PSM-123456_RC-Gli3-DT vector (V003141)

Basic Vector Information

      • Vector Name:
      • PSM-123456_RC-Gli3-DT
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 9093 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Vega Lopez GA, Cerrizuela S, Aguero TH, Palacio MB, Tribulo C, Aybar MJ.

PSM-123456_RC-Gli3-DT vector Vector Map

PSM-123456_RC-Gli3-DT9093 bp40080012001600200024002800320036004000440048005200560060006400680072007600800084008800SP6 promotergli3T7 promoterSV40 poly(A) signalT3 promoterM13 revlac operatorlac promoterCAP binding siteoribeta-lactamaseAmpR promoterf1 oriSCMV IE94 promoter

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_RC-Gli3-DT vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                9093 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector PSM-123456_RC-Gli3-DT, complete sequence.
ACCESSION   KY435761
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
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
                     /organism="Cloning vector PSM-123456_RC-Gli3-DT"
                     /mol_type="other DNA"
                     /label=pCS2+
                     /note="pCS2+"
                     /db_xref="taxon:1969594"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             100..5112
                     /codon_start=1
                     /gene="gli3"
                     /product="PSM-GLI3"
                     /label=gli3
                     /protein_id="ARB02210.1"
                     /translation="MEQKLISEEDLNEMEQKLISEEDLNEMEQKLISEEDLNEMEQKLI
                     SEEDLNEMEQKLISEEDLNEMESLGDLTMEQKLISEEDLNSRPLEPLSMDAQSRSTTAS
                     EKKKVENSIVKGHSRTEVSEKAVASSTTSNEDESPGQTYHRERRNAIAMQPQGGQGLGK
                     ISEEPSTSSEERASLIKKELHGSITHLPEPSVPYRGTVFAMDPRNGYIDPHYRSDLDLF
                     PAFHPPVPIDARHHEGRYHYEPTPIPPLHVPTALASSPTYPDLPFFRISPHRNPASASD
                     SPFSPPHPYISPYMDYIRSLHSSPSLSMISAARGLSPTDVPHAGISPAEYYHQMALLAG
                     QRSPYADIFPSPATAGAGANALHMEYLHAMESSRFSSPRLSARPSRKRTLSISPLSDHS
                     FDLQTMIRNSPNSLVTILNNSRSSSSASGSYGHLAASAISPALNFAYQPTPVSLQQMHQ
                     QIMSRQHSIGSAFGHSPPLLHPAPTFPSQRTIPGIPSVLNPVQVSIGPSEAAQQNKPTS
                     ESAVSSTGDLLHNKRSKVKPEEDHPSPGAVCIQDQPDGMTLVKEEGVKDESKQEAEVVY
                     ETNCHWEGCSREFDTQEQLVHHINNDHIHGEKKEFVCRWLDCSREQKPFKAQYMLVVHM
                     RRHTGEKPHKCTFEGCSKAYSRLENLKTHLRSHTGEKPYVCEHEGCNKAFSNASDRAKH
                     QNRTHSNEKPYVCKIPGCTKRYTDPSSLRKHVKTVHGPEAHVTKKQRGDIHPRPPPPRE
                     PGSHSQSRSPGQQTQGIHGEHKDLSNTTSKHEECLQVRSVKTEKPMSSQPSPGGKSSCS
                     RQQSPISNYTNSGIELNLNSGGSIGDFSALDETPIMDSTISTATTGLGLHGRRVMTGTK
                     WMEQIKVEKMKQVNGMLPRLNPVPPHRAPTLPPITGNGVQLNSNISLGGLAPVLPNRND
                     LSSMDITVLNMLNRRDANTSTISSAYMSSRRSAGISPCFSSRGSAEASQFEGRVQNLSV
                     AGSYDPISTDASRRSAETSQCDGLPTLLSLTPAQQYRLKAKYAAATGGPPPTPLPNMER
                     MSLKTRMALLGDSRDCRLSPLPSVNVPRRCADGGVGSYGRRHLLPNELQVNGIRRAADP
                     VKMVSDNVSNTRVVRFNSLNNVIPPDVPPPMERRNLSLQNYTRNVYSPCPPSISENVAL
                     EAMIMEAEGNFDDEDLLPDDMVQYLNSRNQRLYETIETDALQNNAGERNGNNFEQSHVT
                     SNVINEQFHSSEQTDLIENKNDLPIQWNEVSSGSAEYSPSRLKYGQRFPTQQNQPFGLY
                     NNMMVQQQNVPKSGLSQQRGYQHHTQNNPQAPQQNLDLYNNSNVWSGQLGRGNQYIDDI
                     ERSSLGHSATGSLCNSAARGQKLTTNGLPMNTGQQNFGPSTHYSVQSANRAEVVQNENI
                     INQEFMQQMTAEHLIYGMHLLGVTRSNQTTSGQNGNTTDGTRSFLSTTQNGGEQQPTLA
                     KNFQSFVNSSGNSRQNLHRNNLPYLQEQIYDTNQHIFKVNSIKMEMQSHPQQHCANAQN
                     FSGQLYDQTATYPQQIMKSGSEPGMEANCLLQESNNTNSSKLLSPGANQVTSTVDNIDN
                     SLDGVQIDFDAIIDDGDHASLISGVLSPSIIQNLSRNSSRLTTPRASLTFPAIPVSTSN
                     MAIGDMSSLLTSLAEESKFLAIMQLQHHHHHH"
     gene            100..5112
                     /gene="gli3"
                     /label=gli3
     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             5092..5109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(5120..5137)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    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    5452..5468
                     /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(5476..5506)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5521..5542
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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(6589..7449)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="ARB02211.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7450..7554)
                     /gene="bla"
                     /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"
     regulatory      8107..9093
                     /regulatory_class="promoter"
                     /note="SCMV IE94 promoter"
     promoter        8109..9093
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"

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