LOCUS       sequence-258514-        5313 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 5313)
  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 5313)
  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_Sox9_enh_349_Citrine
FEATURES             Location/Qualifiers
     source          1..5313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    14..105
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     54..73
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        1083..1228
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1306..1438
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1421..1440
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1492..1501
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1498..1500,1501..1503,1504..2214)
                     /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(1543..1564)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1804..1823)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2151..2172
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2388..2509
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2399..2418)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2453..2472
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2757..2774)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2928..3516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3008..3027)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3687..4478,4479..4547))
                     /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     4310..4329
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4548..4652)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4679..5134
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4766..4785)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4976..4997
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5265..5313
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
       61 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
      121 gctcgagttg ccaggaaaga gcagggaagg atggcaagcc ctgggagcga tgctgccaac
      181 catcagcatc tcaacttcta gggcatccca gcttttcagc ttgcacggag cagctcagcc
      241 ccacgccctc cctgctcctc ctccagcaca gcattaagca cggtgtcatt ccactgcggg
      301 agcctgctct aattaaagtg tcagcctttc cgccggcagg gatgataatt gaggtagagc
      361 gttacagtgt tttcaaaccc tgtgctgcag gcggagagct cagcagatct gggagtgcca
      421 ccaggcactg gtgacaccgg cagcgtgggg acggcactgg gatggcaaag cagaagagct
      481 ctggggagaa ccaaacccag cagcatcctg taggcagctc ggagaccatg agcctgacct
      541 cactgcgctc tgcccccaac ccatgggctg gacacaaagc agcagcaggc aatggaccgg
      601 gttcagggcc cccactggca cagctcccca aatattaagc tttaatgttg tgacctttaa
      661 gatgcaaaag aatgtctgag ctcctgcaac tgcccaaagg tggctttgtg ctcttgccga
      721 agcaatgaaa tgttggatct gtgtctctgc agcaacctgg ggtgtgcact cattaactca
      781 gtcctgcggg tgatgctttg ttcattcctc tacaaaactc gctttttgca atgacagtga
      841 aaggaatccc ccaccaacac agcctgaggc agtgagctgc tcacacagcc cagctcctca
      901 ctccaggctc cctggcactt catttgcagg atgcaatcca attaccgctc accccctgtt
      961 ggctagcccg ggctcgagat ctgcgatcta agtaagcttg ctgcttcatc cccgtggccc
     1021 gttgctcgcg tttgctggcg gtgtccccgg aagaaatata tttgcatgtc tttagttcta
     1081 tgatgacaca aaccccgccc agcgtcttgt cattggcgaa ttcgaacacg cagatgcagt
     1141 cggggcggcg cggtcccagg tccacttcgc atattaaggt gacgcgtgtg gcctcgaaca
     1201 ccgagcgacc ctgcagcgac ccgcttaaca gcgtcaacag cgtgccgcag atcccaagct
     1261 tgcggccgct taagctgcag aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt
     1321 acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg
     1381 cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt
     1441 gtccactccc aggttcaata cagctcttaa gcggccgcaa gctagcttgc cgccaccatg
     1501 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     1561 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     1621 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1681 gtgaccacct tcggctacgg cctgatgtgc ttcgcccgct accccgacca catgaagcag
     1741 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     1801 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1861 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1921 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     1981 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     2041 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     2101 ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     2161 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtgagcc
     2221 tctagactcc ctcagcggcg cgccgttgcg ttccaaacgt cgtggagctc cctgtcaaga
     2281 cacgaaccgc taacgccacc ttagtccgtg aagcatgata cgcgacatac cccaggagat
     2341 ctcacgtgtc tagagtcggg gcggccggcc gcttcgagca gacatgataa gatacattga
     2401 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     2461 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     2521 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     2581 aaacctctac aaatgtggta aaatcgataa ggatccgtcg accgatgccc ttgagagcct
     2641 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     2701 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg
     2761 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2821 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2881 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2941 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3001 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3061 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3121 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3181 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3241 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3301 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3361 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3421 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3481 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3541 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3601 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3661 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3721 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3781 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3841 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3901 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3961 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4021 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4081 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4141 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4201 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4261 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4321 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4381 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4441 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4501 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4561 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4621 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     4681 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4741 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4801 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4861 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4921 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4981 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5041 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5101 gcgaatttta acaaaatatt aacgcttaca atttgccatt cgccattcag gctgcgcaac
     5161 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagcccaa gctaccatga
     5221 taagtaagta atattaaggt acgggaggta cttggagcgg ccgcaataaa atatctttat
     5281 tttcattaca tctgtgtgtt ggttttttgt gtg
//
