LOCUS       sequence-258297-        5510 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 5510)
  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 5510)
  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_Zic1_enh_373b_Citrine
FEATURES             Location/Qualifiers
     source          1..5510
                     /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        1280..1425
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1503..1635
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1618..1637
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1689..1698
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1695..1697,1698..1700,1701..2411)
                     /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(1740..1761)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2001..2020)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2348..2369
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2585..2706
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2596..2615)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2650..2669
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2954..2971)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3125..3713)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3205..3224)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3884..4675,4676..4744))
                     /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     4507..4526
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4745..4849)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4876..5331
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4963..4982)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5173..5194
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5462..5510
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 aatcgatagt actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
       61 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctctatcga taggtaccga
      121 gctcgagttg ccaggaatgt ggtgtgggta gtgaaaaata acattaaaaa aaatgctgta
      181 acatgttttt atttctattg gttcaaaaca gcatttgctt cctgaaatta aatggaatcc
      241 aattgaggtt ctgtcctttc agagacctag tcttgtacaa ggtaatgcat tagtgactac
      301 tctattgata ttgtaatttt tctagcatac gtagaaattg tcatattgca agttggtggg
      361 gcccttgata tttcacaaat tggagatcat tacaagtgtt ttccatgcca gagctatact
      421 aattgtgacc atgttgatga catccaccat tctacttaaa tagaaaatcg ctgcttacta
      481 aagaagttgg acttagctta agtccttggt gactgatttg attagtccat ttgagtgaga
      541 atagggcaac acaggggcta tgttaatgta acttaaacaa taggccatca ctaatcagag
      601 acctcaaggt tgaagtgagg ttaattgtta tgcgggtggt atttgaaaag aagtgttgac
      661 tggtgtaatg cactatggga ctgtagtatt tgaacatgca gccctaaggc cagaggtttt
      721 gtttagtttc ccaatgaaat ttaaagctac attatgtaca tctctaaaca acagcctata
      781 cgctactgcc ttttgataca tcacaagagc aactactaat tgataacaga aatgagcccc
      841 caggcacaac attcagatcc aaatctggta ctggatttaa atccctttct aaagttggtg
      901 aggtgggaat tgattttgat taagcccttc atggaatgaa cacgcaagat tcaaatgact
      961 cctttctgcc cctgaaagtt taagaaattc caaattcaca gtttcagttt agtcttatag
     1021 tatgcattta gttgaaacat atgagctgac aacatcttaa tgcaaaaaca tcttggcata
     1081 attacagagg gctgtttatt gatatcattg ttcggacatg gatttatgtc tcaacgtgag
     1141 tgccaggagt ttctgttggc tagcccgggc tcgagatctg cgatctaagt aagcttgctg
     1201 cttcatcccc gtggcccgtt gctcgcgttt gctggcggtg tccccggaag aaatatattt
     1261 gcatgtcttt agttctatga tgacacaaac cccgcccagc gtcttgtcat tggcgaattc
     1321 gaacacgcag atgcagtcgg ggcggcgcgg tcccaggtcc acttcgcata ttaaggtgac
     1381 gcgtgtggcc tcgaacaccg agcgaccctg cagcgacccg cttaacagcg tcaacagcgt
     1441 gccgcagatc ccaagcttgc ggccgcttaa gctgcagaag ttggtcgtga ggcactgggc
     1501 aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga
     1561 gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg
     1621 cctttctctc cacaggtgtc cactcccagg ttcaatacag ctcttaagcg gccgcaagct
     1681 agcttgccgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1741 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1801 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1861 tgccctggcc caccctcgtg accaccttcg gctacggcct gatgtgcttc gcccgctacc
     1921 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1981 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2041 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2101 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     2161 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     2221 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2281 tgcccgacaa ccactacctg agctaccagt ccgccctgag caaagacccc aacgagaagc
     2341 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     2401 agctgtacaa gtgagcctct agactccctc agcggcgcgc cgttgcgttc caaacgtcgt
     2461 ggagctccct gtcaagacac gaaccgctaa cgccacctta gtccgtgaag catgatacgc
     2521 gacatacccc aggagatctc acgtgtctag agtcggggcg gccggccgct tcgagcagac
     2581 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     2641 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     2701 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     2761 gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tccgtcgacc
     2821 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     2881 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     2941 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3001 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3061 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3121 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3181 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3241 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3301 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3361 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3421 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3481 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3541 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3601 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3661 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3721 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3781 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3841 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3901 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3961 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4021 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4081 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4141 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4201 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4261 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4321 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4381 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4441 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4501 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4561 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4621 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4681 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4741 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4801 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4861 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     4921 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4981 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5041 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     5101 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5161 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     5221 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     5281 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tgccattcgc
     5341 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     5401 agcccaagct accatgataa gtaagtaata ttaaggtacg ggaggtactt ggagcggccg
     5461 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg
//
