LOCUS       V008149                 8457 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V008149
VERSION     V008149
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8457)
  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  JOURNAL   Unpublished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 8457)
  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 8457)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8457)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            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"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8457
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     sig_peptide     100..354
                     /gene="Gli2"
     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             496..4446
                     /gene="gli4"
                     /label="Zinc finger protein GLI4"
                     /note="Zinc finger protein GLI4 from Xenopus laevis.
                     Accession#: Q91661"
     CDS             4456..4473
                     /label="6xHis"
                     /note="6xHis affinity tag"
     promoter        complement(4484..4501)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    complement(4506..4640)
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        complement(4753..4771)
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4792..4808)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(4816..4832)
                     /label="lac operator"
                     /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(4840..4870)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4885..4906)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5194..5782)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5956..6813)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(6814..6918)
                     /label="AmpR promoter"
     rep_origin      complement(6944..7399)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        7473..8457
                     /label="CMV IE94 promoter"
                     /note="enhancer/promoter region of simian cytomegalovirus
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga tttaaagcta tggagcaaaa gctcatttct
      121 gaagaggact tgaatgaaat ggagcaaaag ctcatttctg aagaggactt gaatgaaatg
      181 gagcaaaagc tcatttctga agaggacttg aatgaaatgg agcaaaagct catttctgaa
      241 gaggacttga atgaaatgga gcaaaagctc atttctgaag aggacttgaa tgaaatggag
      301 agcttgggcg acctcaccat ggagcaaaag ctcatttctg aagaggactt gaattcaagg
      361 cctctcgacg tggagcatta cctgcgttct gtgcacaaca gccccaccct gtctatgatc
      421 tctgcagccc gtgggctcag cccagcagaa gtggcccatg agcacttaaa agaaaggggt
      481 atctatggtt tagcacctcc gccccctggt accaccccta cagaatactg ccaccaaatg
      541 gccttcctgg caagtcatgc aagtccatac ggggagctcc tggtgcagag cgcagcagct
      601 ggaaatacgt cgcaccttca cgattacctc acccccatgg atgtatcaag attctccagc
      661 ccccgagtta caccgagatt aagccggaaa cgggcccttt ccatctctcc tctgtccgat
      721 gccagcattg acctgcagac catgatccgc acctccccca actcgctggt tgcttacatc
      781 aacaactctc ggagcagctc agccgccagc gggtcctacg ggcacctttc tgcaggagca
      841 ataagcccgg cgtttagctt ccccatccat aaaccctgta gcctatcagc agctctgagc
      901 cagcagcgta gcctgagttc ctcatttgga cacacccctt tgctccaccc atcgccgacg
      961 tttgcatccc gccagcaggg agcactcaca tcagccaatc cagctcctcc cagcaacaac
     1021 agctccgccc ctgactccgt cctgaacaag gtgagcagtg agtctgcggt gagcagtaca
     1081 gtcaatcagg tgatacacaa acgcagcaaa gtgaagactg aagaagaggc cgattcagtg
     1141 cgattccccc agcctccgga ccacctgact gatctgaaag aggatttgga taaggatgag
     1201 tgcaagcagg agcctgaaca tatatacgag accaactgcc actgggacgg ctgcagcaag
     1261 gagtttgata cccaggatca gctcgtacat cacatcaata atgaccatat ccatggggag
     1321 aagaaggagt ttgtgtgtcg ctggcaggac tgctcccggg agcagaaacc attcaaggcg
     1381 cagtatatgc tggtggttca catgagaagg cacacggggg agaaaccaca taaatgcacc
     1441 tttgaaggtt gtttcaaggc ctattccagg ctggagaacc tgaagacgca cttgcgttca
