LOCUS       sequence-258510-        8730 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 8730)
  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 8730)
  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_Pax7_enh_199_Citrine
FEATURES             Location/Qualifiers
     source          1..8730
                     /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        4487..4632
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          4710..4842
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4825..4844
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      4896..4905
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4902..4904,4905..4907,4908..5618)
                     /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(4947..4968)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5208..5227)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5555..5576
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5792..5913
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5803..5822)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5857..5876
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6161..6178)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6332..6920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6412..6431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7091..7882,7883..7951))
                     /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     7714..7733
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7952..8056)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8083..8538
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8170..8189)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8380..8401
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    8669..8717
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc
       61 cagtgcaagt gcaggtgcca gaacatttct ctatcgatag gtaccgagct ggttttcggt
      121 ccgggctttt tgctttgttc tgttttaatg tgtcctctgc tgtttaagct ggaagaaata
      181 agcacggggt cctgcctgca gggtgaaaaa aagaccaagt acatctcact cagtcggtag
      241 cagcccgcga gacatatttt tcaccgttct aaaccagcgg accgctaatt tcaacaaagc
      301 ccgcgatttt cgcccactat taggaattta gcctccataa attacaatta acactctccg
      361 cagacaactc aatgggctgg atcgggatgc tgtaacacaa aacggcgttg cccttcgctg
      421 gtgaaattca cagaggaggc cgcttgcttt cccccggggg cagcgctcct ccccacgcat
      481 cggagcaggc aaaccatttt cataggcgct accaaaattt gtgatgacac agggagtgaa
      541 ggatgctttc tcctccattt caggctcgct cctttcccac ttcagcccaa ccagggccac
      601 ctcctgcaga cccggtgcag atttccctgg gcggccgact taggagcaat gactgctcac
      661 tcaagcaagc gccagctggc attatggaga tttaccccca gtgacatctg aattgagctt
      721 tgagttaaac gccctcgtat tcggtccact cgacttcaga gggagttttg tctgaatcaa
      781 aaggaatagg gatagagggg aaaaaaaaaa agaaagattt taggacttag ccgttagaga
      841 agccagaatg cccggatttc cctctcttgt ggcatttggg cactgaggaa agcgccggga
      901 cttaaactga actatcaact cttgaaggaa aaattctccc tggcaagagc agccttgggc
      961 tgatggacgt gtgccggaga tgacctacta agtctttttc tttctcttgc tgttaagttt
     1021 cctcatcccc agctcctggg ttagtcagtt atgctgctaa atctctcctc aattccttat
     1081 gccagcgtct ctctctgcaa ggagacatca aaaccctcct cttcctgcct ctgtaactgt
     1141 aactccctgg accccaccct gcctcttctt cttcattcct ttcactcttt tggaaccaat
     1201 tttgtctcac aaatctcaga tataaccaac ccccacactc acatctccct cacagcaagt
     1261 ggagatgctt ctaatccctt tctaaggagc caaggcaagg tcctcccatc tcttagctcc
     1321 catggagcat tccctaaccc caaagccctt catcctgccc tttccaggcc tcttccagcc
     1381 ccctcagccc tgagcccctt cactgaagcc ttgaaaagga gcacccattg atgccctcag
     1441 aaaatggcca ccgagcaccc aaggcgcctc ccgctactgg caccttctac cacacccagg
     1501 caaggcccat cctgattggc tgcgctgccc acagggggcg ggcctgcggg gcagacagcc
     1561 aatgggagca cagcccctca cctgaggtga gccaggtgga gggaggcggg agctgcttgg
     1621 ctctgcttgc ggaggtgtca gttcagcttt ccttgggctg tgctttattt cttcttcatt
     1681 tcttgttgtg cactccttct ccaccccttt tatcactgct cccgccacag tggtgtaagg
     1741 ggtggtgctg tctgctgact ttctcccccg tggcactcag ctgctgcagc accccacgtc
     1801 atttcagcct ctctttccca gaaacaaacc aaaaaacccc aaacccaaac caaatcgaaa
     1861 acaacagaaa aaaacgcatc caaactgagt ttttttgtgt tgtacgtctc cttggcatca
     1921 ccctggcatc acattacagc acgatgcaaa gtgagtgctc tgaacgtctc catcagtggc
     1981 actcaggtat cacagagaac ctcagagatg cgatttcctt cccaaacttg catatttacc
     2041 ccttttgtta cccaccctcc cccagtttct gctctacaca cctcccctct cacggtcatt
     2101 tggagcacag caaagtatct tgctccttct atgcctcctt tacttctcat ggcctatggg
     2161 gatgtgaggt ggaatgtgag atgcagaatc catccatcac cccctttaga caccacgtac
     2221 agcagaagca agcatgttag gaggtctata tgttactgta tggtctatat tagcctttcc
     2281 catgcttctc agcattgttg tttagatgta tgccttcact aggaagctgc ctgtactgct
     2341 ctatttcaat ttacctgacc atttagactc tgaaagcacc agatcaaagc atttgagcat
     2401 aatttctatc tctaagctta tggaacggga cttagaaact gccatatcag tgcctttttg
     2461 tttgcacgtc aacaggcaaa atgttaaaac aactgtgtaa gtgcttctgc acagcaagcc
     2521 aaggaggggt gaagatggga aaggggattc tggcctcgtc tcatgaatat aaaagcattt
     2581 ctaataagta acagttattc atggtcctgt tcggatagct tctaattcct cccaggcttt
     2641 cagttctcag agcaggatct tgcccgcaag taatgaagag tttgtagggt ttttgggaga
     2701 acttttccag cctaggcctt cagactcaaa acctatgtcc tttcagatgc tgcctacaat
     2761 ctgaataact attttttaat gaaggaagtg acatttttta attcctattt tggcacaagt
     2821 caaccataag tgaattggca ctaggaaatc accattgctt ccactggctg atgggtgcag
     2881 gagataatga cagggtagtt tgcatctcca ttgggaagca tctgccacgg ggaactgtca
     2941 gaggaaggac tgtggatatt aaatgaacca tttatctctt ttgttataaa aactcctatt
     3001 tgaatgtgtc agcagagggt gacattgtgc aggtttggga tataatcttg agtgccatca
     3061 acaacaattt ggaagtctgt gcgcttagaa tggatcccag tttaaagaag gtaggttaag
     3121 tgttatgaaa atcgatgaca aaatataagg aattattttg atgttgcata actctgaaat
     3181 acaaagatag atattgaaaa tctaaatcta atttctgagc tgaaaaattc ttatttttag
     3241 aaatcctgat acttcattta tttattttta acatcaaaga tattcacctc ctctagtttg
     3301 gacgtggctt tggatggaaa cctcagtttt ggggagggaa tctcttgtgt cttcgtatga
