LOCUS       sequence-259325-        5814 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 5814)
  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 5814)
  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_Pax6_enh_2_Cerulean
FEATURES             Location/Qualifiers
     source          1..5814
                     /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        1585..1730
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1808..1940
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1923..1942
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1994..2003
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2000..2002,2003..2005,2006..2719)
                     /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(2045..2066)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2306..2325)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2653..2674
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2889..3010
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2900..2919)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2954..2973
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3258..3275)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3429..4017)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3509..3528)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4188..4979,4980..5048))
                     /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     4811..4830
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5049..5153)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5180..5635
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5267..5286)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5477..5498
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5766..5814
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
       61 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
      121 gctcgagttc catggggcag ggaaagaaaa cctcttgcta tttttctcat tggtattcac
      181 acggaggggg aaggtggtcc ttctgtgttt ctgtcatgaa taattttgag tgcggggtgg
      241 aaattccaca gtgtatgtag ggcatgaaag tctagggact tggattcact gcaggaccgt
      301 gttgtgcttc aacccaagtt cttgcatttg caatcacaga gtgtgagcgt gtgggggtgg
      361 aattgtatcc acagatgttt agatggaggt gtaaacctcc tcaaatcatt tacagaggag
      421 gtgggagagc agctttgttt ttcagagcag gcacgctgtg gctgaatact tatcaggggc
      481 agtgcattgt cactgcgatg aagaaatagc agtgaggaag ctggtaacag ttatcccttg
      541 tttggttata agcaagagaa aaacctacgt atgtgctaat ctgcatatac actaatcctt
      601 ttgttgtctt agaggtagct agaatctctt gttaaaaaca ctttaaaaaa gaaagcaagg
      661 attggttttg cttttctgga caggagtgaa gctggtgctg aatttttggt tctgtggcta
      721 aatagtggga acgtgagtgc tggtcctgca gtccatgctg aagttactga ttttggaatc
      781 cttatctgca gcagccatta gaagaggaag caccttgatt aaagagagtg gagatttgca
      841 aagagttgga agggctgtgc aggcagctgg agtctatggc cagggccttg tttgaaagaa
      901 aaatattctc aaagcaagct gagccagcac tgggccagca ctgaattgtc ttcatcacaa
      961 aatcatagtc tctttcaaac agaagcacgt ggaggggtgg agacaaaatt attggagtca
     1021 ctgagcaact ttagtttgat gcgagaggca gcagatgtgc agagtgttaa ctggagatgc
     1081 aaaacactcc tgatgtgtat cccctgatat ctgtacagac gtgttagccg ctcatacagc
     1141 tctgcctaca aatccaggtt gtacagttta atctttttca ctcttatttc tctgtgaatt
     1201 agcatttcag tgctttgcct attggttgca ccgatagcac tgttcagctg tgccagcctg
     1261 ccaaggatgg ctagtgatga cagtgggagc ctaattagtg gaagcctaat ccatgtcgag
     1321 gtaatccagt taactgttaa ctgttaaact ttgcagctgt gctgttggct acagtgatcc
     1381 ctgagttgca ctgagttgtg ttggtttttt gtttgtttgt tttttcaatc ctctaggatg
     1441 cctacctaag gaaaacaagg accgctagcc cgggctcgag atctgcgatc taagtaagct
     1501 tgctgcttca tccccgtggc ccgttgctcg cgtttgctgg cggtgtcccc ggaagaaata
     1561 tatttgcatg tctttagttc tatgatgaca caaaccccgc ccagcgtctt gtcattggcg
     1621 aattcgaaca cgcagatgca gtcggggcgg cgcggtccca ggtccacttc gcatattaag
     1681 gtgacgcgtg tggcctcgaa caccgagcga ccctgcagcg acccgcttaa cagcgtcaac
     1741 agcgtgccgc agatcccaag cttgcggccg cttaagctgc agaagttggt cgtgaggcac
     1801 tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt
     1861 gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca
     1921 ctttgccttt ctctccacag gtgtccactc ccaggttcaa tacagctctt aagcggccgc
     1981 aagctagctt gccgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     2041 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     2101 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     2161 gcccgtgccc tggcccaccc tcgtgaccac cctgacctgg ggcgtgcagt gcttcgcccg
     2221 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2281 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2341 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2401 cggcaacatc ctggggcaca agctggagta caacgccatc agcgacaacg tctatatcac
     2461 cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga
     2521 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2581 gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga
     2641 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2701 ggacgagctg tacaagtgag cctctagact ccctcagcgg cgcgccagga catgagtact
     2761 tcccgagaga gcgatgtcaa ccgaatcctg acgggacttc gtaattcgga ccaggtgttc
     2821 ggagcttgtg aggagataga tctcacgtgt ctagagtcgg ggcggccggc cgcttcgagc
     2881 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2941 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3001 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     3061 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccgtc
     3121 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     3181 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     3241 cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3301 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3361 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3421 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3481 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3541 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3601 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3661 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3721 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3781 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3841 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3901 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3961 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4021 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4081 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4141 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4201 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4261 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4321 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4381 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4441 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4501 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4561 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4621 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4681 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4741 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4801 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4861 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4921 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4981 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5041 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5101 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5161 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     5221 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     5281 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     5341 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     5401 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5461 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     5521 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     5581 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttgccat
     5641 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     5701 cgccagccca agctaccatg ataagtaagt aatattaagg tacgggaggt acttggagcg
     5761 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtg
//
