LOCUS       sequence-258511-        7304 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 7304)
  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 7304)
  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_195_Citrine
FEATURES             Location/Qualifiers
     source          1..7304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2779..2924
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          3002..3134
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3117..3136
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      3188..3197
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3194..3196,3197..3199,3200..3910)
                     /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(3239..3260)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3500..3519)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3847..3868
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4084..4205
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4095..4114)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4149..4168
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4453..4470)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4624..5212)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4704..4723)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5383..6174,6175..6243))
                     /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     6006..6025
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6244..6348)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6375..6830
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6462..6481)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6672..6693
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6961..7009
                     /note="synthetic polyadenylation signal"
     misc_feature    7023..7114
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7063..7082
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 cccccctttc ccagcctcac tttcccctct gcagcggaag atttgctatt tcatggctgg
       61 ctaaagccaa acttccctca agggggttat gtgactcgtg accccctcta acctgtactt
      121 agtgctctcc aaagggtgtg tactcctgga gaggcgacct gaatgcaggc tgagccacga
      181 gggcaaaacc aacctattaa cagaaagctg aaacagcagc ggggaggaaa aaacaccgag
      241 caaacatctc aagtataaaa ctaaaacaga agtgattctg cattggggaa atacctgttt
      301 catcagcttg ggcagtggta tccaaagagc agtaggaacc tgaagcgtag caggcaagat
      361 gccctatttc caggcatggt tcagaggtgg ccatgggatt ttactcacat cagtaatccc
      421 tttgccttcc gtggacctac tgatgggcac aggttgctgc aggacctgca aaagtgtttg
      481 aggtccttat gtacttttat tatttccttt ggggcttcca tggggtccct ttgcccgaaa
      541 gcactgcaga tggggcaggg ggagagggga tgaaaaaaga aacagatgag gaaccgatgt
      601 gatgaaagag cagcagattt tgtgtcgttc ctgtccccac agtggctccc tgaggcattt
      661 caaaggggag aaaaaaggaa ttgggaaaac acaaacgcat aatcaaacca aggagccagt
      721 taaccaggcc aatacccgga atccccaagt cccatttagc tgctcgcagg attatcactt
      781 gttgctgatt caaaaccagc aggtgttgtc agctatttat gatcactgtt atttatacag
      841 tgccactggt gtgcctggcg cacggcagac aggcagatga gagccaacct ctcccctgag
      901 aagataccaa actaaattag actcaacaca gaggcagacg ttaatacgca gcatatgggg
      961 acgaggggaa aaaggaggtt caaatccaca aagatcgtga aatctccagg cattgaatcc
     1021 cacacccatc agctggttgg ttcccacctt gtaggaaaag caaagaagca gttccatatt
     1081 tacacacggg tagggaccaa ggcaaggtgc agaaccttac tttgcaggca gagtggatgg
     1141 atgctgtttg ctttagggga tggggtgacg ttagagctgg ggataagcaa agagaagcca
     1201 cttgctcagg ttcgcatgga gtatcagcag caaagccagg gctggagcct cctgtaagaa
     1261 ttactaagca gagctgtatc cacagccttc ttcttcataa cctgagggtt ttatgcagga
     1321 aaatacccat ggcatctgct ggggttatag attgaatacg gacctggaga tttggctttt
     1381 attattatta tttttgttat ctttagaggc tcgtggagca aacaacgaga tgttacttgg
     1441 aaataaacca cgtttccagc ttaggcctgc aaagacttac atttgcgggt gactttatcc
     1501 aaatgagtca tcccatccca tgtctgtgag aggattcccc gtgtaagcaa aattgactaa
     1561 cacgacgttt gtagggattg ggccaggcac cggtaagagg cattccccag cttccagcaa
     1621 ggaggctgca tggggaaagg aaatcagctc tgcagaaccc cagcagcctc ctcctgcctt
     1681 gctcttaaaa gacaaaagtc acaaacctgg tgactggttg ctgcttagct ggcaggggtg
     1741 taaaatgcaa agtaaccccc cgtagttatt tgaagaaggc ttggtcctgc tgtaaagctc
     1801 tataaactgg gctcctttcc ccatcccctc ctccccggca tgttttaaat gattttggat
     1861 tttaatgcag ggcccggagc tcattccacc gtattcatgg catcaaacaa gaggctgctg
     1921 cccccagctg ggttccttga acttagaagc aaagctctac tcaatattac cttaattaat
     1981 taactaaaga gacaatggct tttgaggtct cacctctgtt tccagggagg ggggtctttt
     2041 gacagagccc tgccataaac taattcagag ttccagcaag gagtcatagg gggaagagtt
     2101 aaatgacact atttacttgt ttaccaagac aatttatgtc atatttgttt ttccccacct
     2161 tctgtctgct gcactgagaa ctctgtcggg ggtgcagggg gaaggaggag gaggagagag
     2221 tattatttgc attaaccact gaaatcaaag catattccca tctgaaaaat accacttcat
     2281 agctcttggt atcttcttct tctttttttt tttaatttta atttacaaaa agtttttgaa
     2341 gctttccagg ttcaatataa caaatctgag ctttgatgct ttaaaactgc cataaagagc
     2401 tacaggcaac atcacagtta accaaaaaat atatatatat atttcaaggc tcctgggagt
     2461 acttgcatct cctctgaatg ttgtggggtt tttttttgtt catttgcaag caagcactga
