LOCUS       sequence-258443-        5535 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 5535)
  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 5535)
  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_Sox9_enh_283_Citrine
FEATURES             Location/Qualifiers
     source          1..5535
                     /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        1307..1452
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1530..1662
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1645..1664
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1716..1725
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1722..1724,1725..1727,1728..2438)
                     /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(1767..1788)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2028..2047)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2375..2396
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2610..2731
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2621..2640)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2675..2694
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2979..2996)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3150..3738)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3230..3249)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3909..4700,4701..4769))
                     /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     4532..4551
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4770..4874)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4901..5356
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4988..5007)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5198..5219
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5487..5535
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
       61 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
      121 gctcgagttg ccaggctcga gagcacaatg aggggggaag gaagaaaaaa aaaaattaga
      181 aaaaaagcca atcaagcagg gatcagctgt gggctgggct gctgcaggtc tgcctgtccc
      241 agaggggggt gcggggagac atattgggcc gtgtgggaaa cccacaggct cagcttctca
      301 cacattgttc tccccggctc cccccacccc cagccaaagc cgttaatgcc acaaaagata
      361 ctttgcccat tgtgccgcgg acagctgaga ggatagagtt agccttacac agctttcctg
      421 cctccctctc ccgctggaga ggggaagagg ggacagagca atcaaaatgg agagtttgat
      481 gctcaccata caaaagaatt aaatccactt aggctccttt ctcgctaatc taggactgtg
      541 gggggttaaa catggttaca ttttcattag gtactaatag caaaagggca gctaatggta
      601 gcacagacca ttgcattgtc atgcagatgg ctcccaaaag ggctacccag catcgcccac
      661 cggccgcagc acgaaggcat gctcggagtg ggaaattgtg tcactcggca ttgttggggg
      721 ctgaattcca tcaatgggaa agtcatgttt cctcggcgga aaggaaacaa gttaaagggc
      781 tcggggagag gaggaggagg tggggagggg agcggcggcg tgcccagctc tgcacctggg
      841 ctgcagggac ccctgtgcgg cacgatgcgg tggcacggag cagtgaccgc ccccggggat
      901 gagctgcagg ggttcggtgg tctctgggca cagtgagtta atggccataa ggtgttagct
      961 ctaaaatacc aactcttgct gtccaggcct ggaaggatgc tccagcccct tccatttcat
     1021 tccagggctg tgatgagcaa cgcatcagtg ataaggtcat gtgaatttgg tctttttggt
     1081 cccattgccc ttacatattt tactctctgt cttcttaagc acattatctc tgttcgtctc
     1141 agcctcacag gggctttctc taccctgctt ccactgagcc tgttggctag cccgggctcg
     1201 agatctgcga tctaagtaag cttgctgctt catccccgtg gcccgttgct cgcgtttgct
     1261 ggcggtgtcc ccggaagaaa tatatttgca tgtctttagt tctatgatga cacaaacccc
     1321 gcccagcgtc ttgtcattgg cgaattcgaa cacgcagatg cagtcggggc ggcgcggtcc
     1381 caggtccact tcgcatatta aggtgacgcg tgtggcctcg aacaccgagc gaccctgcag
     1441 cgacccgctt aacagcgtca acagcgtgcc gcagatccca agcttgcggc cgcttaagct
     1501 gcagaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag
     1561 gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac
     1621 ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccac tcccaggttc
     1681 aatacagctc ttaagcggcc gcaagctagc ttgccgccac catggtgagc aagggcgagg
     1741 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     1801 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     1861 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct
     1921 acggcctgat gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1981 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     2041 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2101 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     2161 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     2221 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     2281 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg
     2341 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     2401 ccgccgggat cactctcggc atggacgagc tgtacaagtg agcctctaga ctccctcagc
     2461 ggcgcgcctg gtgttcctcg agatcggagt cctaaatcga ctgccttagc ggtaggacag
     2521 tagatcagtc agcgacgtga cgaaccctcg caggatcaag atcccacgtg tctagagtcg
     2581 gggcggccgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     2641 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     2701 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     2761 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     2821 taaaatcgat aaggatccgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
     2881 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
     2941 aactcgtagg acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc
     3001 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3061 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3121 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3181 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3241 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3301 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3361 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3421 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3481 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3541 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3601 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3661 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3721 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3781 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3841 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3901 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3961 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4021 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4081 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4141 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4201 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4261 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4321 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4381 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4441 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4501 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4561 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4621 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4681 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4741 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4801 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4861 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
     4921 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4981 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     5041 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5101 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5161 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5221 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     5281 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     5341 ttaacgctta caatttgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     5401 gtgcgggcct cttcgctatt acgccagccc aagctaccat gataagtaag taatattaag
     5461 gtacgggagg tacttggagc ggccgcaata aaatatcttt attttcatta catctgtgtg
     5521 ttggtttttt gtgtg
//
