LOCUS       sequence-259009-        5368 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 5368)
  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 5368)
  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_Zfhx4_enh_12_Citrine
FEATURES             Location/Qualifiers
     source          1..5368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     138..159
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    427..475
                     /note="synthetic polyadenylation signal"
     misc_feature    489..580
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     529..548
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        1615..1760
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1838..1970
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1953..1972
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2024..2033
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2030..2032,2033..2035,2036..2746)
                     /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(2075..2096)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2336..2355)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2683..2704
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2918..3039
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2929..2948)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2983..3002
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3287..3304)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3458..4046)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3538..3557)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4217..5008,5009..5077))
                     /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     4840..4859
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5078..5182)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5209..5368,1..296)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5296..5315)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
       61 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
      121 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
      181 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
      241 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttgcca
      301 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
      361 acgccagccc aagctaccat gataagtaag taatattaag gtacgggagg tacttggagc
      421 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
      481 atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag
      541 gctgtcccca gtgcaagtgc aggtgccaga acatttctct atcgataggt accgagctcg
      601 agttgccagg gctacaaaac ccagttctgc actgaacttc agaggaaatc tgatttttct
      661 tcttttgtaa cacttccatt atgccagctt cggctttgaa taaaatcaag gctttttaaa
      721 actggccaga attaagttac ttcagaaata taacatgggg caaaatcaca catgaaaact
      781 tggctgaact cctcaaactt tactacagcc acaaacaaac ctgactgtta aaatgtaaga
      841 aaaacaagtc tgccattagg taaaaataaa caaggctaaa ggcttcagag tactatgaca
      901 tttcacaccg gctacactgt gaagtccctg tctagcactg acaaaacctg agccccagaa
      961 tcagtgtctt cagctcttgt tgcccctcaa acagaggcat ctttcttggc acactctgcc
     1021 tcacagataa tgttgtgtga gaagcttccc ttaacttcat gggagggatc tatttccatc
     1081 cttgggaagc atccatttac aggacagact gcaagctgag actgggtcct gctgcatccc
     1141 aggaatggag gcagggagga gcccttgtac aattggattc tggccttaca caataagaag
     1201 caattaactg catagctttc tggctcaggc agaaacccaa atgcaagtgc atgccttttc
     1261 tcttccagtc tcagggagca aaaagcagca aattcatata tgctgttacc tgccttctcc
     1321 tagtagccag agcacaatat gagcaatcac atctgaaaga ttgtactgta agctgctgct
     1381 tctccactga ggccagaatc ccatgatata ggaaggtaac agccattagt cttagcactg
     1441 ctaattgctg tttatcaatt gagagtggtg ctggatgtgt gggttttctg ttggctagcc
     1501 cgggctcgag atctgcgatc taagtaagct tgctgcttca tccccgtggc ccgttgctcg
     1561 cgtttgctgg cggtgtcccc ggaagaaata tatttgcatg tctttagttc tatgatgaca
     1621 caaaccccgc ccagcgtctt gtcattggcg aattcgaaca cgcagatgca gtcggggcgg
     1681 cgcggtccca ggtccacttc gcatattaag gtgacgcgtg tggcctcgaa caccgagcga
     1741 ccctgcagcg acccgcttaa cagcgtcaac agcgtgccgc agatcccaag cttgcggccg
     1801 cttaagctgc agaagttggt cgtgaggcac tgggcaggta agtatcaagg ttacaagaca
     1861 ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg
     1921 ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag gtgtccactc
     1981 ccaggttcaa tacagctctt aagcggccgc aagctagctt gccgccacca tggtgagcaa
     2041 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     2101 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     2161 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     2221 cttcggctac ggcctgatgt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     2281 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     2341 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     2401 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     2461 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     2521 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     2581 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     2641 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     2701 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtgag cctctagact
     2761 ccctcagcgg cgcgcctggt gttcctcgag atcggagtcc taaatcgact gccttagcgg
     2821 taggacagta gatcagtcag cgacgtgacg aaccctcgca ggatcaagat cccacgtgtc
     2881 tagagtcggg gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     2941 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     3001 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     3061 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
     3121 aaatgtggta aaatcgataa ggatccgtcg accgatgccc ttgagagcct tcaacccagt
     3181 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
     3241 tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg ctcactgact
     3301 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3361 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3421 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3481 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3541 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3601 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3661 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3721 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3781 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3841 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3901 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3961 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4021 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4081 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4141 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4201 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4261 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4321 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4381 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4441 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4501 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4561 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4621 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4681 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4741 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4801 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4861 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4921 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4981 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5041 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5101 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5161 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta
     5221 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5281 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5341 ttccccgtca agctctaaat cgggggct
//