     2521 gtcgttggtt tattccactc agaaacaggc atctcgtgtt ttgtatcagc aagaaaaagc
     2581 ttttacaagg tcacaatgaa aagtgaccgt ctggaaaatc tatctaggaa cacgtaggga
     2641 acacatggca gctgttggct agcccgggct cgagatctgc gatctaagta agcttgctgc
     2701 ttcatccccg tggcccgttg ctcgcgtttg ctggcggtgt ccccggaaga aatatatttg
     2761 catgtcttta gttctatgat gacacaaacc ccgcccagcg tcttgtcatt ggcgaattcg
     2821 aacacgcaga tgcagtcggg gcggcgcggt cccaggtcca cttcgcatat taaggtgacg
     2881 cgtgtggcct cgaacaccga gcgaccctgc agcgacccgc ttaacagcgt caacagcgtg
     2941 ccgcagatcc caagcttgcg gccgcttaag ctgcagaagt tggtcgtgag gcactgggca
     3001 ggtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag
     3061 acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc
     3121 ctttctctcc acaggtgtcc actcccaggt tcaatacagc tcttaagcgg ccgcaagcta
     3181 gcttgccgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     3241 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     3301 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     3361 gccctggccc accctcgtga ccaccttcgg ctacggcctg atgtgcttcg cccgctaccc
     3421 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     3481 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     3541 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     3601 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     3661 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     3721 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     3781 gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg
     3841 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     3901 gctgtacaag tgagcctcta gactccctca gcggcgcgcc gttgcgttcc aaacgtcgtg
     3961 gagctccctg tcaagacacg aaccgctaac gccaccttag tccgtgaagc atgatacgcg
     4021 acatacccca ggagatctca cgtgtctaga gtcggggcgg ccggccgctt cgagcagaca
     4081 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     4141 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     4201 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     4261 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat ccgtcgaccg
     4321 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc
     4381 gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
     4441 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4501 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4561 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4621 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4681 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4741 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4801 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4861 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4921 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4981 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5041 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5101 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5161 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5221 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5281 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5341 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5401 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5461 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5521 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5581 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5641 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5701 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5761 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5821 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5881 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5941 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6001 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6061 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6121 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6181 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6241 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6301 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6361 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     6421 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     6481 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     6541 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     6601 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     6661 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     6721 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     6781 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt gccattcgcc
     6841 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     6901 gcccaagcta ccatgataag taagtaatat taaggtacgg gaggtacttg gagcggccgc
     6961 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta
     7021 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
     7081 cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat aggtaccgag ctcgagttgc
     7141 cagggggtgt atggggctgg taagtgccgt gatggatcgg tccggggtgt ccctggtgat
     7201 ggaggggcca tatgcctgtt tgcatgctcc taacaagagg cagctctgag tgttgggttg
     7261 ggaggtcaag cgatgataat aataataaaa ttaccccccc cccc
//
