LOCUS       sequence-249926-        5919 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 5919)
  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 5919)
  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_Zfhx4_enh_5_Citrine
FEATURES             Location/Qualifiers
     source          1..5919
                     /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        1740..1885
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1963..2095
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     2078..2097
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2149..2158
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2155..2157,2158..2160,2161..2871)
                     /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(2200..2221)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2461..2480)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2808..2829
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3043..3164
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3054..3073)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3108..3127
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3412..3429)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3583..4171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3663..3682)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4342..5133,5134..5202))
                     /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     4965..4984
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5203..5307)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5334..5789
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5421..5440)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5631..5652
                     /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 ctcgagttgc
      181 caggtgcgag gttcaaggaa aactatttgt tcaactgaaa acaggcagga agaaggatgt
      241 gggaatttag agacagactt tacattcaag actcctcaga atgggaaaaa catagataag
      301 aaaaacttct ctttgaacta ttagagatac tcttaaaaat ataggctgtg tttttccata
      361 acgttatggt tatggaaaat aatattactt ttaattgaag gtatagatct gaaagaatag
      421 aattttatgt tatcatttta aatatatata caaatgactc accctgtttt catcatgaaa
      481 ttccctgaag agtgaagctg taaaacctcc tgagaaattt cctgaagaca taaaggcctt
      541 agaatgttgc ttgaagccac tgccaattca ttactcaaca gaagcaatca gtgtttcatc
      601 tccagaagaa aagacaaaaa aaaaaaaaaa aaaaaaaaaa aattctctta cctcctaggg
      661 acttcctgaa agtaggaaga gaaagagaaa actgagagaa cactactggc tcagtgcttc
      721 cacaaagcac tcctttttcc ttctctcagc tgaatgcaaa tgacctcttc tagtgtggaa
      781 ggtcacctca tcgcccagag aaaagcagaa gaccagaatt cactgtacac tcattgcact
      841 caccctcatt gtacagcaac attggttgtt tggacatcta ggaagggaaa gaggacagat
      901 ctgtatcact ctatcgcctt gcaattcttc gtttccctta ttcacagtaa aatggagaaa
      961 tgcatgctta gaagagatcg ctttttccct tattcctagt aacagagatc aatggcaaat
     1021 aatacagaga ctagcacatt tataggattc aacctcagaa atagttttct gctctactgt
     1081 actttggttt ggctgatttt atgtacaaaa gagaatattc aatattcaaa tggatgccaa
     1141 gcacagtagt atagtgcttt aacctttccc tttgaaatat tttagttcag tgggtggcag
     1201 agaaaaccac aaactgtggc atgactgtca gtctcataaa tgatttgtga ctgtgtaaat
     1261 gcagatgcca cagctgtcat gtaatctgca cacaggctat actgagaatg taactcagac
     1321 ccttctgctc caggaacaca ggccttttcc atttgtactg ttccattagc tgttgaaatg
     1381 gcttaaaact agaacaattt atccctttcc cttttgaact ttagtaaaat aggacagagc
     1441 tctgaaatgg ctttcattta taaaccaaaa ttctcccaat gtgattttct aaagcaaaca
     1501 tttaaatcca gtgatatcta aatagctgtc aacagtatta tagggagctg tagaaaactt
     1561 taaaagtaaa actcagaaat ggatacattt tcctggtttc aagtgtcatt gtctgttggc
     1621 tagcccgggc tcgagatctg cgatctaagt aagcttgctg cttcatcccc gtggcccgtt
     1681 gctcgcgttt gctggcggtg tccccggaag aaatatattt gcatgtcttt agttctatga
     1741 tgacacaaac cccgcccagc gtcttgtcat tggcgaattc gaacacgcag atgcagtcgg
     1801 ggcggcgcgg tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc tcgaacaccg
     1861 agcgaccctg cagcgacccg cttaacagcg tcaacagcgt gccgcagatc ccaagcttgc
     1921 ggccgcttaa gctgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca
     1981 agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt
     2041 ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc
     2101 cactcccagg ttcaatacag ctcttaagcg gccgcaagct agcttgccgc caccatggtg
     2161 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2221 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     2281 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2341 accaccttcg gctacggcct gatgtgcttc gcccgctacc ccgaccacat gaagcagcac
     2401 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     2461 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2521 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2581 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     2641 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     2701 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     2761 agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     2821 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtgagcctct
     2881 agactccctc agcggcgcgc ctggtgttcc tcgagatcgg agtcctaaat cgactgcctt
     2941 agcggtagga cagtagatca gtcagcgacg tgacgaaccc tcgcaggatc aagatctcac
     3001 gtgtctagag tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt
     3061 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3121 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     3181 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     3241 tctacaaatg tggtaaaatc gataaggatc cgtcgaccga tgcccttgag agccttcaac
     3301 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc
     3361 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac
     3421 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3481 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3541 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3601 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3661 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3721 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     3781 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3841 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3901 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3961 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4021 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4081 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4141 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4201 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4261 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4321 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4381 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4441 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4501 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4561 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4621 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4681 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4741 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4801 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4861 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4921 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4981 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5041 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5101 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5161 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5221 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5281 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc
     5341 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     5401 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5461 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     5521 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     5581 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     5641 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     5701 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5761 ttttaacaaa atattaacgc ttacaatttg ccattcgcca ttcaggctgc gcaactgttg
     5821 ggaagggcga tcggtgcggg cctcttcgct attacgccag cccaagctac catgataagt
     5881 aagtaatatt aaggtacggg aggtacttgg agcggccgc
//
