LOCUS       sequence-9531-ex        5574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5574)
  AUTHORS   Antonellis A, Huynh JL, Lee-Lin SQ, Vinton RM, Renaud G, Loftus SK, 
            Elliot G, Wolfsberg TG, Green ED, McCallion AS, Pavan WJ
  TITLE     Identification of neural crest and glial enhancers at the mouse 
            Sox10 locus through transgenesis in zebrafish.
  JOURNAL   PLoS Genet. 2008  . 4(9):e1000174.
  PUBMED    18773071
REFERENCE   2  (bases 1 to 5574)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS Genet.
            2008  . 4(9):e1000174."
COMMENT     Sequence Label: pE1B-C-F/Sox10-MCS9
FEATURES             Location/Qualifiers
     source          1..5574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      105..114
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             111..1763
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(177..195)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1804..1925
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1815..1834)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1869..1888
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2173..2190)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2344..2932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2424..2443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3103..3894,3895..3963))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3726..3745
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3964..4068)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4095..4550
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4182..4201)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4392..4413
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4681..4729
                     /note="synthetic polyadenylation signal"
     misc_feature    4743..4834
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4783..4802
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     protein_bind    4873..4897
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(join(5564..5574,1..14))
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ttgtacaaag tggtgatggg ctcgagatct cagggatata aagctgggtt ggtgttgctt
       61 tgaatagttc atcttaagct tggcattccg gtactgttgg taaagccacc atggaagacg
      121 ccaaaaacat aaagaaaggc ccggcgccat tctatccgct ggaagatgga accgctggag
      181 agcaactgca taaggctatg aagagatacg ccctggttcc tggaacaatt gcttttacag
      241 atgcacatat cgaggtggac atcacttacg ctgagtactt cgaaatgtcc gttcggttgg
      301 cagaagctat gaaacgatat gggctgaata caaatcacag aatcgtcgta tgcagtgaaa
      361 actctcttca attctttatg ccggtgttgg gcgcgttatt tatcggagtt gcagttgcgc
      421 ccgcgaacga catttataat gaacgtgaat tgctcaacag tatgggcatt tcgcagccta
      481 ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa aaaaagctcc
      541 caatcatcca aaaaattatt atcatggatt ctaaaacgga ttaccaggga tttcagtcga
      601 tgtacacgtt cgtcacatct catctacctc ccggttttaa tgaatacgat tttgtgccag
      661 agtccttcga tagggacaag acaattgcac tgatcatgaa ctcctctgga tctactggtc
      721 tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt gagattctcg catgccagag
      781 atcctatttt tggcaatcaa atcattccgg atactgcgat tttaagtgtt gttccattcc
      841 atcacggttt tggaatgttt actacactcg gatatttgat atgtggattt cgagtcgtct
      901 taatgtatag atttgaagaa gagctgtttc tgaggagcct tcaggattac aagattcaaa
      961 gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa aagcactctg attgacaaat
     1021 acgatttatc taatttacac gaaattgctt ctggtggcgc tcccctctct aaggaagtcg
     1081 gggaagcggt tgccaagagg ttccatctgc caggtatcag gcaaggatat gggctcactg
     1141 agactacatc agctattctg attacacccg agggggatga taaaccgggc gcggtcggta
     1201 aagttgttcc attttttgaa gcgaaggttg tggatctgga taccgggaaa acgctgggcg
     1261 ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat tatgtccggt tatgtaaaca
     1321 atccggaagc gaccaacgcc ttgattgaca aggatggatg gctacattct ggagacatag
     1381 cttactggga cgaagacgaa cacttcttca tcgttgaccg cctgaagtct ctgattaagt
     1441 acaaaggcta tcaggtggct cccgctgaat tggaatccat cttgctccaa caccccaaca
     1501 tcttcgacgc aggtgtcgca ggtcttcccg acgatgacgc cggtgaactt cccgccgccg
     1561 ttgttgtttt ggagcacgga aagacgatga cggaaaaaga gatcgtggat tacgtcgcca
     1621 gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac gaagtaccga
     1681 aaggtcttac cggaaaactc gacgcaagaa aaatcagaga gatcctcata aaggccaaga
     1741 agggcggaaa gatcgccgtg taattctaga gtcggggcgg ccggccgctt cgagcagaca
     1801 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     1861 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     1921 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     1981 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat ccgtcgaccg
     2041 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc
     2101 gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
     2161 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2221 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2281 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2341 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2401 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2461 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2521 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2581 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2641 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2701 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2761 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2821 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2881 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2941 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3001 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3061 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3121 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3181 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3241 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3301 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3361 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3421 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3481 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3541 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3601 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3661 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3721 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3781 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3841 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3901 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3961 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4021 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4081 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4141 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4201 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4261 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4321 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4381 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4441 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4501 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt gccattcgcc
     4561 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     4621 gcccaagcta ccatgataag taagtaatat taaggtacgg gaggtacttg gagcggccgc
     4681 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta
     4741 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
     4801 cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat aggtaccgag ctcttacgcg
     4861 tgctagccca tcacaagttt gtacaaaaaa gcaggctgtg ctcactcaca gcactccgag
     4921 cctgtatgtc actcttgagg tcacagctgg aaagagcgcc caaggtcttc tggcgtttgc
     4981 cctaggctct ttccaaagtg gaccccaagc cagatagctc attcagaacg tctgggtgga
     5041 cagtggggtg ccaagcatca gcctaggcaa caacttctca ctggagtttg gaagagtaag
     5101 ggagagagaa gggaagaaag agggagtgaa gaagacaggg aggggatggg gggaggagga
     5161 agggaaagga gaggaggagg agagaggagg aggagggagg gaggggctgt ctgagaccca
     5221 gtcctcagca gctcaggaac aatcagacac tgtctctctc ctccaccccc tgggctgctg
     5281 ctggctttca tcactgacct cacccagagg gcagagcgct agggggaggc ggggggcagg
     5341 gggcagagga aggtcatagg tcagcctttg cgtgctgagc tggatcagga atagagctac
     5401 ttagggaatg gttgggggcg gggtgggggt gctcccgggc ccctcatgag gcacatccac
     5461 cccactcttg gtgttgtcag ctgatggaag gcaatggttg agaacagaca gttgccgtcc
     5521 actctaaaat ggtgggatct gtagctcctc ctttagccaa actacccagc tttc
//
