LOCUS       40924_40687        9093 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector PSM-123456_RC-Gli3-DT, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9093)
  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            "Unpublished"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..9093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     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             379..5085
                     /gene="gli3"
                     /label=gli3
                     /note="Zinc finger protein GLI3 from Xenopus laevis.
                     Accession#: Q91660"
     CDS             5092..5109
                     /label=6xHis
                     /note="6xHis affinity tag"
     promoter        complement(5120..5137)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    complement(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    complement(5452..5468)
                     /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(5476..5506)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5521..5542)
                     /label=CAP binding site
                     /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(6592..7449)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7450..7554)
                     /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"
     promoter        8109..9093
                     /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 cctctcgagc ctctatcgat ggatgctcaa tcaagaagca ctacagcaag tgaaaagaaa
      421 aaggttgaga actctatagt aaaaggtcac tctcgaacag aagtcagtga gaaagctgtt
      481 gcctcaagca caacttccaa tgaggatgaa agtcctggac agacatatca cagagaaaga
      541 agaaacgcca ttgccatgca gcctcaagga ggccaaggtc ttggtaaaat cagcgaagag
      601 ccctctacat caagtgagga aagggcctca ttaataaaaa aggaactcca tggatctata
      661 acacatcttc ctgaaccctc agttccctac cggggcactg tctttgccat ggatcctagg
      721 aatggttata tagatcctca ttaccgatcc gacctcgatc ttttcccggc ctttcatcct
      781 cctgtgccaa ttgatgcaag acatcatgag ggaagatacc attatgaacc aactcccatt
      841 cctccgctgc atgtgcctac tgcattagca agcagcccaa catacccaga tctcccattc
      901 ttccggatat ctcctcatag aaatccagcc agtgcatctg attccccatt cagccctcct
      961 catccgtaca tcagtcctta catggactac atccgatctc tccacagcag cccctcactg
     1021 tctatgatat cggctgctcg tggactcagc cctaccgatg tcccacatgc tggcattagc
     1081 cctgctgaat attaccacca aatggcactg ctggctggac agcgaagccc atatgctgat
     1141 atttttccat cccctgcaac tgctggtgct ggagctaatg cccttcatat ggaatatctt
     1201 catgccatgg agagctcacg gttttccagt cccagactat cagcacgacc aagtcgcaag
     1261 agaacgctgt ctatatcacc cctttctgat catagctttg atctccagac aatgattcgc
     1321 aactctccaa attctctcgt tactatcctt aacaactccc gcagcagctc atctgctagt
     1381 ggctcttatg ggcacctagc agccagtgcc atcagtccag ctttgaattt tgcataccag
     1441 cctaccccag tgtctcttca gcaaatgcat caacagatta tgagtcgaca gcacagcata
     1501 ggctcggcct ttggacacag ccctccactt cttcacccag caccaacatt tccttctcaa
     1561 agaaccattc ctggtatccc cagtgttttg aatccagtgc aggtcagcat tggcccatca
     1621 gaggctgcac agcagaacaa gcccaccagt gagtcagcag tcagcagcac tggggaccta
     1681 ctgcacaaca agcgctctaa ggtaaagcct gaagaagatc atccaagccc tggagcagtg
     1741 tgcattcagg atcagccaga tggaatgact ctggtgaaag aggagggggt caaagatgaa
     1801 agtaaacaag aggctgaagt ggtctatgag acaaactgcc attgggaagg ctgctcgcga
     1861 gagtttgaca cacaggaaca actagtgcat catatcaaca atgaccacat acatggagag
     1921 aaaaaggagt ttgtgtgtcg atggctggat tgttcaagag agcaaaagcc gttcaaggct
     1981 cagtatatgc tggtggtcca catgagaaga catacaggag aaaagccaca caaatgcact
     2041 tttgaaggtt gttcaaaagc ctactccaga ctagaaaact tgaaaactca cctgagatct
     2101 cacactgggg agaaaccata tgtctgtgag catgagggct gcaacaaagc cttttcaaat
     2161 gcatctgaca gagccaagca ccagaaccgc actcattcaa acgagaaacc ttatgtttgc
     2221 aagatcccag ggtgtacaaa acgttacact gaccctagct cactcagaaa gcatgtaaag
