LOCUS       sequence-258501-        6393 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 6393)
  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 6393)
  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_85_Citrine
FEATURES             Location/Qualifiers
     source          1..6393
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2060..2205
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2283..2415
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2398..2417
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2469..2478
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2475..2477,2478..2480,2481..3191)
                     /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(2520..2541)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2781..2800)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3128..3149
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3363..3484
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3374..3393)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3428..3447
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3732..3749)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3903..4491)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3983..4002)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4662..5453,5454..5522))
                     /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     5285..5304
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5523..5627)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5654..6109
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5741..5760)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5951..5972
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6240..6288
                     /note="synthetic polyadenylation signal"
     misc_feature    6302..6393
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6342..6361
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 atcgataggt accgagctcg agttgccagg gggttttcgc tgtgcacttc tcagcactgt
       61 ttgcatcagc tcatttccac aggtagcaac aagcaaagtg aagatgagca aagccgtggt
      121 ggctgcactg gttttgtcat cttgtttagt cagggtgaca aaagcaggtt caatctcgag
      181 gaacgcggag aacagccctc ctcctgcttc cctcgccccc acagtcccaa tgaatagagt
      241 tggcaagggt gggcacagcc ccagccactg gcgctgatgg ctctcgacgt gaataagcag
      301 cataaaggca cagagcggcc ccgtcgcggc tccgtcccga cccatcctcc ccagctgcag
      361 cagcatccgc tgcgagacca gaccggccat tcctaccccg tctttctgtg tagttcatct
      421 ttttctcccc ctctcatctc cccccacctc cccctctctc tctttctctc tccctctctt
      481 tctctctctc tctttttttt tttttaattt aatccccttt tttttccgaa catatcatta
      541 caggagcaac aaaaaagatg ggacaataca gccattgatg agctgctctc gtcctcccgt
      601 cccagtcccc cccactccct ccctcccctc ctctggtgcg tgccaacact ctttccaccc
      661 gcccagggct ggagccagag ccaaacaacc tctgctaata aacaagagga accagataaa
      721 ccaggaacac aatagaggga tttgttgcac tttagtgatg gtgggaacaa tgtcaactcc
      781 gtggcgcagc tgcatggacc ccctcccatc ctctcccctc actgccttgt gcccccctcg
      841 cactcgcagc ccaacactgt gatttatgaa aggccagagg tcactttgca ttaaaggggt
      901 cctttcttgc atcatttatt aacccctcta cgacaagggg tgcttattgt ggcagtggtg
      961 gcacagggat ggagaggggg cggaggagtc catcccagag gcactgcaca tctcttgtag
     1021 gatgctgaga tgctgagcta gtgcttaaat ccattttcac agcaacctga ggcacatggg
     1081 aaggaaggaa agctttagtg ggatgcattt tggctaccag ctgtgtgcca ataaaagcaa
     1141 cttgtgcaat ccagtgtaga tctgttccca tttcacacgg gtcatcttcc acccaacctg
     1201 tgctcgatgc cgtgctggtg agccgaactc atgtttagtc ctgcagctac gcatgcaccc
     1261 aggactggcc tgaaggattg ccctgcagac cacatcagcc atatggtggt cctgcaggac
     1321 gaacctgggt atgtcactgc agccttggga caccttgtgc cagcaggtgt gaacctgtag
     1381 cgcatccagg cagagaggtg gcaggttggc cacctcaaaa tccccttctc aggccaggat
     1441 agaggtggtc aaaatctctc cttgccttgt ggaagcaggt ttccatcaca ggcaatgctg
     1501 cactgtcacc acgcaatgct ttcaagcctc tctttgtaac tgcggagcaa aacaatgaaa
     1561 catggacgtg ctaatttgca ggcttggtct tttgttgcag tgaccacggt cactttggga
     1621 gaactgatgt ggtggcaaag cactgaactc tacagtcccc tgagctaccc acatcctcag
     1681 atgtgcttgt gggctccttg gagaccctca ccctggccaa gcttgctagg aaagacaaaa
     1741 cccatccctt ctgatgacct tatgttttgg tgcaggggat acaatggtgc tggtgggctg
     1801 aggcatcttc cctctcatga aatagatggg gaacaaggga caaaagaact ggaaaccagt
     1861 ccccaaatta gggcctggtg tgagcccttt gtgtatgggc tcacagggca gaggggatgt
     1921 aaaggttggg gtctgttggc tagcccgggc tcgagatctg cgatctaagt aagcttgctg
     1981 cttcatcccc gtggcccgtt gctcgcgttt gctggcggtg tccccggaag aaatatattt
     2041 gcatgtcttt agttctatga tgacacaaac cccgcccagc gtcttgtcat tggcgaattc
     2101 gaacacgcag atgcagtcgg ggcggcgcgg tcccaggtcc acttcgcata ttaaggtgac
     2161 gcgtgtggcc tcgaacaccg agcgaccctg cagcgacccg cttaacagcg tcaacagcgt
     2221 gccgcagatc ccaagcttgc ggccgcttaa gctgcagaag ttggtcgtga ggcactgggc
     2281 aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga
     2341 gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg
     2401 cctttctctc cacaggtgtc cactcccagg ttcaatacag ctcttaagcg gccgcaagct
     2461 agcttgccgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     2521 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     2581 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     2641 tgccctggcc caccctcgtg accaccttcg gctacggcct gatgtgcttc gcccgctacc
     2701 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2761 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2821 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2881 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     2941 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     3001 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     3061 tgcccgacaa ccactacctg agctaccagt ccgccctgag caaagacccc aacgagaagc
     3121 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     3181 agctgtacaa gtgagcctct agactccctc agcggcgcgc ctggtgttcc tcgagatcgg
     3241 agtcctaaat cgactgcctt agcggtagga cagtagatca gtcagcgacg tgacgaaccc
     3301 tcgcaggatc aagatcccac gtgtctagag tcggggcggc cggccgcttc gagcagacat
     3361 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     3421 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     3481 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     3541 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtcgaccga
     3601 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
     3661 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
     3721 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3781 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3841 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3901 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3961 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4021 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4081 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4141 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4201 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4261 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4321 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4381 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4441 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4501 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4561 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4621 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4681 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4741 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4801 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4861 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4921 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4981 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5041 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5101 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5161 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5221 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5281 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5341 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5401 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5461 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5521 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5581 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5641 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5701 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5761 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5821 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     5881 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     5941 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     6001 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     6061 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg ccattcgcca
     6121 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     6181 cccaagctac catgataagt aagtaatatt aaggtacggg aggtacttgg agcggccgca
     6241 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac
     6301 taacatacgc tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc
     6361 ccagtgcaag tgcaggtgcc agaacatttc tct
//
