LOCUS       sequence-258495-        6563 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 6563)
  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 6563)
  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_366_Citrine
FEATURES             Location/Qualifiers
     source          1..6563
                     /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        2320..2465
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2543..2675
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2658..2677
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2729..2738
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2735..2737,2738..2740,2741..3451)
                     /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(2780..2801)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3041..3060)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3388..3409
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3625..3746
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3636..3655)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3690..3709
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3994..4011)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4165..4753)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4245..4264)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4924..5715,5716..5784))
                     /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     5547..5566
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5785..5889)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5916..6371
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6003..6022)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6213..6234
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6502..6550
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
       61 cagtgcaagt gcaggtgcca gaacatttct ctatcgatag gtaccgagct cgagttgcca
      121 ggtcccctgc ttgccatgta aggtcactgc agacactgta gagctgctcc tgtttgataa
      181 ttgcaaatta gagatccagc ttcaaccaac gcaagatcac ggcagattat cagcaccatg
      241 ggagtgtctc ttagcaggga gaaaaagatc aaataaaaag cccaaactga gccttattag
      301 ctccatactc gcctcccccc atcccctctc ctcgccccca ttcctcctcc cttcccaaac
      361 tgtcatggtt tattttgttt gggtggattt tctttttctt tcctccccct cccagcccct
      421 cgcaaacaga tagttttccc atggagatcc agcacgctct gtgcctatta gcatccttgt
      481 gcataattag agcccagcgc attgccatgc aaagcagcct cggatgacaa cacttcagcc
      541 cggacgacac agaaatgcag cgaggaagtt tctcacactg gttttagcca aggcaaaccg
      601 gactgtgcac gggtttgaga ttttaattat tggatgggtg ctgggctgcc cttacagctc
      661 ggcgaggaaa gctcccagcc acccgcgtgc cccccggcct tcacgttgcc ctctgatact
      721 ttctgctcat tacaatgaat tccaaacccc tctctgaaca gtcactgaat ggttttccac
      781 atcatcaacg ctaaatcagc atctctgcca tgctctgcct ctccgctcat cctagttgcc
      841 aaataggctc aagaaggcaa tgatggttgg aggaaaggtg gagagcttgg cctgggacag
      901 cccagaaggg gatcccacct ccagcatctc tcctgctgca tctcttccac tccacagctc
      961 ccctgccctg gccaacagca tcctgctcag taaaggatgg ctctgcaagc atcccacgta
     1021 ctcaggggag gctcccagtg cccaggagca caccgtgcct gagctcccca agcactcttc
     1081 atgatgccca cagcttctct gaaccccacg ggcatccctc agggcaggat ccttgcagca
     1141 cactgactca cagagtttgc actcccaggg aggtagctgg gaagtttcgg aggtgcccca
     1201 ggtttaatca ttaatcgctg agctccgctc tatggtttcg ccttgggaat ccacctgctc
     1261 tcacaccagt cagaagcagt ggcatttaaa agtggcttcc agagcatcca ggcataaggc
     1321 aggggagagg gtagaggcga gggatggccc cgtccccatc ctatgccatt gctctgaggc
     1381 tgagagagct gaccctgcag agctgcactg gtccaaagag cagcgacttc agccattaac
     1441 gcacctgcga gggccacctc atacatcagt gggtcattaa aggatctgac actcattaac
     1501 agtcccgcag ggcgttatgt gctggtgcag cttcccactg cattatcctt cccgcaggag
     1561 agcagagggg aaaaccgcag gggagagact gcttcaaacc ctgagccctc ttctgctcct
     1621 accctctcta aattgacaag ctcagccaag ggatgactgc tccaccctat taccagtggc
     1681 agagccacag tctgtacatg ctggtgcagg aaccacagca gtaccacttt gctctgccat
     1741 ccccagggct ccgcaccctc acagaggcag cagaacgaat cccagtgctg cagcacccag
     1801 cggatgctgt gggctgggga ggtcatactg gctcccagct gcagcttcca ctgtgtttgc
     1861 tttgcctgcc cccatacaca ccgggctggc agcagggccg acctgcctgc tgacccgtaa
     1921 ccagcatggc cgagccctga ttgcaacctt ctggcagtta atgcgatcct cttgactaca
     1981 agcacagcga gaggagcggc tggtggctct tccctctgct ctgcagggtt ggctggatgg
     2041 gagcctcctg cactgctggc tgagaaaggg cagagcacgg ggtagccggc tccaaatccc
     2101 attgcattgc taaatacagt ccagcaaatg tggcagccct gccccatggg gagatactga
     2161 tagggacgat cctgctgcag gatggaagat ggctgttggc tagcccgggc tcgagatctg
     2221 cgatctaagt aagcttgctg cttcatcccc gtggcccgtt gctcgcgttt gctggcggtg
     2281 tccccggaag aaatatattt gcatgtcttt agttctatga tgacacaaac cccgcccagc
     2341 gtcttgtcat tggcgaattc gaacacgcag atgcagtcgg ggcggcgcgg tcccaggtcc
     2401 acttcgcata ttaaggtgac gcgtgtggcc tcgaacaccg agcgaccctg cagcgacccg
     2461 cttaacagcg tcaacagcgt gccgcagatc ccaagcttgc ggccgcttaa gctgcagaag
     2521 ttggtcgtga ggcactgggc aggtaagtat caaggttaca agacaggttt aaggagacca
     2581 atagaaactg ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg
     2641 tcttactgac atccactttg cctttctctc cacaggtgtc cactcccagg ttcaatacag
     2701 ctcttaagcg gccgcaagct agcttgccgc caccatggtg agcaagggcg aggagctgtt
     2761 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     2821 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     2881 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg gctacggcct
     2941 gatgtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     3001 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     3061 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     3121 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     3181 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     3241 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     3301 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgag
     3361 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     3421 gatcactctc ggcatggacg agctgtacaa gtgagcctct agactccctc agcggcgcgc
     3481 cgttgcgttc caaacgtcgt ggagctccct gtcaagacac gaaccgctaa cgccacctta
     3541 gtccgtgaag catgatacgc gacatacccc aggagatctc acgtgtctag agtcggggcg
     3601 gccggccgct tcgagcagac atgataagat acattgatga gtttggacaa accacaacta
     3661 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     3721 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     3781 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa
     3841 tcgataagga tccgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg
     3901 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc
     3961 gtaggacagg tgccggcagc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4021 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4081 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4141 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4201 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4261 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4321 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4381 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4441 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4501 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4561 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4621 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4681 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4741 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4801 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4861 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4921 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4981 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5041 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5101 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5161 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5221 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5281 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5341 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5401 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5461 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5521 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5581 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5641 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5701 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5761 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5821 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5881 ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag
     5941 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     6001 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     6061 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     6121 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     6181 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     6241 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     6301 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     6361 gcttacaatt tgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     6421 ggcctcttcg ctattacgcc agcccaagct accatgataa gtaagtaata ttaaggtacg
     6481 ggaggtactt ggagcggccg caataaaata tctttatttt cattacatct gtgtgttggt
     6541 tttttgtgtg aatcgatagt act
//