     2281 acagtgcatg gtccagaggc ccatgtgacc aagaagcaga gaggggacat acacccaagg
     2341 cctccaccac caagagaacc aggaagccat tcacagtcga gatcaccagg ccaacagact
     2401 cagggtatac atggagagca caaggacctc agtaacacta cctcaaagca tgaagaatgc
     2461 ctgcaagtaa gatcagtgaa gactgaaaaa ccaatgtcat ctcagccaag ccctggtggt
     2521 aagtcttcat gcagcaggca gcagtccccc atcagcaact ataccaacag tgggatcgag
     2581 cttaatctaa acagtggagg cagcattgga gacttcagtg ccctagatga aaccccaatc
     2641 atggactcta ccatttccac cgcaaccaca ggacttggtt tacacggaag gagggttatg
     2701 acaggaacca aatggatgga gcaaataaaa gttgaaaaaa tgaaacaagt caatgggatg
     2761 cttccaagac tgaacccagt tccacctcat agagcaccaa ctttaccacc aatcactgga
     2821 aatggcgtgc agctgaacag caacatcagt ttgggaggat tagcacctgt acttccaaac
     2881 agaaatgacc tttctagcat ggacattaca gtcttgaata tgcttaacag aagagatgcc
     2941 aataccagca caataagttc agcctatatg agcagccgca ggtcagctgg aatttcacct
     3001 tgtttttcca gtcgaggatc cgctgaggca tctcagtttg agggacgggt tcagaattta
     3061 agtgttgcag gctcctatga tccaatttct actgatgcct cgagacgttc agctgaaaca
     3121 agccagtgtg atgggctgcc aaccctcctt agcctaactc cagcacagca atacaggctg
     3181 aaagctaaat atgctgctgc gacaggtggg cccccaccaa caccactgcc taatatggaa
     3241 agaatgagcc taaaaaccag aatggctctc ttgggtgact caagagattg tagattatca
     3301 cctcttcctt cggtcaatgt cccacggaga tgtgctgatg gtggagtagg ctcttatgga
     3361 agaaggcatt tgttgccaaa tgaacttcaa gttaatggaa ttcgtagagc agcggatcct
     3421 gtaaaaatgg tctctgataa tgtttctaat actagagttg taaggttcaa cagtctaaac
     3481 aacgttattc ctccagatgt cccaccacct atggaaaggc gcaatttaag tcttcaaaac
     3541 tatacccgca atgtgtattc accatgtcct cctagcataa gtgaaaatgt tgccctggaa
     3601 gcgatgatta tggaagcaga aggcaatttt gatgacgagg atttactgcc agatgacatg
     3661 gttcagtatc ttaactctcg gaaccaaaga ctttatgaaa ccattgaaac agatgcatta
     3721 cagaataatg ctggagaaag aaatggaaac aactttgaac aatcccatgt gaccagtaat
     3781 gtaataaatg aacagttcca cagtagtgaa caaacagatc tcatagaaaa taaaaatgac
     3841 ctcccaattc agtggaatga agtaagctct ggcagtgcag aatattctcc atcaagattg
     3901 aagtatgggc agcgctttcc cacacagcag aatcaacctt ttggattata taataacatg
     3961 atggtccaac agcaaaatgt accaaagagc gggttgtccc agcaaagggg ctatcaacat
     4021 catacacaaa acaacccaca agccccccag caaaatttgg atctttacaa taacagcaat
     4081 gtgtggagtg gtcagttagg caggggcaat cagtacattg atgacataga aagatcatca
     4141 ctgggacatt ctgcaacagg aagtctttgc aattcagcag ctcgaggaca aaaacttaca
     4201 accaatggtt tgccaatgaa tactggtcaa caaaactttg gcccatccac acattatagt
     4261 gtgcaatcag ctaatagagc tgaagtagtc cagaatgaaa acatcataaa tcaggaattc
     4321 atgcaacaaa tgacagctga acacctgatc tatggtatgc atttgctggg ggtgactcga
     4381 tctaatcaga caacatcagg gcaaaatgga aacacaactg acggaactcg aagtttcctg
     4441 tcaactactc aaaatggtgg cgagcagcag ccaacccttg caaagaattt ccagtctttt
     4501 gtaaactcta gtggaaatag caggcagaat ttacacagaa acaacctgcc atatctgcaa
     4561 gagcaaatat atgacactaa ccaacacatt ttcaaagtga acagtattaa gatggaaatg
     4621 cagagtcacc cacaacagca ctgtgcaaac gcacaaaact tctccggtca gttgtatgac
     4681 caaacagcaa cataccctca acaaattatg aaatcaggct ctgaacccgg tatggaagca
     4741 aactgtcttt tacaagaaag taacaataca aactcatcca agcttctttc acctggggct
     4801 aaccaagtga ctagcacagt cgataacatt gataacagtt tggatggggt acaaattgat
     4861 tttgatgcta tcattgatga tggtgatcat gccagcctca tttctggagt tctaagtcca
     4921 agtatcatcc agaatctctc ccgtaattct tcacgcctta ccactcctcg agcatctctg
     4981 acattcccag ccatacctgt tagtacatct aacatggcta ttggggacat gagttctttg
     5041 ctgacctcac ttgcagaaga aagcaagttt cttgctatta tgcaactgca gcatcatcac
     5101 catcaccatt aatctagaac tatagtgagt cgtattacgt agatccagac atgataagat
     5161 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     5221 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     5281 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaat
     5341 tcgcggccgc ggcgccaatg cattgggccc ggtacccagc ttttgttccc tttagtgagg
