LOCUS       sequence-258410-        6177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  enhancer reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6177)
  AUTHORS   Williams RM, Candido-Ferreira I, Repapi E, Gavriouchkina D, 
            Senanayake U, Ling ITC, Telenius J, Taylor S, Hughes J, 
            Sauka-Spengler T
  TITLE     Reconstruction of the Global Neural Crest Gene Regulatory Network In
            Vivo.
  JOURNAL   Dev Cell. 2019 Oct 21;51(2):255-276.e7. doi: 
            10.1016/j.devcel.2019.10.003.
  PUBMED    31639368
REFERENCE   2  (bases 1 to 6177)
  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"; doi: 
            "10.1016/j.devcel.2019.10"; journalName: "Dev Cell"; date: 
            "2019-10-21- 21"; volume: "51"; issue: "2"; pages: "255-276.e7"
COMMENT     Sequence Label: pTK_Sox10_enh_89_Citrine
FEATURES             Location/Qualifiers
     source          1..6177
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..92
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     41..60
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        1936..2081
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2159..2291
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2274..2293
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2345..2354
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2351..2353,2354..2356,2357..3067)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2396..2417)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2657..2676)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3004..3025
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3239..3360
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3250..3269)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3304..3323
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3608..3625)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3779..4367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3859..3878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4538..5329,5330..5398))
                     /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     5161..5180
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5399..5503)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5530..5985
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5617..5636)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5827..5848
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6116..6164
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
       61 cagtgcaagt gcaggtgcca gaacatttct ctatcgatag gtaccgagct cgagttgcca
      121 gggtcagtag gcagcactga ggggttacac tgctgagagg agaaaaaatt tcttgcaggg
      181 catccccatg cacaacagga tttacagcaa tggcttggac atcacaacct ctccaacaac
      241 aagtgttggc ttcagcttac aagatgcaaa atcaatagat gtagaggcaa gacagcgact
      301 tttctatctt tttaaactaa atgcagtgtt cctgtaaaaa tctagaccca ttgtagccac
      361 cactgaaccc ctgccagttc tgagtcccca tcatttgctt tttctgaaac aggcaccatg
      421 actccattcg taagacattt ccattagcat gtcgggctga ttcaacttat atcctgccct
      481 gttgtgtctg actcattttt ccccatctcc aggagaagcc tgaaggaaag cagagggccc
      541 ttcacataga tgctcagggt ctctatcaga taataactga agaaatattt gccatgaccg
      601 tgctatggtc attttttgcc attccttagt cacttggcag ccaatggagg gaagaggcga
      661 gagcagggtg ggggagagta aatgagacag agggacaaga acaaaagatt tctttgtagt
      721 gcagagagag agaaaaggga aagcatcacc aaatgcccag aagccatctt tgtaacaaga
      781 gacctattct ctctgcttct tggaaaacaa gttggagtaa aaggccctgg ttgaattaac
      841 ccttcacaga ggcaactcac tgagcatgaa ggaaagaggc atggccatga gaggcgtgat
      901 ctgtgtggtg aggtagtggc actggccctt aagcaattta aagcaaggat tggttgtctg
      961 ggaagaggag gtggggatga ggttgggtgg tttgaccata ccacagatct aaaatgtcag
     1021 atgtcctttc ttcacagacg ctttggtttt ggcaaagacc ttcagagggg tttttgcagc
     1081 atatgaggca acaaatcttt tccatggaca tttttttctt gggaggcaga gagaaaggaa
     1141 aacattcccc ctcaaaatac acattttact gcaacataat cttgtccaag aaacctctgt
     1201 aaatttacct cctgtcactg cccttcaaaa attccattct tgcgcactac atctctctta
     1261 ttcatgtctt agaccttccc ttggctgagc agaacaattt tattggagtt aggggttccc
     1321 tatagaaatg gtaaacaaaa ggtaaacaaa atcctccatc cccaaaggag caaccactgt
     1381 ttttcatcat gaacactctc tcttagttct cttcccaccc tctctaagaa agcttcctct
     1441 tttactctgt ggacactttt gagtgaaatg agacttcagc aacgataata caagaaaaaa
     1501 gatatagcca tggaaagaca cgttgaacct atttggagat caaacgtggg atatcagtat
     1561 atgcgagatc attttgtatc tgagcttgct ccaagctctc tgctaattaa aagaagtgag
     1621 acagccaggt cagaaataaa ggctttttat ctccatgaaa atacagaaag aacagcaccc
     1681 tgcttctgga gcctcatccc acagagccaa gcaaagagaa gcaaaggata gattggaaat
     1741 gaatacagaa cgggagcagt gtccagccat gggtcagcca tccccctggg cgctgatggg
     1801 gtagatacct gttggctagc ccgggctcga gatctgcgat ctaagtaagc ttgctgcttc
     1861 atccccgtgg cccgttgctc gcgtttgctg gcggtgtccc cggaagaaat atatttgcat
     1921 gtctttagtt ctatgatgac acaaaccccg cccagcgtct tgtcattggc gaattcgaac
     1981 acgcagatgc agtcggggcg gcgcggtccc aggtccactt cgcatattaa ggtgacgcgt
     2041 gtggcctcga acaccgagcg accctgcagc gacccgctta acagcgtcaa cagcgtgccg
     2101 cagatcccaa gcttgcggcc gcttaagctg cagaagttgg tcgtgaggca ctgggcaggt
     2161 aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca
     2221 gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt
     2281 tctctccaca ggtgtccact cccaggttca atacagctct taagcggccg caagctagct
     2341 tgccgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     2401 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     2461 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     2521 ctggcccacc ctcgtgacca ccttcggcta cggcctgatg tgcttcgccc gctaccccga
     2581 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     2641 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2701 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2761 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     2821 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     2881 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2941 cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga
     3001 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     3061 gtacaagtga gcctctagac tccctcagcg gcgcgcctgg tgttcctcga gatcggagtc
     3121 ctaaatcgac tgccttagcg gtaggacagt agatcagtca gcgacgtgac gaaccctcgc
     3181 aggatcaaga tcccacgtgt ctagagtcgg ggcggccggc cgcttcgagc agacatgata
     3241 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     3301 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     3361 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     3421 taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccgtc gaccgatgcc
     3481 cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc
     3541 cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt
     3601 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3661 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3721 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3781 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3841 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3901 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3961 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4021 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4081 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4141 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4201 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4261 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4321 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4381 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4441 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4501 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4561 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4621 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4681 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4741 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4801 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4861 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4921 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4981 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5041 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5101 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5161 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5221 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5281 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5341 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5401 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5461 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5521 tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     5581 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     5641 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5701 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     5761 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5821 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     5881 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     5941 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttgccat tcgccattca
     6001 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagccca
     6061 agctaccatg ataagtaagt aatattaagg tacgggaggt acttggagcg gccgcaataa
     6121 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagtact
//
