LOCUS       sequence-258496-        5218 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 5218)
  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 5218)
  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_99a_Citrine
FEATURES             Location/Qualifiers
     source          1..5218
                     /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        990..1135
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1213..1345
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1328..1347
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1399..1408
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1405..1407,1408..1410,1411..2121)
                     /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(1450..1471)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1711..1730)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2058..2079
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2293..2414
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2304..2323)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2358..2377
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2662..2679)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2833..3421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2913..2932)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3592..4383,4384..4452))
                     /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     4215..4234
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4453..4557)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4584..5039
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4671..4690)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4881..4902
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5170..5218
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
       61 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
      121 gctcgagttg ccaggatggc atgcagtggg tttgcggtag gtatgaagga gaaggatgcc
      181 gagcagcctt tggggctgat gataaagagc tgaaggcttt cagacatccg tggtggaggt
      241 agatgagcat gtgggaagct ttgtaccatt caatacctct ttgatgctcc ctctgcatct
      301 ttctttttgc agtgagccct tcccgtgcct cacatcttca ttgcagctgc aggctggggc
      361 agaatactgc atagcaggcc gtctcctctg ccgtttcagt gagtgactgc acaaaggaaa
      421 tggataaaag gagtggaggg tcagagttta attaccctat tgtgtgcaga gatgagtgat
      481 ggacagagac tgtcagtggg tcaccgatta gctgccatga agcagacaga ggaggggaaa
      541 atcgtgatga gcccccagga cagagcaatg ttggggacag ccaagtgata gtgagacatg
      601 gagtaggcag ctcagaatag gagacccttt ttcattcagt tgctcccccc aatccttccg
      661 ctttgttcct gcctccataa ggagccccat gggctccgaa ctctctgctg ggccctgagt
      721 gcttcactgg ccagggtgtg agagctgctt gggtggtagg acctctgggg ggatgtagaa
      781 taaggtgagt cattgccccc gactgcactg agctctcatg tcctggtggg gcggaggcaa
      841 ggaaagaggg aaggaaggca ggatgttggc tagcccgggc tcgagatctg cgatctaagt
      901 aagcttgctg cttcatcccc gtggcccgtt gctcgcgttt gctggcggtg tccccggaag
      961 aaatatattt gcatgtcttt agttctatga tgacacaaac cccgcccagc gtcttgtcat
     1021 tggcgaattc gaacacgcag atgcagtcgg ggcggcgcgg tcccaggtcc acttcgcata
     1081 ttaaggtgac gcgtgtggcc tcgaacaccg agcgaccctg cagcgacccg cttaacagcg
     1141 tcaacagcgt gccgcagatc ccaagcttgc ggccgcttaa gctgcagaag ttggtcgtga
     1201 ggcactgggc aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg
     1261 ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac
     1321 atccactttg cctttctctc cacaggtgtc cactcccagg ttcaatacag ctcttaagcg
     1381 gccgcaagct agcttgccgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     1441 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1501 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     1561 aagctgcccg tgccctggcc caccctcgtg accaccttcg gctacggcct gatgtgcttc
     1621 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1681 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1741 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1801 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     1861 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     1921 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1981 cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgag caaagacccc
     2041 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     2101 ggcatggacg agctgtacaa gtgagcctct agactccctc agcggcgcgc ctggtgttcc
     2161 tcgagatcgg agtcctaaat cgactgcctt agcggtagga cagtagatca gtcagcgacg
     2221 tgacgaaccc tcgcaggatc aagatcccac gtgtctagag tcggggcggc cggccgcttc
     2281 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     2341 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     2401 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     2461 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc
     2521 cgtcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
     2581 atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg
     2641 ccggcagcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2701 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2761 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2821 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2881 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2941 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3001 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3061 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3121 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3181 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3241 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3301 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3361 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3421 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3481 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3541 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3601 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3661 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3721 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3781 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3841 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3901 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3961 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4021 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4081 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4141 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4201 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4261 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4321 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4381 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4441 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4501 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4561 atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4621 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4681 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4741 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4801 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4861 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4921 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4981 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg
     5041 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     5101 attacgccag cccaagctac catgataagt aagtaatatt aaggtacggg aggtacttgg
     5161 agcggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtg
//
