LOCUS       sequence-258626-        6928 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 6928)
  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 6928)
  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_99_Citrine
FEATURES             Location/Qualifiers
     source          1..6928
                     /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        2700..2845
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2923..3055
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3038..3057
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      3109..3118
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3115..3117,3118..3120,3121..3831)
                     /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(3160..3181)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3421..3440)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3768..3789
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4003..4124
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4014..4033)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4068..4087
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4372..4389)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4543..5131)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4623..4642)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5302..6093,6094..6162))
                     /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     5925..5944
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6163..6267)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6294..6749
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6381..6400)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6591..6612
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6880..6928
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
       61 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
      121 gctcgagttg ccagggtggc tccctaactg cttgtttacc cacctgcaaa aaatatcact
      181 gccgcactgg acatcttagg acctccaggg atgaggtgta gcttctgtcc actggcaagc
      241 tggaaatatc cctccacact ggatcacctt ggcattgttt cacaaatttc ttctcctact
      301 caaagatata agtctcaggt gcatctgttc ttacctaccg ctaatatact attcagtcct
      361 taccacagaa tcataggggt tgaaaggggc ctgtggagat caagtccaac tccttgctaa
      421 agcggattcc ctacagtagg ttacacagga aagcacccac atgggtctcg aatatctcca
      481 gcgaagaaga ctgtgcaacc tctctgggct gccttatccc ctagaaggtc cagctgttag
      541 ttagatcaag gcattagccg tggaaatgca atatttgcaa cttgaaaaac cagcgatgta
      601 aagcctaacg aggtttctga gaccctgcag gtggaaagtt tgggatcttg gcctttgcac
      661 accaaggaca cgatcacact ttattgcaca gtctagaggg actgtccatc ttgcaaggtg
      721 ctaccaaagc tcacggtgct ttggttaagc cctgagggtg tagagcccgg tggtgaaaga
      781 ggaacggggc atataggggg ccgcatttca tgagcagtgg cagagtgtag gatgggaaac
      841 aggcagagat ttgccctcat tatgcctggg tctaacatgt ccaattaccc catcctctcc
      901 tcacctcctg gatgtaaaag actcttgttg agagtggcca taattcacag tgaacaccct
      961 gcctccccct tactcccttc acacccgctg ctctccattt tccttccaga aaaaaatatg
     1021 tctttgatga aatgaaatgg ctcagtcaaa gaacccctca aatggttggg gacccctgca
     1081 cttggccacc cttgcccact tcccagctct atttctggcc gagtttggag tttaagtagg
     1141 ctgaccttgt gccccggctg ccctgggggt cagtgatgaa agcggatgga ggcttcacag
     1201 tcaggtctct tgtgtgtgac tcttttgttc ccggacaggg gtgtgcagtg cctgggacag
     1261 tgcttaattg ttcaggggca ctgctttacc cgtcatgccc ttccccgggc ggccccagcc
     1321 cttcttgctc atgtgtggga gggaggcggc ttctccactt ggggtaggga ggaggaagca
     1381 cagtctgggt tgctcggcat aatttctggc tgtttattaa tccatgtaat aaattttaga
     1441 gtgtcaggaa gcaaaaataa aaataagaag aaaaaaaaaa aaaggaattc ctggcatcat
     1501 tcacaattcc tgtttgtcac tgggtgcacc ctcccttgtc ctgttgcttg gtctatctgt
     1561 cgtgcttcat ggatgcaccc cggcaccagg ggttgaggga aacgaggttg ggcttagcct
     1621 gagaggaggt ggtgggcttg tgacagggag tgcagaagcc ttgggttgga gcacactgat
     1681 aagacaaggt ggtggtgtgg aaaagtgagt ccaagactgg aggtgttgaa gctcctggac
     1741 aactttgcta ggcccaggct gagcacccac acctggcaat gccctgatgg catgcagtgg
     1801 gtttgcggta ggtatgaagg agaaggatgc cgagcagcct ttggggctga tgataaagag
     1861 ctgaaggctt tcagacatcc gtggtggagg tagatgagca tgtgggaagc tttgtaccat
     1921 tcaatacctc tttggtgctc cctctgcatc tttctttttg cagtgagccc ttcccgtgcc
     1981 tcacatcttc attgcagctg caggctgggg cagaatactg catagcaggc cgtctcctct
     2041 gccgtttcag tgagtgactg cacaaaggaa atggataaaa ggagtggagg gtcagagttt
     2101 aattacccta ttgtgtgcag agatgagtga tggacagaga ctgtcagtgg gtcaccgatt
     2161 agctgccatg aagcagacag aggaggggaa aatcgtgatg agcccccagg acagagcaat
     2221 gttggggaca gccaagtgat agtgagacat ggagtaggca gctcagaata ggagaccctt
     2281 tttcattcag ttgctccccc caatccttcc gctttgttcc tgcctccata aggagcccca
     2341 tgggctccga actctctgct gggccctgag tgcttcactg gccagggcgt gagagctgct
     2401 tgggtggtag gacctctggg gggatgtaga ataaggtgag tcattgcccc cgactgcact
     2461 gagctctcat gtcctggtgg ggcggaggca aggaaagagg gaaggaaggc aggagggaag
     2521 gtaaggaaga aggctgaagg aggtggctgc tggtgggtga ggtgtggtac tgctgttggc
     2581 tagcccgggc tcgagatctg cgatctaagt aagcttgctg cttcatcccc gtggcccgtt
     2641 gctcgcgttt gctggcggtg tccccggaag aaatatattt gcatgtcttt agttctatga
     2701 tgacacaaac cccgcccagc gtcttgtcat tggcgaattc gaacacgcag atgcagtcgg
     2761 ggcggcgcgg tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc tcgaacaccg
     2821 agcgaccctg cagcgacccg cttaacagcg tcaacagcgt gccgcagatc ccaagcttgc
     2881 ggccgcttaa gctgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca
     2941 agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt
     3001 ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc
     3061 cactcccagg ttcaatacag ctcttaagcg gccgcaagct agcttgccgc caccatggtg
     3121 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     3181 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     3241 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     3301 accaccttcg gctacggcct gatgtgcttc gcccgctacc ccgaccacat gaagcagcac
     3361 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     3421 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     3481 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     3541 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     3601 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     3661 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     3721 agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     3781 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtgagcctct
     3841 agactccctc agcggcgcgc ctggtgttcc tcgagatcgg agtcctaaat cgactgcctt
     3901 agcggtagga cagtagatca gtcagcgacg tgacgaaccc tcgcaggatc aagatcccac
     3961 gtgtctagag tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt
     4021 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     4081 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     4141 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     4201 tctacaaatg tggtaaaatc gataaggatc cgtcgaccga tgcccttgag agccttcaac
     4261 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc
     4321 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac
     4381 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4441 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4501 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4561 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4621 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4681 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4741 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4801 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4861 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4921 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4981 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5041 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5101 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5161 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5221 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5281 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5341 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5401 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5461 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5521 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5581 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5641 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5701 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5761 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5821 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5881 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5941 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6001 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6061 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6121 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6181 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6241 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc
     6301 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     6361 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     6421 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     6481 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     6541 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     6601 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     6661 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     6721 ttttaacaaa atattaacgc ttacaatttg ccattcgcca ttcaggctgc gcaactgttg
     6781 ggaagggcga tcggtgcggg cctcttcgct attacgccag cccaagctac catgataagt
     6841 aagtaatatt aaggtacggg aggtacttgg agcggccgca ataaaatatc tttattttca
     6901 ttacatctgt gtgttggttt tttgtgtg
//
