LOCUS       sequence-258732-        5311 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 5311)
  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 5311)
  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_345_Citrine
FEATURES             Location/Qualifiers
     source          1..5311
                     /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        1081..1226
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1304..1436
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1419..1438
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1490..1499
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1496..1498,1499..1501,1502..2212)
                     /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(1541..1562)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1802..1821)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2149..2170
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2386..2507
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2397..2416)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2451..2470
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2755..2772)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2926..3514)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3006..3025)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3685..4476,4477..4545))
                     /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     4308..4327
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4546..4650)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4677..5132
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4764..4783)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4974..4995
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5263..5311
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
       61 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
      121 gctcgagttg ccaggagaac cctcacttga atgccaattt tcaatgttag agatgctatt
      181 gtaattagag aggaatctga cccacaggga tcactttgga aactgcattt gtgtaaattc
      241 tgctgtctgg ggttttggat ccagctcacc acatccagcc cttcttcagc cccttggtgg
      301 aaggagaaca ctgcattcct cccactccat tccccgaggc caggacttcc attttagccc
      361 caaggcaggt gaataattga taaacccatg ctgcatctgc ccctgataga ggcggcagcc
      421 tgcagccagc agtccggctt tgatcggctg ctgcctaaag ccgagaaatc cctttgtagg
      481 cactgaacag ggaaggcatc ccctgtcaaa gtgactggtt acaaaccttc cctggccatg
      541 ggaggccttt tgatgcaggg agcgttgata attagggcct aattgagtga ttaacagccc
      601 cagcagctca tgcctctaag ggtacacgag gtccctacaa agccttggtt tctccagccc
      661 tgccttccct ggcctcggct ccaggctggg tttggtctta gatcagatct tcaccagaaa
      721 tgtgcggagc aagcaacatc acagaaagtg gttacatttt ctcacgagtg ctataatcac
      781 tttctctaaa cctctattga taataactgt tgcaggaata ggctcccatt tgccaacaca
      841 catggtctct gcaggcaatg aaagtctgtc tagatatact gaacttaatt aactgagttt
      901 gaagagccta ttaaggcaat attaaaagaa gcctatctca gcaggccctc acactgttgg
      961 ctagcccggg ctcgagatct gcgatctaag taagcttgct gcttcatccc cgtggcccgt
     1021 tgctcgcgtt tgctggcggt gtccccggaa gaaatatatt tgcatgtctt tagttctatg
     1081 atgacacaaa ccccgcccag cgtcttgtca ttggcgaatt cgaacacgca gatgcagtcg
     1141 gggcggcgcg gtcccaggtc cacttcgcat attaaggtga cgcgtgtggc ctcgaacacc
     1201 gagcgaccct gcagcgaccc gcttaacagc gtcaacagcg tgccgcagat cccaagcttg
     1261 cggccgctta agctgcagaa gttggtcgtg aggcactggg caggtaagta tcaaggttac
     1321 aagacaggtt taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg
     1381 tttctgatag gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt
     1441 ccactcccag gttcaataca gctcttaagc ggccgcaagc tagcttgccg ccaccatggt
     1501 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1561 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1621 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1681 gaccaccttc ggctacggcc tgatgtgctt cgcccgctac cccgaccaca tgaagcagca
     1741 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1801 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1861 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1921 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     1981 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     2041 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     2101 gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     2161 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtgagcctc
     2221 tagactccct cagcggcgcg ccgttgcgtt ccaaacgtcg tggagctccc tgtcaagaca
     2281 cgaaccgcta acgccacctt agtccgtgaa gcatgatacg cgacataccc caggagatct
     2341 cacgtgtcta gagtcggggc ggccggccgc ttcgagcaga catgataaga tacattgatg
     2401 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     2461 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     2521 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     2581 acctctacaa atgtggtaaa atcgataagg atccgtcgac cgatgccctt gagagccttc
     2641 aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact
     2701 gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctcttccg cttcctcgct
     2761 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2821 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2881 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2941 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3001 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3061 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3121 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3181 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3241 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3301 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3361 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3421 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3481 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3541 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3601 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3661 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3721 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3781 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3841 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3901 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3961 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4021 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4081 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4141 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4201 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4261 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4321 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4381 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4441 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4501 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4561 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4621 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc
     4681 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4741 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4801 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     4861 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     4921 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4981 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     5041 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5101 gaattttaac aaaatattaa cgcttacaat ttgccattcg ccattcaggc tgcgcaactg
     5161 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagcccaagc taccatgata
     5221 agtaagtaat attaaggtac gggaggtact tggagcggcc gcaataaaat atctttattt
     5281 tcattacatc tgtgtgttgg ttttttgtgt g
//
