LOCUS       sequence-249937-        5497 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 5497)
  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 5497)
  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_216_Cerulean
FEATURES             Location/Qualifiers
     source          1..5497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..49
                     /note="synthetic polyadenylation signal"
     misc_feature    63..154
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     103..122
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        1325..1470
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1548..1680
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1663..1682
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1734..1743
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1740..1742,1743..1745,1746..2459)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1785..1806)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2046..2065)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2393..2414
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2621..2742
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2632..2651)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2686..2705
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2990..3007)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3161..3749)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3241..3260)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3920..4711,4712..4780))
                     /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     4543..4562
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4781..4885)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4912..5367
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4999..5018)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5209..5230
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta
       61 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
      121 cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat aggtaccgag ctcgagttcc
      181 atggcggtgt ctggcatgga aagagcagct gctcggagag agaataagta ttagaagaaa
      241 gagaaatcct cgtaatcttg ggacgtatat aatctacgtc gccccgaatc ccaaaaggca
      301 gcgggcccgt ctctctgatt tatttttatg ctagcctagt ttgcatccca aagaagtaaa
      361 ctgctatttt taaccactgg tttaaagctt aattaaaagc ccctaaactg attgttttga
      421 aaattgattt agaggaattg ccccagggct gagacgctat ctaagccaag ttgcagtccc
      481 agagctcgca gtggtgtctg aatcgttaac cccgcagagc cgggagtcgc cggggctgga
      541 tgacacatgg gctggccgag ctccttgcac cctctgcaaa gaaaggtcct ttgttaggcc
      601 aggctactgg tttgctcggg tggatctccg ctggtttcct ggcgttccga ggtttaattg
      661 aattaaggca gagctcggag atgctggtga caccgggacg gagagctctg cctttcatct
      721 gccggagggg aatgtggctg ggtcggggga cgtggaattt ggggacggga cctcgccatt
      781 cgatcgaagg gagctttacc tgctgaaaag gagatggaag ttggttgaag gcaggggggc
      841 tgcggaaggg aagtggcatt tcggcccctt aattagatgt aaacaagaat agggacgggg
      901 ctccatggtc agacaacttg attgttaagg gattaacacc taagtaaacc aataaaggct
      961 ctgcggggct ggtgagaggg ctgcgggcca ggcgagcccg gcacactaat gggttccccc
     1021 cggcacgacc caccccaaaa cggccctctc cgccccgcca gcgacttttg gagcccggtt
     1081 ttaaggctat aagccaaggc ttgagttatt tcttgctggc cgtgctcctt ggaaagcccc
     1141 cttcgggatg cttcaaggtg aggagcctct cattaccggc acccatcgcc ctaattaagg
     1201 accgctagcc cgggctcgag atctgcgatc taagtaagct tgctgcttca tccccgtggc
     1261 ccgttgctcg cgtttgctgg cggtgtcccc ggaagaaata tatttgcatg tctttagttc
     1321 tatgatgaca caaaccccgc ccagcgtctt gtcattggcg aattcgaaca cgcagatgca
     1381 gtcggggcgg cgcggtccca ggtccacttc gcatattaag gtgacgcgtg tggcctcgaa
     1441 caccgagcga ccctgcagcg acccgcttaa cagcgtcaac agcgtgccgc agatcccaag
     1501 cttgcggccg cttaagctgc agaagttggt cgtgaggcac tgggcaggta agtatcaagg
     1561 ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct
     1621 tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag
     1681 gtgtccactc ccaggttcaa tacagctctt aagcggccgc aagctagctt gccgccacca
     1741 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1801 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     1861 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     1921 tcgtgaccac cctgacctgg ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc
     1981 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     2041 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2101 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2161 agctggagta caacgccatc agcgacaacg tctatatcac cgccgacaag cagaagaacg
     2221 gcatcaaggc caacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     2281 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2341 acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2401 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtgag
     2461 cctctagact ccctcagcgg cgacgttggc tgtagagtcg ccagggcgta tttaactgga
     2521 gggactctcg gttagagttg atacggtccc gtcgcataca gcgcctttgg ccactcacgt
     2581 gtctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt
     2641 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2701 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     2761 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     2821 tacaaatgtg gtaaaatcga taaggatccg tcgaccgatg cccttgagag ccttcaaccc
     2881 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
     2941 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg
     3001 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3061 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3121 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3181 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3241 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3301 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     3361 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3421 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3481 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3541 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3601 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3661 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3721 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3781 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3841 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3901 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3961 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4021 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4081 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4141 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4201 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4261 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4321 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4381 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4441 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4501 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4561 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4621 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4681 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4741 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4801 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4861 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct
     4921 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4981 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     5041 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     5101 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     5161 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     5221 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     5281 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     5341 ttaacaaaat attaacgctt acaatttgcc attcgccatt caggctgcgc aactgttggg
     5401 aagggcgatc ggtgcgggcc tcttcgctat tacgccagcc caagctacca tgataagtaa
     5461 gtaatattaa ggtacgggag gtacttggag cggccgc
//
