LOCUS       40924_13545        5079 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCS2+gli3GFP, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5079)
  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  JOURNAL   Unpublished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 5079)
  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 5079)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5079)
  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            "Unpublished"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..5079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             257..973
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        complement(1027..1045)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    complement(1050..1184)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(1297..1315)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1336..1352)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1360..1376)
                     /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(1384..1414)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1429..1450)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1738..2326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2500..3357)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3358..3462)
                     /label=AmpR promoter
     rep_origin      complement(3488..3943)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4017..5001
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus
                     major immediate early transcription unit IE94"
     promoter        5036..5054
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 ggatcctatg gaagcccagt cccgtagcac tacagcaagt gaaaagaaaa aggttgagaa
       61 ctctatagta aaaggtcact ctcgaacaga agtcagtgag aaagctgttg cctcaagcac
      121 aacttccaat gaggatgaaa gtcctggaca gacatatcac agagaaagaa gaaacgccat
      181 tgccatgcag cctcaaggag gccaaggtct tggtaaaatc agcgaagagc cctctacatc
      241 aagtgaggaa agggccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      301 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      361 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
      421 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
      481 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
      541 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
      601 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
      661 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
      721 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
      781 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
      841 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
      901 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
      961 cgagctgtac aagggcggtg gaagatctgg gaattcaagg cctctcgagc ctctagattc
     1021 tgcagcccta tagtgagtcg tattacgtag atccagacat gataagatac attgatgagt
     1081 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     1141 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     1201 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaattc gcggccgcgg
     1261 cgccaatgca ttgggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg
     1321 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     1381 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     1441 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     1501 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     1561 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1621 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1681 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1741 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1801 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1861 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1921 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1981 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2041 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2101 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2161 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2221 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2281 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2341 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2401 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2461 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2521 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2581 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2641 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2701 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2761 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2821 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2881 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     2941 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3001 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3061 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3121 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3181 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3241 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3301 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3361 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3421 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3481 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     3541 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     3601 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     3661 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     3721 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     3781 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     3841 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     3901 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg
     3961 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagtcgacc
     4021 atagccaatt caatatggcg tatatggact catgccaatt caatatggtg gatctggacc
     4081 tgtgccaatt caatatggcg tatatggact cgtgccaatt caatatggtg gatctggacc
     4141 ccagccaatt caatatggcg gacttggcac catgccaatt caatatggcg gacttggcac
     4201 tgtgccaact ggggaggggt ctacttggca cggtgccaag tttgaggagg ggtcttggcc
     4261 ctgtgccaag tccgccatat tgaattggca tggtgccaat aatggcggcc atattggcta
     4321 tatgccagga tcaatatata ggcaatatcc aatatggccc tatgccaata tggctattgg
     4381 ccaggttcaa tactatgtat tggccctatg ccatatagta ttccatatat gggttttcct
     4441 attgacgtag atagcccctc ccaatgggcg gtcccatata ccatatatgg ggcttcctaa
     4501 taccgcccat agccactccc ccattgacgt caatggtctc tatatatggt ctttcctatt
     4561 gacgtcatat gggcggtcct attgacgtat atggcgcctc ccccattgac gtcaattacg
     4621 gtaaatggcc cgcctggctc aatgcccatt gacgtcaata ggaccaccca ccattgacgt
     4681 caatgggatg gctcattgcc cattcatatc cgttctcacg ccccctattg acgtcaatga
     4741 cggtaaatgg cccacttggc agtacatcaa tatctattaa tagtaacttg gcaagtacat
     4801 tactattgga aggacgccag ggtacattgg cagtactccc attgacgtca atggcggtaa
     4861 atggcccgcg atggctgcca agtacatccc cattgacgtc aatggggagg ggcaatgacg
     4921 caaatgggcg ttccattgac gtaaatgggc ggtaggcgtg cctaatggga ggtctatata
     4981 agcaatgctc gtttagggaa ccgccattct gcctggggac gtcggagcaa gcttgattta
     5041 ggtgacacta tagaatacaa gctacttgtt ctttttgca
//