     1501 cacaccgggg agaagccata tgtctgtgac cacgagggat gcaataaggc gttttccaat
     1561 gcgtctgatc gagccaaaca tcagaacaga acccattcca atgagaaacc gtacatatgt
     1621 aaggtcccag gctgcacaaa acgttacacg gaccccagct ctctcaggaa acacgttaaa
     1681 actgtacatg gacctgaggc ccatgtgacc aagaagcacc gcaatgacat catccaaaaa
     1741 ccttcactac ccaaggagaa cggggacaat gaagctagtg ccaaactcag cggtagggag
     1801 cactccgaca gtgtctctag agaccaggag cactgcttgc agaccagaac catcaaaaca
     1861 gaagataaca tgatgcatca gtccagcccc ggtgggcagt cgtcatgcag cagtgagcct
     1921 tccccatatg gcaacaccaa caacattgac agtggagtag atgtgagcct tgcatggcag
     1981 gggagcttgg gagacctgtt tggcttggag gagaccagcc cggtggtaga ctccacagta
     2041 tcatcctggc agcgcagtgg tcggcctgca actccggaaa cacagcgtat ccacagtgca
     2101 gagactggaa cagctgaaga aagagaaatt aaagacaatg aaagattcct gctcatttat
     2161 gaacccaatg ccacctgcca gaacacccgg ctcccgacca tcagtgcaaa tggtttcgat
     2221 gtaataggtg tccccagcag tgtgctaatc aaccccagag caatagaact gtccatgaat
     2281 gatgtgacca tgatgaacca attgaatgag cgccgtgacg ccaccagcag caccctgagc
     2341 tcagcataca ccagccgcag gtccgccggc atctctcctt acttctcgag tcgccgctca
     2401 gctgaaacct cacagtttgg aggccgcttg aataattcca gctctgcaga ttcttacgat
     2461 cccatttcca ccgatgcctc tagacgctct gctgaggcca gccagcacag tggacttcct
     2521 aatcttctca acctcactcc tgctcagcat tacagattaa aggccaagta tgctgcagca
     2581 actggtggtc caccacccac tcctcttccc aacatggata ggataggttt gagaaacaag
     2641 ctgtccctga tggatggagc agactttcct ttgcctccct tccggcaact tcctgttcca
     2701 agacgttgtg ctgatggtgg tggaaatgca ggactaaccc ccatgtatcc ccatgaaatt
     2761 ccggggaata attcaaggcg ggctgctgac ccagtcagaa gaactgcggg tattgatgac
     2821 aagccactcc caaggttcag ccgctttcat agcatgaata gtatgaatac attgcaccct
     2881 ccatccctat cggaaaggag aaatggaggc ctgcagcact acacctgctc tgatggaggc
     2941 cttcacaggc acgtctattc tccccgacca cccagcatta gtgaaaatgt tgctatggag
     3001 gccatatctt gtgatgcaga tgtccctggt ggagatgatg acctcatgct cccagatgat
     3061 gttgttcagt acatcagatc tcagaacaga gaagccccag agcaaaacct acagacggag
     3121 tacagcagtc cagcccggaa tctacaaagc aacacaaaat ctttccataa caatacacca
     3181 gaacaaccca gggcccctgg ggcttacttg agcaggaact ttcctgcttt ggctgaatgc
     3241 cttggccaga ctgctaacat gcaggataac aacatgcccg ttcagtggaa cgaagtcagt
     3301 tcgggtaccg ttgatgtcag tgacttaccc aagcagcagt ttgcagcagg caacttagca
     3361 gttgttcagc aaaaacagaa ctttgcccag tatcagagct tcaaccaagc acctatgcag
     3421 agggctcaca atattatggg tcaggggcaa gagtctgtac agaggaatat cagtgttaat
     3481 ggacagcggt ttaattattt gcaacaaaga caacagcaga tgagccagtg ccaaattgtc
     3541 agttctgact tcatccccca acagaggtac agccagagcc agtcaatgct gagttcaaga
     3601 gcaatgcaag aaggccagag ccagatttct ccatcctgta ataacatggt agagaggcct
     3661 ggggttcaca cacatgctgc tccaagcaac acactgaatc atcagcgcct tgctgtgcac
     3721 ggagcaccca cgcagggttt tgcaaataac ttctccgtta accaagacgg gctccaccca
     3781 cccaacgcct acacagtcca gcctcagaaa aatggattgg agccacaaca gaacactctt
     3841 ggaatgtctg ggcaagcgtt caaccatgga atgattcagc caagaccccc agctgcccca
     3901 caccccccta acagacccag gaatatacac ccggtacatc accctcctta catgaggagt
     3961 ccgcaccctg tgagtgaact gagcccaggc caacaaacag cagaggcaac accaaaacga
     4021 accagtgaaa acactgatcc aactcctaaa gacaataatt tgctttatta ttctggacaa
     4081 attcacatgt acgagccaaa tggaaacttc ggttctggca tagactgcac agtacggcaa
     4141 ctacccacca tgccgtctcc tggggcaaat caggtcacaa gcactgtaga ctcccaaggt
     4201 cttgagcacc cccctgttat agattttgac gccatgatgg atgatgggga tcactcgagc
     4261 cttatgtcag gaaccttgag ccctggcctc ctgcagagct tctcccaaac atcctcccgc
     4321 cttacaactc ccaggaattc cctgacgctc ccttccatcc ctgctgggat aaacaacatg
     4381 gcaataggag acatgagctc catgctcaca acattggcgg aggaaagcaa gtttcttgct
     4441 attatgcaac tgcagcatca tcaccatcac cattaatcta gaactatagt gagtcgtatt
     4501 acgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     4561 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     4621 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     4681 gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc
