LOCUS       sequence-258508-        7159 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 7159)
  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 7159)
  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_TFAP2b_enh_36_Citrine
FEATURES             Location/Qualifiers
     source          1..7159
                     /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        2930..3075
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          3153..3285
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3268..3287
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      3339..3348
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3345..3347,3348..3350,3351..4061)
                     /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(3390..3411)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3651..3670)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3998..4019
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4234..4355
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4245..4264)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4299..4318
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4603..4620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4774..5362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4854..4873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5533..6324,6325..6393))
                     /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     6156..6175
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6394..6498)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6525..6980
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6612..6631)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6822..6843
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    7111..7159
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
       61 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
      121 gctcgagttg ccaggtccca gctaaggatg tgggtataat gtatctccca atttctgaga
      181 aagttcaata aactgcaaca catccttcag taaactgaaa aatggcatgg atctattgga
      241 actgaataag ctggggaaac ggaaaaaaaa cgataggcaa gtcacagtag tttgatttat
      301 cacaaggaag cttattttcc actgttcatg gaagaacact gcttgcagta ttctcctggc
      361 ccaggtatgg agctgtgaaa aagaaggaat acttaggaag ccaagggaag cttacagaca
      421 gtgccatcct tatgatgatg ctttcctatc agctagcact gcgacattcg tacatcccaa
      481 aaaactaaaa cctacagaat caatcatttc ctttaaatag ctaggtaaat tcttgtctcc
      541 aagatcctta atgccatata aatatttagc cactggcttc cctgggtaat tatgcctaaa
      601 ctctgccctg aaggcaacca tccctggcac ttaaggctga ctgaaaggca ggagcaaatt
      661 cattgtttag attaaatata tggagacatc atccctaatg tttgggatga tcagctccaa
      721 aaggtacaat gtgggcatat ttcagtgtat tcaaagtaaa aaaatgaggt ccattatttg
      781 ccatagatgc cagctatgcc aaatggatat ttggcattaa aaaagaaaaa agatctgagt
      841 tccagcattg gtaattataa accttttttc aaagaatgtg agcctattca cataccttag
      901 gctgagtaat gaattatatt ttgtggtgtg ctgggaagga ccctgggata tttttcacct
      961 tctgttgagc agggaaaaca tcaggcagac acaaaaggtt tcctgggcag gagtactagg
     1021 ggaccatcag tccctcttat cttcaaagtt acattcagtg tgttttgttt ttcttttagg
     1081 aaagtgcaac tacattccaa ctaaacaaga gagaataaat tctatgtgac ttttgccctg
     1141 tactcttaga gaaagaattt tctcctgtag gatgcctgta tgggatgcag acgaacatta
     1201 aagacctaaa cttatatttt cccttcttca tgcatttgat acaaaggaaa cccttaaaaa
     1261 ttctcatctc ttatattctt tgcccagatg aaaattcccc acacttttgt ccatgtaaac
     1321 acagaattag tctcctgcaa gtttcggtgg tggcagttcc tgaacacagt catattttgc
     1381 agataacagg attagcttgg atgtgggtaa taagttggtg aatctccccc tggccatttc
     1441 tcaacactgt gagagcacta atcctttatg ctttgatgtc ttctagtttt ccatcaggtg
     1501 cagctcggca cttttgtcct ggggcagtca gcgctcttat aagatctcat agtgtcctca
     1561 ttagctgcgc ctatttaccc agccagcccg ttgaacccaa tttatagttt atgtgattaa
     1621 ttcccaaagt acagtaaacc ctagctttaa ggctctctat tatatggctg catctcatat
     1681 aagggcaaac actagctacc gctcttttgt tatatggctt ctctatacat aaaactgtaa
     1741 taatttttct ctcaactaca cattaaacat cattatatat acctcctctc agacatacag
     1801 cctataaaaa tgttgcacgt tggatcataa aggaacatgc atgcacaaga gtcatttttt
     1861 ggttgtttcc tatttttatt ttaaaaatct atgtctttca cattttatat cacaattatt
     1921 gatattgttg agtaagtcta acaaggcaca taatttggaa ctgtttatcc atagaattgc
     1981 aggttacaaa ggcttagtag tcaaagtagc agcctataac cactggcaca attttttttt
     2041 ttttttttta aacatgcagg gtttggagat aaaagactta cgttgtgaat ttttatgcag
     2101 tcatttaagt caactgatga catctgtcag ctctgctgta tttcctccta ttgggatata
     2161 aactatggga gttggatctt tggtaatttg catgagaaag aacattgtat ctttttttaa
     2221 atggatgatc atttccagct atctctcctc aagggttctt tgggagaggc atcctgcagg
     2281 tctctctctt ttctattttt ttctttttct tttttttttt ttttcccccc atccatttag
     2341 tttcagaaac aacactgctt ctcactgctt cttggaagtt tcaattacac tggatgatct
     2401 ccatcacgaa agcaaaagca caaaatcagc aagtctcaga gtattttgag taagatgcag
     2461 aaagtactga gcagagcgga tagcagaaaa gctcttttga aaggggatca ctcagtgtga
     2521 gctcaggtgg gtgaagctgg ttgtttaaac agtaaaaccc aacttgctat acaaaaaggc