     5401 gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5461 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5521 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5581 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5641 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5701 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5761 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5821 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5881 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5941 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6001 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6061 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6121 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6181 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6241 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     6301 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6361 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6421 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6481 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6541 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6601 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6661 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6721 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6781 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6841 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6901 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6961 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7021 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7081 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7141 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7201 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7261 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7321 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7381 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7441 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7501 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7561 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     7621 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     7681 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     7741 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     7801 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     7861 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     7921 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     7981 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     8041 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     8101 cgccagtcga ccatagccaa ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg
     8161 tggatctgga cctgtgccaa ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg
     8221 tggatctgga ccccagccaa ttcaatatgg cggacttggc accatgccaa ttcaatatgg
     8281 cggacttggc actgtgccaa ctggggaggg gtctacttgg cacggtgcca agtttgagga
     8341 ggggtcttgg ccctgtgcca agtccgccat attgaattgg catggtgcca ataatggcgg
     8401 ccatattggc tatatgccag gatcaatata taggcaatat ccaatatggc cctatgccaa
     8461 tatggctatt ggccaggttc aatactatgt attggcccta tgccatatag tattccatat
     8521 atgggttttc ctattgacgt agatagcccc tcccaatggg cggtcccata taccatatat
     8581 ggggcttcct aataccgccc atagccactc ccccattgac gtcaatggtc tctatatatg
     8641 gtctttccta ttgacgtcat atgggcggtc ctattgacgt atatggcgcc tcccccattg
     8701 acgtcaatta cggtaaatgg cccgcctggc tcaatgccca ttgacgtcaa taggaccacc
     8761 caccattgac gtcaatggga tggctcattg cccattcata tccgttctca cgccccctat
     8821 tgacgtcaat gacggtaaat ggcccacttg gcagtacatc aatatctatt aatagtaact
     8881 tggcaagtac attactattg gaaggacgcc agggtacatt ggcagtactc ccattgacgt
     8941 caatggcggt aaatggcccg cgatggctgc caagtacatc cccattgacg tcaatgggga
     9001 ggggcaatga cgcaaatggg cgttccattg acgtaaatgg gcggtaggcg tgcctaatgg
     9061 gaggtctata taagcaatgc tcgtttaggg aac
//
