LOCUS       sequence-258444-        5230 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 5230)
  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 5230)
  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_FoxD3_enh_8_Citrine
FEATURES             Location/Qualifiers
     source          1..5230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..92
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     41..60
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        989..1134
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1212..1344
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1327..1346
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1398..1407
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1404..1406,1407..1409,1410..2120)
                     /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(1449..1470)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1710..1729)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2057..2078
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2292..2413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2303..2322)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2357..2376
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2661..2678)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2832..3420)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2912..2931)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3591..4382,4383..4451))
                     /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     4214..4233
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4452..4556)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4583..5038
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4670..4689)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4880..4901
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5169..5217
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
       61 cagtgcaagt gcaggtgcca gaacatttct ctatcgatag gtaccgagct cgagttgcca
      121 gggtgacacg ttctaaggct cagcagtgct ttggtgggcc gttaagtgtg gtgaaggctg
      181 aaacagcccc tgtaaccgtc ctgcagtaag gaggagtgct catgtgcaca gggctcagct
      241 cctcctcgca caccatccct gttcctgtgg ctgctgggac gagctatggt tcgggttatc
      301 aaacaccacc catttcacag gcgcaaagga aaaaaagccc tacttggtct gtgtgttctg
      361 aatgcatttt gctgcaggag gcagagctgc tgagcgaggt gcagcaggag gcagccctgg
      421 tcctgggggg aagcagctcg aggctttgat cccctcccct ctttgtttgc tcggtgactt
      481 tttgttttct gcattaactt ggaattaatg gtatgtcccc atgctgtgtg gctgctcagt
      541 gtgttccctt cagtcctatg aggtgtttac tgagggcata gaaatacttt gctctgaaat
      601 cagagcttta attcaggagt attagaaaat tgccttagtg atgcataagg ccatctcaca
      661 aacagtggct ttacctcctg aggagaagtg agctgactcc ccctggagct tggtgtcgta
      721 gtgtggctcc tcgtggagat caggtgatgg ctgtgtgctc tgtgaggttt accatcatac
      781 ttccaaaggc tgttcaagaa aaaaagtgat agaaacactc ttggagctgc tgcattctcg
      841 gatagagatg ggagcacaag gctgttggct agcccgggct cgagatctgc gatctaagta
      901 agcttgctgc ttcatccccg tggcccgttg ctcgcgtttg ctggcggtgt ccccggaaga
      961 aatatatttg catgtcttta gttctatgat gacacaaacc ccgcccagcg tcttgtcatt
     1021 ggcgaattcg aacacgcaga tgcagtcggg gcggcgcggt cccaggtcca cttcgcatat
     1081 taaggtgacg cgtgtggcct cgaacaccga gcgaccctgc agcgacccgc ttaacagcgt
     1141 caacagcgtg ccgcagatcc caagcttgcg gccgcttaag ctgcagaagt tggtcgtgag
     1201 gcactgggca ggtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg
     1261 gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca
     1321 tccactttgc ctttctctcc acaggtgtcc actcccaggt tcaatacagc tcttaagcgg
     1381 ccgcaagcta gcttgccgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     1441 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1501 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1561 agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg atgtgcttcg
     1621 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1681 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1741 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1801 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     1861 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     1921 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1981 ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca
     2041 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2101 gcatggacga gctgtacaag tgagcctcta gactccctca gcggcgcgcc tggtgttcct
     2161 cgagatcgga gtcctaaatc gactgcctta gcggtaggac agtagatcag tcagcgacgt
     2221 gacgaaccct cgcaggatca agatcccacg tgtctagagt cggggcggcc ggccgcttcg
     2281 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2341 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2401 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     2461 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc
     2521 gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca
     2581 tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc
     2641 cggcagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2701 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2761 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2821 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2881 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2941 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3001 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3061 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3121 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3181 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3241 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3301 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3361 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3421 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3481 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3541 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3601 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3661 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3721 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3781 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3841 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3901 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3961 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4021 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4081 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4141 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4201 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4261 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4321 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4381 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4441 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4501 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4561 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4621 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4681 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4741 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4801 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4861 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4921 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4981 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttgc
     5041 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     5101 ttacgccagc ccaagctacc atgataagta agtaatatta aggtacggga ggtacttgga
     5161 gcggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat
     5221 cgatagtact
//
