LOCUS       sequence-249939-        4859 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 4859)
  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 4859)
  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_368_Citrine
FEATURES             Location/Qualifiers
     source          1..4859
                     /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        678..823
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          901..1033
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1016..1035
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1087..1096
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1093..1095,1096..1098,1099..1809)
                     /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(1138..1159)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1399..1418)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1746..1767
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1983..2104
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1994..2013)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2048..2067
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2352..2369)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2523..3111)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2603..2622)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3282..4073,4074..4142))
                     /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     3905..3924
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4143..4247)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4274..4729
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4361..4380)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4571..4592
                     /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 caggaaccct cggctgcctt aaaaatggag taaagggatg ggggagggag ggcttctcta
      241 ttctcattca aaattctgca aggaggagaa aaagttcgga caaagggcca aggaagggaa
      301 gaaagggaga taaattattc aaaactccct gctgttagat agttttgtaa tattatagga
      361 ccgtctgttg gaaaaaaggc gagccgttgg acgttgcaaa atgttgcttg tgatttggaa
      421 ataggcgatg ctcatttgca aatcggggcc attgtgtgcg ggggaaaact gcccccgcgt
      481 caccgcgcga tgcgttgggt ttcattgcgg ggagatggtt tttctccggc tcgcatcgct
      541 cggaaaagaa ctgttggcta gcccgggctc gagatctgcg atctaagtaa gcttgctgct
      601 tcatccccgt ggcccgttgc tcgcgtttgc tggcggtgtc cccggaagaa atatatttgc
      661 atgtctttag ttctatgatg acacaaaccc cgcccagcgt cttgtcattg gcgaattcga
      721 acacgcagat gcagtcgggg cggcgcggtc ccaggtccac ttcgcatatt aaggtgacgc
      781 gtgtggcctc gaacaccgag cgaccctgca gcgacccgct taacagcgtc aacagcgtgc
      841 cgcagatccc aagcttgcgg ccgcttaagc tgcagaagtt ggtcgtgagg cactgggcag
      901 gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga
      961 cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc
     1021 tttctctcca caggtgtcca ctcccaggtt caatacagct cttaagcggc cgcaagctag
     1081 cttgccgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1141 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     1201 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     1261 ccctggccca ccctcgtgac caccttcggc tacggcctga tgtgcttcgc ccgctacccc
     1321 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1381 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1441 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1501 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1561 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1621 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1681 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     1741 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1801 ctgtacaagt gagcctctag actccctcag cggcgcgccg ttgcgttcca aacgtcgtgg
     1861 agctccctgt caagacacga accgctaacg ccaccttagt ccgtgaagca tgatacgcga
     1921 cataccccag gagatctcac gtgtctagag tcggggcggc cggccgcttc gagcagacat
     1981 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2041 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2101 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     2161 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtcgaccga
     2221 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
     2281 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
     2341 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2401 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2461 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2521 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2581 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2641 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2701 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2761 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2821 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2881 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2941 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     3001 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3061 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3121 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3181 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3241 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3301 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3361 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3421 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3481 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3541 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3601 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3661 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3721 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3781 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3841 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3901 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3961 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4021 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4081 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4141 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4201 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4261 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4321 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4381 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4441 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4501 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4561 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4621 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4681 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg ccattcgcca
     4741 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     4801 cccaagctac catgataagt aagtaatatt aaggtacggg aggtacttgg agcggccgc
//
