LOCUS       sequence-258442-        4892 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 4892)
  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 4892)
  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_5_Citrine
FEATURES             Location/Qualifiers
     source          1..4892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        559..704
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          782..914
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     897..916
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      968..977
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(974..976,977..979,980..1690)
                     /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(1019..1040)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1280..1299)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1627..1648
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1862..1983
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1873..1892)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1927..1946
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2231..2248)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2402..2990)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2482..2501)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3161..3952,3953..4021))
                     /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     3784..3803
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4022..4126)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4153..4608
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4240..4259)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4450..4471
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4739..4787
                     /note="synthetic polyadenylation signal"
     misc_feature    4801..4892
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4841..4860
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 atcgataggt accgagctcg agttgccagg aaagttttgg gcctgcattg tggttatccg
       61 gggaaacctc tgcaacagca gtcccatcca gcttttagca tccatgggta attctaagag
      121 cagcccttgt tttttcccca aagacttagc ctgcctgcaa gcacaaatat cttagctggc
      181 accattcttc ccgcagtgtc actgaatgac cggattcgtt gccccaggca tcgcagacac
      241 acatccagcg agggtcatcc tagcaagaac aatgctgcca ttgcctttga tttggggaaa
      301 ttgcagcctg ctgggttctg cacagttttg tacttgagat atttttttgt cagcttgcag
      361 ccatctcctg cccaaaccct gtaattgtcg gaggttcggt tgtgctgctg cctttggctg
      421 cagttatctc actgttggct agcccgggct cgagatctgc gatctaagta agcttgctgc
      481 ttcatccccg tggcccgttg ctcgcgtttg ctggcggtgt ccccggaaga aatatatttg
      541 catgtcttta gttctatgat gacacaaacc ccgcccagcg tcttgtcatt ggcgaattcg
      601 aacacgcaga tgcagtcggg gcggcgcggt cccaggtcca cttcgcatat taaggtgacg
      661 cgtgtggcct cgaacaccga gcgaccctgc agcgacccgc ttaacagcgt caacagcgtg
      721 ccgcagatcc caagcttgcg gccgcttaag ctgcagaagt tggtcgtgag gcactgggca
      781 ggtaagtatc aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag
      841 acagagaaga ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc
      901 ctttctctcc acaggtgtcc actcccaggt tcaatacagc tcttaagcgg ccgcaagcta
      961 gcttgccgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1021 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1081 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1141 gccctggccc accctcgtga ccaccttcgg ctacggcctg atgtgcttcg cccgctaccc
     1201 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     1261 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     1321 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     1381 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     1441 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     1501 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1561 gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg
     1621 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1681 gctgtacaag tgagcctcta gactccctca gcggcgcgcc tggtgttcct cgagatcgga
     1741 gtcctaaatc gactgcctta gcggtaggac agtagatcag tcagcgacgt gacgaaccct
     1801 cgcaggatca agatcccacg tgtctagagt cggggcggcc ggccgcttcg agcagacatg
     1861 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     1921 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     1981 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     2041 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccgat
     2101 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactatcgt
     2161 cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct
     2221 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2281 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2341 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2401 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2461 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2521 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2581 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2641 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2701 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2761 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2821 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2881 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2941 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3001 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3061 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3121 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3181 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3241 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3301 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3361 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3421 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3481 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3541 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3601 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3661 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3721 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3781 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3841 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3901 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3961 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4021 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4081 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4141 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4201 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4261 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4321 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4381 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4441 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4501 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4561 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttgc cattcgccat
     4621 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     4681 ccaagctacc atgataagta agtaatatta aggtacggga ggtacttgga gcggccgcaa
     4741 taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact
     4801 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
     4861 cagtgcaagt gcaggtgcca gaacatttct ct
//
