LOCUS       sequence-258288-        4956 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 4956)
  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 4956)
  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_Snai2_enh_13_Citrine
FEATURES             Location/Qualifiers
     source          1..4956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          78..210
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     193..212
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      264..273
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(270..272,273..275,276..986)
                     /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(315..336)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(576..595)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     923..944
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1158..1279
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1169..1188)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1223..1242
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1527..1544)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1698..2286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1778..1797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2457..3248,3249..3317))
                     /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     3080..3099
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3318..3422)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3449..3904
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3536..3555)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3746..3767
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4035..4083
                     /note="synthetic polyadenylation signal"
     misc_feature    4097..4188
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4137..4156
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        4811..4956
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
ORIGIN
        1 cagcgtcaac agcgtgccgc agatcccaag cttgcggccg cttaagctgc agaagttggt
       61 cgtgaggcac tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga
      121 aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta
      181 ctgacatcca ctttgccttt ctctccacag gtgtccactc ccaggttcaa tacagctctt
      241 aagcggccgc aagctagctt gccgccacca tggtgagcaa gggcgaggag ctgttcaccg
      301 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
      361 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
      421 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgatgt
      481 gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
      541 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
      601 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
      661 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
      721 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
      781 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
      841 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag
      901 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
      961 ctctcggcat ggacgagctg tacaagtgag cctctagact ccctcagcgg cgcgcctggt
     1021 gttcctcgag atcggagtcc taaatcgact gccttagcgg taggacagta gatcagtcag
     1081 cgacgtgacg aaccctcgca ggatcaagat cccacgtgtc tagagtcggg gcggccggcc
     1141 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     1201 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     1261 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     1321 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa
     1381 ggatccgtcg accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc
     1441 ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac
     1501 aggtgccggc agcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1561 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     1621 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     1681 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     1741 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     1801 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     1861 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     1921 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     1981 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2041 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2101 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2161 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2221 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2281 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2341 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2401 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2461 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2521 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     2581 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     2641 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     2701 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     2761 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     2821 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     2881 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     2941 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3001 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3061 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3121 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3181 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3241 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3301 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3361 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3421 cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg
     3481 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3541 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3601 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3661 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3721 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3781 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3841 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     3901 atttgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     3961 tcgctattac gccagcccaa gctaccatga taagtaagta atattaaggt acgggaggta
     4021 cttggagcgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt
     4081 gtgaatcgat agtactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag
     4141 caaaataggc tgtccccagt gcaagtgcag gtgccagaac atttctctat cgataggtac
     4201 cgagctcgag ttgccaggtt cgtttcatgc ctcctcggtg tagtctgtct ctctggctga
     4261 ttttcttcct ttctttatct gaggctggca ttttttccac tctcaataat taagccgtct
     4321 cgatgttaga gacatgccta ttttatcaca tgtgtgtctc gtacacagtt gaaaaaggag
     4381 atctccaagg gagaagaagg ctggcgtctc tgctgttgaa acggtttcaa tctggcctga
     4441 atactttgcc tgcagtgctg caacccacaa gggggtctta acctctgaaa cctgctgact
     4501 caagagtaat tggccagcat gatgccactg tgaccatcaa gtggaacaaa taaaattacc
     4561 caccatttca gtgccgtgat tgtcctctcc cactgcatga ttccacgaat tccccggagg
     4621 actgacctgc ttataataaa caaattgttt tctgggctcc ttttccattg cctgtagggg
     4681 agtctgttgg ctagcccggg ctcgagatct gcgatctaag taagcttgct gcttcatccc
     4741 cgtggcccgt tgctcgcgtt tgctggcggt gtccccggaa gaaatatatt tgcatgtctt
     4801 tagttctatg atgacacaaa ccccgcccag cgtcttgtca ttggcgaatt cgaacacgca
     4861 gatgcagtcg gggcggcgcg gtcccaggtc cacttcgcat attaaggtga cgcgtgtggc
     4921 ctcgaacacc gagcgaccct gcagcgaccc gcttaa
//