     4741 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     4801 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4861 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4921 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4981 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5041 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5101 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5161 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5221 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5281 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5341 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5401 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5461 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5521 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5581 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     5641 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5701 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5761 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5821 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5881 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5941 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6001 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6061 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6121 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6181 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6241 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6301 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6361 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6421 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6481 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6541 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6601 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6661 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6721 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6781 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6841 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6901 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     6961 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     7021 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     7081 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     7141 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     7201 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     7261 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     7321 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     7381 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     7441 tcggtgcggg cctcttcgct attacgccag tcgaccatag ccaattcaat atggcgtata
     7501 tggactcatg ccaattcaat atggtggatc tggacctgtg ccaattcaat atggcgtata
     7561 tggactcgtg ccaattcaat atggtggatc tggaccccag ccaattcaat atggcggact
     7621 tggcaccatg ccaattcaat atggcggact tggcactgtg ccaactgggg aggggtctac
     7681 ttggcacggt gccaagtttg aggaggggtc ttggccctgt gccaagtccg ccatattgaa
     7741 ttggcatggt gccaataatg gcggccatat tggctatatg ccaggatcaa tatataggca
     7801 atatccaata tggccctatg ccaatatggc tattggccag gttcaatact atgtattggc
     7861 cctatgccat atagtattcc atatatgggt tttcctattg acgtagatag cccctcccaa
     7921 tgggcggtcc catataccat atatggggct tcctaatacc gcccatagcc actcccccat
     7981 tgacgtcaat ggtctctata tatggtcttt cctattgacg tcatatgggc ggtcctattg
     8041 acgtatatgg cgcctccccc attgacgtca attacggtaa atggcccgcc tggctcaatg
     8101 cccattgacg tcaataggac cacccaccat tgacgtcaat gggatggctc attgcccatt
     8161 catatccgtt ctcacgcccc ctattgacgt caatgacggt aaatggccca cttggcagta
     8221 catcaatatc tattaatagt aacttggcaa gtacattact attggaagga cgccagggta
     8281 cattggcagt actcccattg acgtcaatgg cggtaaatgg cccgcgatgg ctgccaagta
     8341 catccccatt gacgtcaatg gggaggggca atgacgcaaa tgggcgttcc attgacgtaa
     8401 atgggcggta ggcgtgccta atgggaggtc tatataagca atgctcgttt agggaac
//
