LOCUS       Exported                9093 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector PSM-123456_Gli3-DT, complete sequence.
ACCESSION   KY435760
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  JOURNAL   Unpublished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_Gli3-DT"
                     /mol_type="other DNA"
                     /label=pCS2+
                     /note="pCS2+"
                     /db_xref="taxon:1969593"
     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
                     /note="Mutant form of gli3 gene"
                     /protein_id="ARB02208.1"
                     /translation="MEQKLISEEDLNEMEQKLISEEDLNEMEQKLISEEDLNEMEQKLI
                     SEEDLNEMEQKLISEEDLNEMESLGDLTMEQKLISEEDLNSRPLEPLDVEAQSRSTTAS
                     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
                     /note="Ampicillin resistance"
                     /protein_id="ARB02209.1"
                     /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"