LOCUS       sequence-259316-        6908 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 6908)
  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 6908)
  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_Ets1_enh_117_Citrine
FEATURES             Location/Qualifiers
     source          1..6908
                     /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        2677..2822
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          2900..3032
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3015..3034
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      3086..3095
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3092..3094,3095..3097,3098..3808)
                     /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(3137..3158)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3398..3417)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3745..3766
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3983..4104
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3994..4013)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4048..4067
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4352..4369)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4523..5111)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4603..4622)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5282..6073,6074..6142))
                     /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     5905..5924
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6143..6247)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6274..6729
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6361..6380)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6571..6592
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6860..6908
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
       61 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
      121 gctcgagttg ccaggtgctc gctgttaatg cctctgctcc ctcaggacta ggccaagtga
      181 tggggttaat ttgggagggg gctggaggtg cagcttcagg agatggcaat taacaggaga
      241 cagcgctcgg ggaactcact gcaaatgaaa aaacaacagg tcctcgctcc tgtggtgtta
      301 aggagtggct taagggggtt gtccggagcc cagaaatctg ggagtaattt gggacaaaac
      361 aatgtcctct gcaccacttt gatgctgcag aaacatgaga acatgcagag atagcgtcgc
      421 atctcagagc ctccccaccc caaggccgtg gagatttaga tttagatcct gatcagggcc
      481 tgcagtgtgc tgcaatcaat acagccaagg ctgaaggcga tttaattgtg gcaagtaatg
      541 acccaacacc ctggggagat gcttgtggat ggaagtcgag gaatttgctc agatctgtgc
      601 agaggtgggc aatgggatga gcccaagctt ggagttttta cccttttttg gaagaaaaat
      661 gtctctccac ctctgttttt tccccttaac tcaaaaatca tcatctattc tccgtgagag
      721 aatgggcagc tgaatatcac gaatggcata attccaggaa aagctgctcc cttttatact
      781 gcagagagga atgtggaaaa gcaattcaaa aagcccaaga gaaataatta tcaattttca
      841 gccacagcag cctggagcca atggggcaca catggcagag ctcagcttag ggctgccatg
      901 gctgaacgct tgtgccaata gagcaagtgt aggtgggtgc caatacagct gaagtacaca
      961 tcataaatag gtgtaagcgt cctgacggtt gcagaagtgc atgaacactt ggtgttacaa
     1021 ggttgtgaac ttctgggagt ctcatagtga aataccaaat aaccactggt gccttttaat
     1081 tttggctctt ctctgactat ttactcttgc tgggtttttt tttttagcta taaccttctg
     1141 gagggtgtgg gcgatgctgt ttttaaggtg accgcggagg atggctccag catcactcaa
     1201 aagaactttc tgggccctct agtggtgtgt gctgtgctga ggacctgatc tccctatcct
     1261 tttgttccac caaaataaaa actagtcatt attttcccca ttatttttca caaaatgagg
     1321 aaaatcccaa atctccctgg tcttattttt tatttttatt cttcttagaa cgcacctatg
     1381 gaaggagcag tgcatccgtt gggttaaaac cctccctctg cctcttcaga accaacgaac
     1441 gtatgcagac attcatttca aaactgaaaa cccctctcga aaagacaaag gtgggctgat
     1501 atgttttgag gacatcaggg caatggggga ctcatgcctt tctatcattt gtttctcctg
     1561 ttatggcttg tgaagaaaat aatgtcacag gggtcttatc agggagctgg ggaaggacag
     1621 gggatttggg aagagcttac gtatttcttt ccaagctgtg tctctaaaat aagttgatgt
     1681 ttatttggtt gcgagggaaa aacgtttaat ttgatatcct gtccgtgctg gaaacagcaa
     1741 acattttaac aaaggggagg atgggaaaag gtggctggaa aagcagcgtc tgtaggtgtt
     1801 tgggctttgt ttgttgagta acaccattgc tgacagcatc atccccagcc accctgcctg
     1861 cgtacaccag cactcatccc cagggcgcac acagctttgt gtcctgccta ccccacacaa
     1921 ctatttgtgc tctgccaggt aggggtgtga gttttgactg aaatggctct ttctggtggt
     1981 ttagcagatg taggcagagg cagcatgagc tgcaaaggcc ttggggtggg ttctgccctt
     2041 tactacctgg gcatccagcc tttcttttgg atttttgaca ccttcatagg cttgctcaag
     2101 aggtaaatta agggtgttaa gatggatatc cagggatctt gggatagatt taggggaaac
     2161 ctccccttca tattgagctc ccagcttaag caatgctctt ccacatcttt ccctgttggc
     2221 ctgcatggga tggtgcaggg tgcagccatg ttgaggtgtc cccttgggtt caagactgaa
     2281 tttttccaag tgtggacgct gcagcagact gctctggagg gctcagcata agggatccca
     2341 aaaccttcac cttgccaacc cagcctgctg gttttagtgg tttgcaaatg caaaccagct
     2401 tgactgtgct gctttggcag ccgtaggagc ttcacatcac catcctgatt gcatttggat
     2461 ttgggcactg tagtcctgga aaacacttgg agcagccaac atcaaatgta acaaagcaag
     2521 acctggatgt gtgaggatgg agtgatgccc tgttggctag cccgggctcg agatctgcga
     2581 tctaagtaag cttgctgctt catccccgtg gcccgttgct cgcgtttgct ggcggtgtcc
     2641 ccggaagaaa tatatttgca tgtctttagt tctatgatga cacaaacccc gcccagcgtc
     2701 ttgtcattgg cgaattcgaa cacgcagatg cagtcggggc ggcgcggtcc caggtccact
     2761 tcgcatatta aggtgacgcg tgtggcctcg aacaccgagc gaccctgcag cgacccgctt
     2821 aacagcgtca acagcgtgcc gcagatccca agcttgcggc cgcttaagct gcagaagttg
     2881 gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag gagaccaata
     2941 gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct
     3001 tactgacatc cactttgcct ttctctccac aggtgtccac tcccaggttc aatacagctc
     3061 ttaagcggcc gcaagctagc ttgccgccac catggtgagc aagggcgagg agctgttcac
     3121 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     3181 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     3241 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgat
     3301 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     3361 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     3421 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     3481 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     3541 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     3601 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     3661 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa
     3721 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     3781 cactctcggc atggacgagc tgtacaagtg agcctctaga ctccctcagc ggcgcgccac
     3841 caagtcagtg attgcgcgag cgatgttgga gtgctcctac tgcaggtacg aaggtcttat
     3901 cggatcgccc gaacaccaca cagtgacctt gagatctcac gtgtctagag tcggggcggc
     3961 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     4021 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     4081 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     4141 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     4201 gataaggatc cgtcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg
     4261 ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt
     4321 aggacaggtg ccggcagcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4381 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4441 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4501 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4561 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4621 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4681 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4741 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4801 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4861 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4921 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4981 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5041 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5101 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5161 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5221 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5281 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5341 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5401 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5461 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5521 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5581 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5641 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5701 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5761 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5821 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5881 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5941 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6001 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6061 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6121 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6181 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6241 ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg
     6301 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     6361 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     6421 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     6481 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     6541 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     6601 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     6661 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     6721 ttacaatttg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     6781 cctcttcgct attacgccag cccaagctac catgataagt aagtaatatt aaggtacggg
     6841 aggtacttgg agcggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt
     6901 tttgtgtg
//
