LOCUS       sequence-259025-        5360 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 5360)
  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 5360)
  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_218_Citrine
FEATURES             Location/Qualifiers
     source          1..5360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        888..1033
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1111..1243
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1226..1245
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1297..1306
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1303..1305,1306..1308,1309..2019)
                     /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(1348..1369)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1609..1628)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1956..1977
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2191..2312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2202..2221)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2256..2275
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2560..2577)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2731..3319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2811..2830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3490..4281,4282..4350))
                     /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     4113..4132
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4351..4455)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4482..4937
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4569..4588)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4779..4800
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5068..5116
                     /note="synthetic polyadenylation signal"
     misc_feature    5130..5221
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5170..5189
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 gtgcaggctt gaaaactcaa aggagggcag ggtaagtgag ctccgtgctg ctcgtccaaa
       61 gctcgctcct ccatccctgc cccagtgctc agagttagac tgattttcca ggctttttaa
      121 ctcctcctcc gcaggggccc aaccagccct gctcccaaat tccccccccc cagacaaatc
      181 tgggaaggat ctcatgccgg cagagtcggg aggtgggatc ccatcaaggc aaagcggctt
      241 tgcaaggctc cggggccctt ccggggggag gagaagcttt agaaagcaaa gggcagtgca
      301 tcccctaagc cccagtccag caggaaggtg agggaggagg ggacagagac gaggaggaag
      361 gggacccggc gctctataca acaactcttg tgtccttcca gagaatggat gctcctcaca
      421 atagctcacc aggccactgg cgggggataa agggctcatt ttaccacttg ccaggctcca
      481 aatatttcca atttgccact ttaattgatg tcaataaggt tatgctaaag tcggggattt
      541 gcagccatcc cagcgctgcc gtggcccccc gaggtgacgg gttgtcccct cacctccccc
      601 agcacgaatt cagcagcgcc ggtccagcaa actactcaca cccttaaccg ggcacccagg
      661 ggtcactttg ctgattgcag gaggcaattg cattaaggca gtatattggg atggaggcct
      721 gtaggcagtg gcttgcccca cgctcaccca caggaacaaa ctgttggcta gcccgggctc
      781 gagatctgcg atctaagtaa gcttgctgct tcatccccgt ggcccgttgc tcgcgtttgc
      841 tggcggtgtc cccggaagaa atatatttgc atgtctttag ttctatgatg acacaaaccc
      901 cgcccagcgt cttgtcattg gcgaattcga acacgcagat gcagtcgggg cggcgcggtc
      961 ccaggtccac ttcgcatatt aaggtgacgc gtgtggcctc gaacaccgag cgaccctgca
     1021 gcgacccgct taacagcgtc aacagcgtgc cgcagatccc aagcttgcgg ccgcttaagc
     1081 tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa
     1141 ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca
     1201 cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccaggtt
     1261 caatacagct cttaagcggc cgcaagctag cttgccgcca ccatggtgag caagggcgag
     1321 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     1381 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     1441 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc
     1501 tacggcctga tgtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     1561 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     1621 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1681 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1741 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     1801 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1861 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     1921 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1981 gccgccggga tcactctcgg catggacgag ctgtacaagt gagcctctag actccctcag
     2041 cggcgcgcct ggtgttcctc gagatcggag tcctaaatcg actgccttag cggtaggaca
     2101 gtagatcagt cagcgacgtg acgaaccctc gcaggatcaa gatcccacgt gtctagagtc
     2161 ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     2221 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2281 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     2341 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     2401 gtaaaatcga taaggatccg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     2461 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     2521 caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg
     2581 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2641 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2701 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2761 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2821 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2881 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2941 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3001 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3061 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3121 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3181 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3241 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3301 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3361 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3421 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3481 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3541 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3601 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3661 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3721 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3781 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3841 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3901 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3961 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4021 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4081 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4141 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4201 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4261 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4321 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4381 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4441 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc
     4501 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4561 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4621 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4681 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4741 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4801 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4861 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4921 attaacgctt acaatttgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     4981 ggtgcgggcc tcttcgctat tacgccagcc caagctacca tgataagtaa gtaatattaa
     5041 ggtacgggag gtacttggag cggccgcaat aaaatatctt tattttcatt acatctgtgt
     5101 gttggttttt tgtgtgaatc gatagtacta acatacgctc tccatcaaaa caaaacgaaa
     5161 caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc
     5221 tatcgatagg taccgagctc gagttgccag ggcttcattg aggtgatgtg cctggagggc
     5281 tgggggggat ttagggtccg atggatgaaa cagatccatt gctttttaac tagtgtggac
     5341 cagcagcaag cctaagctcg
//