     3361 aattgatcca agtgaaactc tgcaggaaaa tgaagtaata tagatggtag atatagatgt
     3421 agtgaagaaa aatgagtgcc gttcctacca ctcctccaag gttggctgtg gatggctatg
     3481 aactaaatga acttaggcag catgcaaacg tttgtaaccc gtctcccaaa gagtttgtgt
     3541 gctggattgg gttgatatca gcctatggag caggtagtat gcatttccat ggctctgtgc
     3601 agcacctagc acgacaaagc tcccacggag tcagcacgag gcagatatct ttcataatca
     3661 atgagggaaa gattgttctt aggtacaaag ggagcgctca gtctccgctg ggttgagttg
     3721 ggtgcctcca cctaatacta agtaggtgca aggcgtgtgc tgctctttga aagtctttcc
     3781 cagaaatctc agcatctcgc gcagatcgca gagacacagt gtgcaaagcg tgcctttaaa
     3841 agacccagtc atggcccgcg cgacgaagtt caaggcttgc tggaacttct tgagttgtgt
     3901 ctgtgtgtgt gtcagctgtc cccgagccca ataccgaacg ggggacaata cagacagata
     3961 agataatgga tgagaaggga gtggaaaatc tagcctgtat tctggtggca tttccaatag
     4021 cgatagcgct ggcagcattg gaccgaaaag cgtatccagc acaaagggct ggtaagtcct
     4081 ctccagctcc cactgtgacg ctaaagccgg ttgcaaatat ccaacgctgg ggcccctcgc
     4141 agtggggagt gggacggtgg ctacgtgacc cggggcagcc ctgccattcc cagaatgggg
     4201 ctcagctccc gccgcttgaa tatcccggtg gggtttctcc cccaaaccac agcaagccca
     4261 tggttaaaat taatatccag catccagctc atcggcgtgc attttaccca agtttcactt
     4321 ctaggcttgt tgcttgctct tctgatggcc catcgtctcc tgttggctag cccgggctcg
     4381 agatctgcga tctaagtaag cttgctgctt catccccgtg gcccgttgct cgcgtttgct
     4441 ggcggtgtcc ccggaagaaa tatatttgca tgtctttagt tctatgatga cacaaacccc
     4501 gcccagcgtc ttgtcattgg cgaattcgaa cacgcagatg cagtcggggc ggcgcggtcc
     4561 caggtccact tcgcatatta aggtgacgcg tgtggcctcg aacaccgagc gaccctgcag
     4621 cgacccgctt aacagcgtca acagcgtgcc gcagatccca agcttgcggc cgcttaagct
     4681 gcagaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag
     4741 gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac
     4801 ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccac tcccaggttc
     4861 aatacagctc ttaagcggcc gcaagctagc ttgccgccac catggtgagc aagggcgagg
     4921 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     4981 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     5041 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct
     5101 acggcctgat gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     5161 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     5221 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     5281 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     5341 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     5401 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     5461 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg
     5521 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     5581 ccgccgggat cactctcggc atggacgagc tgtacaagtg agcctctaga ctccctcagc
     5641 ggcgcgccac gaagctggta gagtgctggc cttgtgccaa gttgcgggag tacccggatc
     5701 tgcgtttaca tacccaccac tcaccgtctt aaagcgggac agatctcacg tgtctagagt
     5761 cggggcggcc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     5821 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     5881 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     5941 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     6001 ggtaaaatcg ataaggatcc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc
     6061 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
     6121 gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc
     6181 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6241 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6301 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6361 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6421 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6481 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6541 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6601 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6661 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6721 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6781 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6841 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6901 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6961 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7021 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7081 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7141 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7201 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7261 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7321 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7381 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7441 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7501 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7561 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7621 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7681 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7741 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7801 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7861 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7921 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7981 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8041 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
     8101 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     8161 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     8221 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     8281 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     8341 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     8401 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     8461 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     8521 tattaacgct tacaatttgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     8581 cggtgcgggc ctcttcgcta ttacgccagc ccaagctacc atgataagta agtaatatta
     8641 aggtacggga ggtacttgga gcggccgcaa taaaatatct ttattttcat tacatctgtg
     8701 tgttggtttt ttgtgtgaat cgatagtact
//
