LOCUS       sequence-259038-        6204 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 6204)
  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 6204)
  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_TFAP2a_enh_249_Cerulean
FEATURES             Location/Qualifiers
     source          1..6204
                     /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        1978..2123
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2201..2333
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2316..2335
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2387..2396
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2393..2395,2396..2398,2399..3112)
                     /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(2438..2459)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2699..2718)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3046..3067
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3279..3400
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3290..3309)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3344..3363
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3648..3665)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3819..4407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3899..3918)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4578..5369,5370..5438))
                     /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     5201..5220
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5439..5543)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5570..6025
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5657..5676)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5867..5888
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6156..6204
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
       61 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
      121 gctcgagttc catggtgcac ctgatttacc cgagttttga tttttatctt gcctagagct
      181 agaaaaattt taaaaaagag tttggattta aagatatcaa tgaaagagct ttatgagaag
      241 tgtgctcaca tacagaaatg attgaatgag tgaagatctg tataatggaa atgtaatcta
      301 attgaggaaa tccctgctag ttccaagaag actttgacat tttaggtttt agaagtgaga
      361 gataagtttg ggcggatgga cttctatttg aaggtcttgc tctcagtttc aaaaagtgct
      421 gagcacgcac ccttcaagat gcttagaagt ttaggtaccc aaatcagcct tgaatattgt
      481 ggtctacccc atgtacatga tgattcaaag ggaggagaga gaaggagcaa agattcccct
      541 ttccctgcct tcactgtaat gcaaggtaag gttgagtgga agtcactgta aggaaaacat
      601 ataaaggcag aaactgctaa gaattttgag gaccaaaata gggacaataa aaggtgggaa
      661 aagtttaaat agcacaacaa agcctctctt tgagaaacat ccaagatagt ttaagagatt
      721 aggcacagct ctagttatca aaaatgcgga agaatgtgca ggagcagtgg agaagaggct
      781 ttgcaagtaa gcaaggaagc agtaaggact tggatcagcc ccgtgttaat taaagaacac
      841 agttaaaatt gtggcataag atcagattcc tttgtttttg cctggctttc tcctctactc
      901 ttttcttcat gaccatgaat ttattcacat agcaaacctt acagagggat ttacataacg
      961 tcattacttc agggtgtttc ttcactcctg attttcaagt ctttgtcatg ggaatgaaat
     1021 tgctctgctc cagggcccca aggcattggt aacactcttc cacagcttca agctgcagag
     1081 aatagtcctg gctatcttag gaaaagtgta aacagtattt gcctgtcttt gttatgttgg
     1141 gagttttcca cctcaaatcc ctttttgacc ctaccctaag gcttacatac tttgtgagaa
     1201 gggcttccct gctttaattt gaaggagatg gctatcgcgt accaatacca ttcccagaat
     1261 tgggtgaaga atttatgact cagtggggag ttctcattgg ttatcagaga aaagactgag
     1321 taggtgaact ctccaaggtt gctgcaagaa tcccagtatt taatctgaat aagattgcac
     1381 aggatcatct catatgccag tgttggttta ttactttgtc ttaaaatttt ctggtggcag
     1441 tgcttttcta ctactggtaa acaaagacag aatatgataa tcacaaaatc agcataaata
     1501 aaatacatag cattttctgt tggaatggaa aattgttcaa atgcatgatt tgggggaaac
     1561 agaagagttg ctcattgttt tagtttttaa aattcagtgg gttgccccaa aaataatgct
     1621 tcctatttat ttccgtgtaa actacaacag atacaaagag cacaataaca ctatttgata
     1681 gagcaaattg ttagctgcaa aatattcttc gtcaacatag ttgccaccat tagctttgca
     1741 ttttttgcca gtgatgaata agagcctgta ttctgtactt gtaaacatct gctccagtgg
     1801 aggtgatgca tttttgctgt caccaatgct gaaatgcaac acctaccgcc aggaccgcta
     1861 gcccgggctc gagatctgcg atctaagtaa gcttgctgct tcatccccgt ggcccgttgc
     1921 tcgcgtttgc tggcggtgtc cccggaagaa atatatttgc atgtctttag ttctatgatg
     1981 acacaaaccc cgcccagcgt cttgtcattg gcgaattcga acacgcagat gcagtcgggg
     2041 cggcgcggtc ccaggtccac ttcgcatatt aaggtgacgc gtgtggcctc gaacaccgag
     2101 cgaccctgca gcgacccgct taacagcgtc aacagcgtgc cgcagatccc aagcttgcgg
     2161 ccgcttaagc tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca aggttacaag
     2221 acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac tcttgcgttt
     2281 ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca caggtgtcca
     2341 ctcccaggtt caatacagct cttaagcggc cgcaagctag cttgccgcca ccatggtgag
     2401 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     2461 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     2521 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     2581 caccctgacc tggggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     2641 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     2701 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     2761 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     2821 gtacaacgcc atcagcgaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     2881 ggccaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     2941 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     3001 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     3061 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt gagcctctag
     3121 actccctcag cggcgcgcca ctggtacgag aacggtggag tacctttcga gcgccggtaa
     3181 gaagcgtcgc ctgactaaag gttcgggtgc agaccagttc cgacacgcga tctcacgtgt
     3241 ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg
     3301 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3361 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3421 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     3481 caaatgtggt aaaatcgata aggatccgtc gaccgatgcc cttgagagcc ttcaacccag
     3541 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     3601 ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac
     3661 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3721 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3781 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3841 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3901 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3961 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4021 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4081 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4141 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4201 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4261 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4321 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4381 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4441 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4501 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4561 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4621 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4681 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4741 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4801 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4861 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     4921 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     4981 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5041 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5101 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5161 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5221 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5281 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5341 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5401 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5461 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5521 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt
     5581 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     5641 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     5701 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     5761 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     5821 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5881 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     5941 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     6001 aacaaaatat taacgcttac aatttgccat tcgccattca ggctgcgcaa ctgttgggaa
     6061 gggcgatcgg tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt
     6121 aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta ttttcattac
     6181 atctgtgtgt tggttttttg tgtg
//
