSEAAQQNKPTSESAVSSTGDLLHNKRSK
VKPEEDHPSPGAVCIQDQPDGMTLVKEEGVKDESKQEAEVVYETNCHWEGCSREFDTQE
QLVHHINNDHIHGEKKEFVCRWLDCSREQKPFKAQYMLVVHMRRHTGEKPHKCTFEGCS
KAYSRLENLKTHLRSHTGEKPYVCEHEGCNKAFSNASDRAKHQNRTHSNEKPYVCKIPG
CTKRYTDPSSLRKHVKTVHGPEAHVTKKQRGDIHPRPPPPREPGSHSQSRSPGQQTQGI
HGEHKDLSNTTSKHEECLQVRSVKTEKPMSSQPSPGGKSSCSRQQSPISNYTNSGIELN
LNSGGSIGDFSALDETPIMDSTISTATTGLGLHGRRVMTGTKWMEQIKVEKMKQVNGML
PRLNPVPPHRAPTLPPITGNGVQLNSNISLGGLAPVLPNRNDLSSMDITVLNMLNRRDS
NTSTISSAYMSSRRSSGISPCFSSRRSSEASQFEGRVQNLSVAGSYDPISTDASRRSSE
TSQCDGLPTLLSLTPAQQYRLKAKYAAATGGPPPTPLPNMERMSLKTRMALLGDSRDCR
LSPLPSVNVPRRCSDGGVGSYGRRHLLPNELQVNGMRRASDPVKMVSDNVSNTRVVRFN
SLNNVIPPDVPPPMERRNLSLQNYTRNVYSPCPPSISENVALEAMIMEAEGNFDDEDLL
PDDMVQYLNSRNQRLYETIETDALQNNAGERNGNNFEQSHVTSNVINEQFHSSEQTDLI
ENKNDLPIQWNEVSSGSAEYSPSRLKYGQRFPTQQNQPFGLYNNMMVQQQNVPKSGLSQ
QRGYQHHTQNNPQAPQQNLDLYNNSNVWSGQLGRGNQYIDDIERSSLGHSATGSLCNSA
ARGQKLTTNGLPMNTGQQNFGPSTHYSVQSANRAEVVQNENIINQEFMQQMTAEHLIYG
MHLLGVTRSNQTTSGQNGNTTDGTRSFLSTTQNGGEQQPTLAKNFQSFVNSSGNSRQNL
HRNNLPYLQEQIYDTNQHIFKVNSIKMEMQSHPQQHCANAQNFSGQLYDQTATYPQQIM
KSGSEPGMEANCLLQESNNTNSSKLLSPGANHVTSTVDNIDNSLAGVQIAFAAIIDDGD
HASLISGVLSPSIIQNLSRNSSRLTTPRASLTFPAIPVSTSNMAIGDMSSLLTSLAEES
KFLAIMQ"
promoter complement(3614..3631)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal complement(3636..3770)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4181..4197)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 4205..4221
/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(4229..4259)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4274..4295)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4583..5171)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5345..6202)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(6203..6307)
/label=AmpR promoter
primer_bind 6781..6797
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
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