LOCUS       sequence-259022-        5837 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 5837)
  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 5837)
  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_143_Citrine
FEATURES             Location/Qualifiers
     source          1..5837
                     /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        1608..1753
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1831..1963
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1946..1965
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2017..2026
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2023..2025,2026..2028,2029..2739)
                     /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(2068..2089)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2329..2348)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2676..2697
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2912..3033
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2923..2942)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2977..2996
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3281..3298)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3452..4040)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3532..3551)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4211..5002,5003..5071))
                     /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     4834..4853
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5072..5176)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5203..5658
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5290..5309)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5500..5521
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5789..5837
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
       61 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
      121 gctcgagttg ccagggggtt gggaaaggtt tatgaggtta aatttgttta tctttcctga
      181 aagagtctta tattgaaagg tctttaactt gatgtatcca gccagggagg ccagggctgg
      241 aacagcagag ggactacagg acaagattaa gggccattgg cagagctgta taggagatgg
      301 ggatgaaaat aaaagagtgt gttgcaaatt gggacgcagg agcagcgtga tagagctgtg
      361 caggggcccc agggtagcaa tagcaaggca gctatcagcc aggattgaga tgtgttggca
      421 aagcatgcag taacaggacg gggacaggag ggatggcagg actgcagagg ttttcacagg
      481 caaaacctgg gtctgtgcac ctgggtagga tgctgggcag cagagacagc acgtctgtgt
      541 gtatgtgatg tactgagcgt gctataccaa gtggttttca ctccagatct ggaggaaggt
      601 gaatttgact ttagctatct gagacagtgc ttcttgctcg gtcagccggg ggaaaattcc
      661 gagcaaaaag tgctggaaga ggctgaggac aaagatcaga accaaggaac aggggcagga
      721 cagagcccag ccagatgaca aacacctctg ggcagttggg agtctcgctc ccgtggatct
      781 gagttgtttt tgtgcaggtg tttggatagt gcagtgtctt ttctatgatg gtgtttgtac
      841 ctccaggcgc cctcagaggg tggtcgtaac acatgcaggg agcactcagc tattacattg
      901 tgcttggagt gaggagtcca gctggtgtca tgcgctcctg gaggctcacc cccttattac
      961 agtggctgcg atgttaggga tgctttgctt gctgctatgg gagaacaggc atggctggag
     1021 catgggctga gagcgaggag gggctgcgtg gtgcttgggg ctgcgatgta ttgctgcccc
     1081 gctaccagct ggtttccctt tgcttcccat ttttgggagc actatcgatg cctctccgac
     1141 tgcatcgttt caattcaagt cgggcagcca gcaagcccct gcagtccctc cggcgcagcc
     1201 ctccgctgga ccccgtgcct gccacaaaca cgcagctatg aatattgaag tgaaaaaggg
     1261 cccttttatg gacttcacag atgtgctggg gagttggggc aggcctcatt acctcggggt
     1321 tctttaaaag gccaaacaaa aaccagaaga aagagtgtta agggtagagg ggggcaggga
     1381 ggcgagaact agaaaaaagt tgaaagaaaa gaaaagacga ctggagttga tggatattcc
     1441 atctgtttat gttcagacca gtgctgggaa ttaacaaagg ctgttggcta gcccgggctc
     1501 gagatctgcg atctaagtaa gcttgctgct tcatccccgt ggcccgttgc tcgcgtttgc
     1561 tggcggtgtc cccggaagaa atatatttgc atgtctttag ttctatgatg acacaaaccc
     1621 cgcccagcgt cttgtcattg gcgaattcga acacgcagat gcagtcgggg cggcgcggtc
     1681 ccaggtccac ttcgcatatt aaggtgacgc gtgtggcctc gaacaccgag cgaccctgca
     1741 gcgacccgct taacagcgtc aacagcgtgc cgcagatccc aagcttgcgg ccgcttaagc
     1801 tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa
     1861 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
     1921 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccaggtt
     1981 caatacagct cttaagcggc cgcaagctag cttgccgcca ccatggtgag caagggcgag
     2041 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     2101 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     2161 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc
     2221 tacggcctga tgtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     2281 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     2341 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     2401 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     2461 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     2521 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     2581 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     2641 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     2701 gccgccggga tcactctcgg catggacgag ctgtacaagt gagcctctag actccctcag
     2761 cggcgcgcct actcgtccgg cgtcacaaat tagcggaccg accactgcac ctctagcgta
     2821 ccgtcaaagg gcaacgaggg tcattccgtg ccgacataag agatctcacg tgtctagagt
     2881 cggggcggcc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     2941 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     3001 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     3061 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     3121 ggtaaaatcg ataaggatcc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc
     3181 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     3241 gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc
     3301 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3361 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3421 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3481 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3541 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3601 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3661 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3721 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3781 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3841 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3901 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3961 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4021 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4081 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4141 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4201 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4261 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4321 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4381 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4441 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4501 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4561 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4621 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4681 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4741 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4801 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4861 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4921 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4981 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5041 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5101 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5161 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
     5221 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     5281 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5341 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5401 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5461 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5521 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     5581 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5641 tattaacgct tacaatttgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     5701 cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc atgataagta agtaatatta
     5761 aggtacggga ggtacttgga gcggccgcaa taaaatatct ttattttcat tacatctgtg
     5821 tgttggtttt ttgtgtg
//
