LOCUS       sequence-258412-        5306 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 5306)
  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 5306)
  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_372_Citrine
FEATURES             Location/Qualifiers
     source          1..5306
                     /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        1063..1208
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1286..1418
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1401..1420
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1472..1481
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1478..1480,1481..1483,1484..2194)
                     /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(1523..1544)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1784..1803)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2131..2152
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2368..2489
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2379..2398)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2433..2452
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2737..2754)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2908..3496)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2988..3007)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3667..4458,4459..4527))
                     /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     4290..4309
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4528..4632)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4659..5114
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4746..4765)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4956..4977
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5245..5293
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
       61 cagtgcaagt gcaggtgcca gaacatttct ctatcgatag gtaccgagct cgagttgcca
      121 gggcagcagt tgaagcaaac caggacaaca gagctcattt ccagcagcca gccctcgctg
      181 cagcacaatc cagcgtgcct cctggtaatg aaattagctc tgtcatgtca cgtcaaaatg
      241 aaatccatgt gaaaggaggg tcgcctgctt tccacagccc aggcccaccg atgtacctac
      301 tgtattattt tgtcacactc cagtaacgcc gcaccggggt ctcagtggcc ttcctgcagc
      361 taagcaccat tgaaaacaca tttgtcctgg ctgttctttt cttttacccc tcctctcatc
      421 ttgtgcaaga tgaatcgctg ctggatccag actgactccg agggcaaagc caggtcttga
      481 ggaggttacc tgcctcccac agcttaaaca ttttaattaa ttttaacact aatgcacaaa
      541 tgagagttaa tcactctgct gcagtgctct ctaatttacc gagtttattc attaaaactt
      601 tcctcactga actttgattg aatgtcatgc tttttaagag gcctctcaaa ctgccctaag
      661 gcaaaaaccc atgtctgggg catctgctgc agcactcctt acaatcagct tcacagggaa
      721 gaatgggaga atttggtggg agggggactg ggctggggag ctgaactcgg ttccttaggc
      781 agtgccacaa cagggctgcc tgggagccca gacacgactg caattgccta ctgagcacaa
      841 aaggggaatt ctaacttgca caaactcgag accaaaacag ttcccagccc cacagtataa
      901 agctgtccac gtgatcgttg tgctggcttt tgtcactgtt ggctagcccg ggctcgagat
      961 ctgcgatcta agtaagcttg ctgcttcatc cccgtggccc gttgctcgcg tttgctggcg
     1021 gtgtccccgg aagaaatata tttgcatgtc tttagttcta tgatgacaca aaccccgccc
     1081 agcgtcttgt cattggcgaa ttcgaacacg cagatgcagt cggggcggcg cggtcccagg
     1141 tccacttcgc atattaaggt gacgcgtgtg gcctcgaaca ccgagcgacc ctgcagcgac
     1201 ccgcttaaca gcgtcaacag cgtgccgcag atcccaagct tgcggccgct taagctgcag
     1261 aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt acaagacagg tttaaggaga
     1321 ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat aggcacctat
     1381 tggtcttact gacatccact ttgcctttct ctccacaggt gtccactccc aggttcaata
     1441 cagctcttaa gcggccgcaa gctagcttgc cgccaccatg gtgagcaagg gcgaggagct
     1501 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1561 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     1621 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg
     1681 cctgatgtgc ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1741 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1801 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1861 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1921 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     1981 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     2041 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct
     2101 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     2161 cgggatcact ctcggcatgg acgagctgta caagtgagcc tctagactcc ctcagcggcg
     2221 cgccgttgcg ttccaaacgt cgtggagctc cctgtcaaga cacgaaccgc taacgccacc
     2281 ttagtccgtg aagcatgata cgcgacatac cccaggagat ctcacgtgtc tagagtcggg
     2341 gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     2401 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     2461 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     2521 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     2581 aaatcgataa ggatccgtcg accgatgccc ttgagagcct tcaacccagt cagctccttc
     2641 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     2701 ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2761 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2821 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2881 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2941 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3001 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3061 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     3121 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3181 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3241 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3301 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     3361 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3421 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3481 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3541 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3601 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3661 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3721 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3781 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3841 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3901 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3961 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4021 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4081 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4141 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4201 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4261 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4321 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4381 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4441 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4501 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4561 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4621 aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt
     4681 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4741 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4801 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4861 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4921 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4981 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     5041 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     5101 aacgcttaca atttgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     5161 gcgggcctct tcgctattac gccagcccaa gctaccatga taagtaagta atattaaggt
     5221 acgggaggta cttggagcgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt
     5281 ggttttttgt gtgaatcgat agtact
//
