LOCUS       sequence-259315-        5458 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 5458)
  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 5458)
  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_Pax7_enh_194_Cerulean
FEATURES             Location/Qualifiers
     source          1..5458
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..49
                     /note="synthetic polyadenylation signal"
     misc_feature    63..154
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     103..122
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        1279..1424
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1502..1634
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1617..1636
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1688..1697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1694..1696,1697..1699,1700..2410)
                     /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(1739..1760)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2000..2019)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2347..2368
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2582..2703
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2593..2612)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2647..2666
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2951..2968)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3122..3710)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3202..3221)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3881..4672,4673..4741))
                     /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     4504..4523
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4742..4846)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4873..5328
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4960..4979)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5170..5191
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta
       61 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
      121 cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat aggtaccgag ctcgagttgc
      181 cagggctccg aaattcctcc ccaatccccc taccccaaat attttgagag ctgtaaaatg
      241 aggcggagat cgtcttaatt ctgtcggcag gacgagaatt gctttcgggt gcgtgcgtgt
      301 gtgtgtgtgt gtgcgcaatt aaacgttgcg catttctccg cctcttgaat agaggttttg
      361 tttggggttt tttttttttt taagggttta ttctgatgct tttcaagcct cgcttctcct
      421 ttaaggggta aaaaaaaaac accccccccc ccccacctct attaaaaaaa aaaaacaaca
      481 gcaaaaaaac cttcgagacc ttctgttatt cccctctctg agaatgagaa catcagctat
      541 tcaagcgtat tgatgaactc ccccaaaaag tttattaaga gaagcgaaaa caaaaggact
      601 aaaggccgca ggcaactgga cagacagttt ataaactaag gtcccattgt ccccccgagg
      661 cagatgcatg aatacctctg aatgggtccc tttggagtga caagatcaag ccagacaaca
      721 gcatgttaaa aggaaggagc gctgtttctc tttctccagc cgaaagatct gctcccccta
      781 ctcctcctgc attaatgtac atctgcaggg ggaggaataa atagctcctt aggatctgcg
      841 ccgctcaggg tttttattct tccaccccca gcactggctc tttgcaccat atgctccaga
      901 tcagcctcct ttctccgcag cctccagcca agcagtcgcc taaaatgatc cccagccatt
      961 gcaaactttc ttctctccct tttcgccctc ctcctttcct tccctctgcc gctgttaaaa
     1021 tcgcgattta cggcccgctt gcctaggaga tccctcgtgg tcaattaggt ttgcacagag
     1081 caatttaact aataggcttt gatggattta ttttattatt attattattg ttttaccctg
     1141 ggatatcggc cctgttggct agcccgggct cgagatctgc gatctaagta agcttgctgc
     1201 ttcatccccg tggcccgttg ctcgcgtttg ctggcggtgt ccccggaaga aatatatttg
     1261 catgtcttta gttctatgat gacacaaacc ccgcccagcg tcttgtcatt ggcgaattcg
     1321 aacacgcaga tgcagtcggg gcggcgcggt cccaggtcca cttcgcatat taaggtgacg
     1381 cgtgtggcct cgaacaccga gcgaccctgc agcgacccgc ttaacagcgt caacagcgtg
     1441 ccgcagatcc caagcttgcg gccgcttaag ctgcagaagt tggtcgtgag gcactgggca
     1501 ggtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag
     1561 acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc
     1621 ctttctctcc acaggtgtcc actcccaggt tcaatacagc tcttaagcgg ccgcaagcta
     1681 gcttgccgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1741 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1801 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1861 gccctggccc accctcgtga ccaccttcgg ctacggcctg atgtgcttcg cccgctaccc
     1921 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1981 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2041 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2101 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2161 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     2221 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2281 gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg
     2341 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2401 gctgtacaag tgagcctcta gactccctca gcggcgcgcc tggtgttcct cgagatcgga
     2461 gtcctaaatc gactgcctta gcggtaggac agtagatcag tcagcgacgt gacgaaccct
     2521 cgcaggatca agatcccacg tgtctagagt cggggcggcc ggccgcttcg agcagacatg
     2581 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     2641 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     2701 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     2761 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccgat
     2821 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     2881 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     2941 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3001 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3061 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3121 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3181 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3241 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3301 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3361 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3421 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3481 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3541 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3601 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3661 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3721 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3781 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3841 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3901 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3961 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4021 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4081 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4141 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4201 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4261 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4321 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4381 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4441 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4501 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4561 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4621 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4681 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4741 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4801 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4861 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4921 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4981 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5041 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     5101 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5161 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     5221 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     5281 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttgc cattcgccat
     5341 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     5401 ccaagctacc atgataagta agtaatatta aggtacggga ggtacttgga gcggccgc
//
