Basic Vector Information
Alpha-3000-gli2act 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-gli2act vector Sequence
LOCUS 40924_230 10802 bp DNA circular SYN 17-DEC-2018 DEFINITION Cloning vector Alpha-3000-gli2act, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10802) 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 TITLE Gli2 is required for the induction and migration of Xenopus laevis neural crest JOURNAL Unpublished REFERENCE 2 (bases 1 to 10802) AUTHORS Cerrizuela S, Vega-Lopez GA, Palacio MB, Aybar MJ.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 10802) TITLE Direct Submission REFERENCE 4 (bases 1 to 10802) 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 TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (05-JUL-2017) Developmental Biology, Instituto Superior de Investigaciones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..10802 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 6..4067 /codon_start=1 /gene="Gli2 activator" /product="Gli2act" /label=Gli2 activator /note="activator form of Gli2 protein, without the N-terminal repressor region" /protein_id="ATQ62667.1" /translation="MGGGALSISPLSDASIDLQTMIRTSPNSLVAYINNSRSSSAASGS YGHLSAGAISPAFSFPIHKPCSLSAALSQQRSLSSSFGHTPLLHPSPTFASRQQGALTS ANPAPPSNNSSAPDSVLNKVSSESAVSSTVNQVIHKRSKVKTEEEADSVRFPQPPDHLT DLKEDLDKDECKQEPEHIYETNCHWDGCSKEFDTQDQLVHHINNDHIHGEKKEFVCRWQ DCSREQKPFKAQYMLVVHMRRHTGEKPHKCTFEGCFKAYSRLENLKTHLRSHTGEKPYV CDHEGCNKAFSNASDRAKHQNRTHSNEKPYICKVPGCTKRYTDPSSLRKHVKTVHGPEA HVTKKHRNDIIQKPSLPKENGDNEASAKLSGREHSDSVSRDQEHCLQTRTIKTEDNMMH QSSPGGQSSCSSEPSPYGNTNNIDSGVDVSLAWQGSLGDLFGLEETSPVVDSTVSSWQR SGRPATPETQRIHSAETGTAEEREIKDNERFLLIYEPNATCQNTRLPTISANGFDVIGV PSSVLINPRAIELSMNDVTMMNQLNERRDSTSSTLSSAYTSRRSSGISPYFSSRRSSET SQFGGRLNNSSSADSYDPISTDASRRSSEASQHSGLPNLLNLTPAQHYRLKAKYAAATG GPPPTPLPNMDRIGLRNKLSLMDGADFPLPPFRQLPVPRRCSDGGGNAGLTPMYPHEIP GNNSRRASDPVRRTAGIDDKPLPRFSRFHSMNSMNTLHPPSLSERRNGGLQHYTCSDGG LHRHVYSPRPPSISENVAMEAISCDADVPGGDDDLMLPDDVVQYIRSQNREAPEQNLQT EYSSPARNLQSNTKSFHNNTPEQPRAPGAYLSRNFPALAECLGQTANMQDNNMPVQWNE VSSGTVDVSDLPKQQFAAGNLAVVQQKQNFAQYQSFNQAPMQRAHNIMGQGQESVQRNI SVNGQRFNYLQQRQQQMSQCQIVSSDFIPQQRYSQSQSMLSSRAMQEGQSQISPSCNNM VERPGVHTHAAPSNTLNHQRLAVHGAPTQGFANNFSVNQDGLHPPNAYTVQPQKNGLEP QQNTLGMSGQAFNHGMIQPRPPAAPHPPNRPRNIHPVHHPPYMRSPHPVSELSPGQQTA EATPKRTSENTDPTPKDNNLLYYSGQIHMYEPNGNFGSGIDCTVRQLPTMPSPGANQVT STVDSQGLEHPPVIDFDAMMDDGDHSSLMSGTLSPGLLQSFSQTSSRLTTPRNSLTLPS IPAGINNMAIGDMSSMLTTLAEESKFLNLNRFKAMEQKLISEEDLNEMEQKLISEEDLN EMEQKLISEEDLNEMEQKLISEEDLNEMEQKLISEEDLNEMESLGDLTMEQKLISEEDL NSRPLEPLEL" gene 6..4067 /gene="Gli2 activator" /label=Gli2 activator sig_peptide 3783..4037 /gene="Gli2 activator" CDS 3786..3815 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 3825..3854 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 3864..3893 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 3903..3932 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 3942..3971 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 4005..4034 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" promoter complement(4062..4079) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal complement(4084..4218) /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter complement(4317..4335) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4765..4781) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 4789..4805 /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(4813..4843) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(4858..4879) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(5167..5755) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(5929..6786) /label=AmpR /note="beta-lactamase" promoter complement(6787..6891) /label=AmpR promoter primer_bind 7365..7381 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" regulatory 7703..10787 /gene="Xlp-Sla-3000" /regulatory_class="promoter" gene 7703..10787 /gene="Xlp-Sla-3000" /label=Xlp-Sla-3000 regulatory 10610..10615 /gene="Xlp-Sla-3000" /regulatory_class="TATA_box"
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