     2581 cttatgagaa tcagttacac cacctgaaaa aaacaaaaaa caaaaaacat aactacagat
     2641 tgtagaagac caaattgtta ccttagtgca tactagaagt aatgcaaaca tacagctgtg
     2701 gccaatgaga acctcatttt gacttaaaat gaggtcttat ctgctttatt caatagcctt
     2761 agcaccatct gacaaatggt atgacatacc accttgcccc aactgttggc tagcccgggc
     2821 tcgagatctg cgatctaagt aagcttgctg cttcatcccc gtggcccgtt gctcgcgttt
     2881 gctggcggtg tccccggaag aaatatattt gcatgtcttt agttctatga tgacacaaac
     2941 cccgcccagc gtcttgtcat tggcgaattc gaacacgcag atgcagtcgg ggcggcgcgg
     3001 tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc tcgaacaccg agcgaccctg
     3061 cagcgacccg cttaacagcg tcaacagcgt gccgcagatc ccaagcttgc ggccgcttaa
     3121 gctgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt
     3181 aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg
     3241 cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagg
     3301 ttcaatacag ctcttaagcg gccgcaagct agcttgccgc caccatggtg agcaagggcg
     3361 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     3421 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     3481 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg
     3541 gctacggcct gatgtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     3601 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     3661 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     3721 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3781 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     3841 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     3901 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     3961 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     4021 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtgagcctct agactccctc
     4081 agcggcgcgc ctgtcgaagc taaaagtgct cgagcccaat accatttcgg aatctcgcgt
     4141 ggcgttagta tgccactcga tggtagggga acgcagaatg ccagatctca cgtgtctaga
     4201 gtcggggcgg ccggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     4261 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     4321 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     4381 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     4441 gtggtaaaat cgataaggat ccgtcgaccg atgcccttga gagccttcaa cccagtcagc
     4501 tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc
     4561 atgcaactcg taggacaggt gccggcagcg ctcttccgct tcctcgctca ctgactcgct
     4621 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4681 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4741 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4801 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4861 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4921 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4981 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5041 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5101 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5161 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5221 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5281 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5341 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5401 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5461 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5521 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5581 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5641 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5701 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5761 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5821 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5881 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5941 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6001 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6061 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6121 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6181 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6241 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6301 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6361 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6421 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6481 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg
     6541 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     6601 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     6661 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     6721 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     6781 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     6841 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     6901 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     6961 aatattaacg cttacaattt gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     7021 atcggtgcgg gcctcttcgc tattacgcca gcccaagcta ccatgataag taagtaatat
     7081 taaggtacgg gaggtacttg gagcggccgc aataaaatat ctttattttc attacatctg
     7141 tgtgttggtt ttttgtgtg
//
