LOCUS       sequence-259324-        5491 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 5491)
  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 5491)
  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_TFAP2b_enh_338_Citrine
FEATURES             Location/Qualifiers
     source          1..5491
                     /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        1263..1408
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1486..1618
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1601..1620
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1672..1681
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1678..1680,1681..1683,1684..2394)
                     /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(1723..1744)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1984..2003)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2331..2352
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2566..2687
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2577..2596)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2631..2650
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2935..2952)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3106..3694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3186..3205)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3865..4656,4657..4725))
                     /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     4488..4507
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4726..4830)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4857..5312
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4944..4963)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5154..5175
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5443..5491
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
       61 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
      121 gctcgagttg ccaggcaaac acagtgaccg ggagaaggat ttcgaagacc tccgactctg
      181 aggatagaaa agaacgctgc gttaacgtga ctccccaaac tctcttagta acccactccc
      241 ttaaaatgag tgcggaacac gtagaacagc acccaggcac agcgcccgca gcctcagggc
      301 cgcgctccct cagcccggga cgtggccgcc cctcccccag caacggcgcc cacaaaacgg
      361 ccgccacagc gcgattcgag cccccctgga tcctcaatag agcacctacc gttaacctcc
      421 ccctccaaca tcgcctgctc caccgacacc gccatcctcg gcccaagcgc ggctgctcct
      481 gcaacgccag caagcgtggc ccgcctctcc ccccaacagc accactgtgc ctgcgcacgg
      541 aacagcttcc ggggtgaaag gtcacggttg ccgccttctg attggacgga gcggggggcg
      601 ttgggatctc gacgttgtgc ttcgcccttc tgaggcgctg agcccaactg cccttggatg
      661 cccccacccc cctcgaggtg gcggttggag ctccgcgttc aggcctcgtg gccatctttt
      721 tgttctcgtc aaataagcat agccatcccg tgtgcccgga ggggtctgaa agctttgtga
      781 aatctcatct cgtccttagc aagagcaccg agcagggatt tagagacagg atgaggggaa
      841 atggatttgg actggaggag ggcaggttga gactggatgt tgggaagagg ttctttgctg
      901 tgagacactg gaacaggttg cccagtgagg ctgtggatgt cccctccctg ggagcactcc
      961 aggccaggca ggatggggct gtgagcgacc tgggctagag agaggagtcc ctgcctatag
     1021 cagggggttg gaactggatg aacttaaagg ccccttctca cccaaaccac tcttattcaa
     1081 tgattgaagt taagcgcgtc ccagcccaag ccagtgcagg tgttggcagc tattgctgtt
     1141 ggctagcccg ggctcgagat ctgcgatcta agtaagcttg ctgcttcatc cccgtggccc
     1201 gttgctcgcg tttgctggcg gtgtccccgg aagaaatata tttgcatgtc tttagttcta
     1261 tgatgacaca aaccccgccc agcgtcttgt cattggcgaa ttcgaacacg cagatgcagt
     1321 cggggcggcg cggtcccagg tccacttcgc atattaaggt gacgcgtgtg gcctcgaaca
     1381 ccgagcgacc ctgcagcgac ccgcttaaca gcgtcaacag cgtgccgcag atcccaagct
     1441 tgcggccgct taagctgcag aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt
     1501 acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag aagactcttg
     1561 cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt
     1621 gtccactccc aggttcaata cagctcttaa gcggccgcaa gctagcttgc cgccaccatg
     1681 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     1741 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     1801 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1861 gtgaccacct tcggctacgg cctgatgtgc ttcgcccgct accccgacca catgaagcag
     1921 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     1981 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     2041 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     2101 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     2161 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     2221 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     2281 ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     2341 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtgagcc
     2401 tctagactcc ctcagcggcg cgcctggtgt tcctcgagat cggagtccta aatcgactgc
     2461 cttagcggta ggacagtaga tcagtcagcg acgtgacgaa ccctcgcagg atcaagatcc
     2521 cacgtgtcta gagtcggggc ggccggccgc ttcgagcaga catgataaga tacattgatg
     2581 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     2641 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     2701 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     2761 acctctacaa atgtggtaaa atcgataagg atccgtcgac cgatgccctt gagagccttc
     2821 aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact
     2881 gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctcttccg cttcctcgct
     2941 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3001 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3061 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3121 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3181 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3241 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3301 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3361 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3421 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3481 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3541 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3601 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3661 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3721 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3781 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3841 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3901 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3961 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4021 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4081 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4141 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4201 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4261 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4321 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4381 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4441 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4501 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4561 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4621 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4681 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4741 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4801 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc
     4861 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4921 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4981 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5041 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     5101 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     5161 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     5221 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5281 gaattttaac aaaatattaa cgcttacaat ttgccattcg ccattcaggc tgcgcaactg
     5341 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagcccaagc taccatgata
     5401 agtaagtaat attaaggtac gggaggtact tggagcggcc gcaataaaat atctttattt
     5461 tcattacatc tgtgtgttgg ttttttgtgt g
//
