LOCUS       sequence-258448-        5482 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 5482)
  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 5482)
  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_84_Citrine
FEATURES             Location/Qualifiers
     source          1..5482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1148..1293
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1371..1503
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1486..1505
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1557..1566
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1563..1565,1566..1568,1569..2279)
                     /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(1608..1629)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1869..1888)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2216..2237
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2452..2573
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2463..2482)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2517..2536
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2821..2838)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2992..3580)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3072..3091)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3751..4542,4543..4611))
                     /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     4374..4393
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4612..4716)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4743..5198
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4830..4849)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5040..5061
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5329..5377
                     /note="synthetic polyadenylation signal"
     misc_feature    5391..5482
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5431..5450
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 atcgataggt accgagctcg agttgccagg tggccaagct tgctgggaaa gacaaaaccc
       61 atcccttctg atgaccttat gttttggtgc aggggataca atggtgctgg tgggctgagg
      121 catcttccct ctcatgaaat agatggggaa caagggacaa aagaactgga aaccagtccc
      181 caaattagag gctggtgtga gccctttgtg tatgggctca cagggcagag gggatgtaaa
      241 ggttggggtc ccaccactca aatgccggct ggtagctctg gttaccagcc caaatgcttc
      301 tgccctggct gtggtgcctc agcctggggc cgtgctctgt ggctcgctct ccctcacatt
      361 cctccgggag ctgatcaggg cttgaatgac accccagatg actccttgct attaattcct
      421 gcgaacagaa tgggctcctt gttctcccag tcctactctc tggaggaatt acaggggcca
      481 aggcagcggg ctgaagcctt cctgatttaa cccttccccc ggctccccag gagcttcccc
      541 gcttccactg ggttgttgga gctgaaagtc ttcccagagc tacctcggcc tccactgaac
      601 gagtccatga gggtcaccca gcactgttac aatcagagaa cccctccaga cagtatcctt
      661 ttctggtcat tctgcccaca tgtatttagg agtactcatt tcctttcttc tccccccggg
      721 catccttcct tggtcatgca gtcaaagaac gctcagttcc ctccattgta tcatgctgga
      781 catggaagat tccacctaag aaagcataaa actttctccc ccatctcctc attccttcag
      841 tacttgcttt ttgggatttg cttttttgga attacacctg gtctcttggt taggaggtga
      901 gggggttaat agtctgccac ccatcaaagc aggtagcaga cagacctgcc aggccttgct
      961 actctttgat ctttccaggc cctggtaatg cagggaattg tcccagggac aatgaggact
     1021 ctgttggcta gcccgggctc gagatctgcg atctaagtaa gcttgctgct tcatccccgt
     1081 ggcccgttgc tcgcgtttgc tggcggtgtc cccggaagaa atatatttgc atgtctttag
     1141 ttctatgatg acacaaaccc cgcccagcgt cttgtcattg gcgaattcga acacgcagat
     1201 gcagtcgggg cggcgcggtc ccaggtccac ttcgcatatt aaggtgacgc gtgtggcctc
     1261 gaacaccgag cgaccctgca gcgacccgct taacagcgtc aacagcgtgc cgcagatccc
     1321 aagcttgcgg ccgcttaagc tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca
     1381 aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac
     1441 tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca
     1501 caggtgtcca ctcccaggtt caatacagct cttaagcggc cgcaagctag cttgccgcca
     1561 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1621 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     1681 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     1741 ccctcgtgac caccttcggc tacggcctga tgtgcttcgc ccgctacccc gaccacatga
     1801 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     1861 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1921 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1981 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     2041 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     2101 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     2161 actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     2221 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     2281 gagcctctag actccctcag cggcgcgcct gtgtcgtagt tccaccgaaa gacggatagc
     2341 tgccagaact cgcttcggag catgaccaca gtgacccggt caaacgccct ctaaacagtc
     2401 agatctcacg tgtctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca
     2461 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2521 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2581 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     2641 agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccgat gcccttgaga
     2701 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt
     2761 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct cttccgcttc
     2821 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2881 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2941 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3001 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3061 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3121 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3181 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3241 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3301 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3361 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3421 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3481 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3541 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3601 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3661 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3721 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3781 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3841 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3901 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3961 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4021 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4081 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4141 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4201 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4261 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4321 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4381 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4441 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4501 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4561 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4621 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4681 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4741 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4801 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4861 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4921 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4981 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5041 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5101 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5161 taacgcgaat tttaacaaaa tattaacgct tacaatttgc cattcgccat tcaggctgcg
     5221 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc
     5281 atgataagta agtaatatta aggtacggga ggtacttgga gcggccgcaa taaaatatct
     5341 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct
     5401 ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt
     5461 gcaggtgcca gaacatttct ct
//
