LOCUS       sequence-249955-        5509 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 5509)
  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 5509)
  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..5509
                     /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        1328..1473
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1551..1683
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1666..1685
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1737..1746
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1743..1745,1746..1748,1749..2459)
                     /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(1788..1809)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2049..2068)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2396..2417
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2633..2754
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2644..2663)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2698..2717
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3002..3019)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3173..3761)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3253..3272)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3932..4723,4724..4792))
                     /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     4555..4574
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4793..4897)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4924..5379
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5011..5030)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5221..5242
                     /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 caggaatgtg gtgtgggtag tgaaaaataa cattaaaaaa atgctgtaac atgtttttat
      241 ttctattggt tcaaaacagc atttgcttcc tgaaattaaa tggaatccaa ttgaggttct
      301 gtcctttcag agacctagtc ttgtacaagg taatgcatta gtgactactc tattgatatt
      361 gtaatttttc tagcatacgt agaaattgtc atattgcaag ttggtggggc ccttgatatt
      421 tcacaaattg gagatcatta caagtgtttt ccatgccaga gctatactaa ttgtgaccat
      481 gttgatgaca tccaccattc tacttaaata gaaaatcgct gcttactaaa gaagttggac
      541 ttagcttaag tccttggtga ctgatttgat tagtccattt gagtgagaat agggcaacac
      601 aggggctatg ttaatgtaac ttaaacaata ggccatcact aatcagagac ctcaaggttg
      661 aagtgaggtt aattgttatg cgggtggtat ttgaaaagaa gtgttgactg gtgtaatgca
      721 ctatgggact gtagtatttg aacatgcagc cctaaggcca gaggttttgt ttagtttccc
      781 aatgaaattt aaagctacat tatgtacatc tctaaacaac agcctatacg ctactgcctt
      841 ttgatacatc acaagagcaa ctactaattg ataacagaaa tgagccccca ggcacaacat
      901 tcagatccaa atctggtact ggatttaaat ccctttctaa agttggtgag gtgggaattg
      961 attttgatta agcccttcat ggaatgaaca cgcaagattc aaatgactcc tttctgcccc
     1021 tgaaagttta agaaattcca aattcacagt ttcagtttag tcttatagta tgcatttagt
     1081 tgaaacatat gagctgacaa catcttaatg caaaaacatt ttggcataat tacagagggc
     1141 tgtttattga tatcattgtt cggacatgga tttatgtctt aacgtgagtg ccaggagttt
     1201 ctgttggcta gcccgggctc gagatctgcg atctaagtaa gcttgctgct tcatccccgt
     1261 ggcccgttgc tcgcgtttgc tggcggtgtc cccggaagaa atatatttgc atgtctttag
     1321 ttctatgatg acacaaaccc cgcccagcgt cttgtcattg gcgaattcga acacgcagat
     1381 gcagtcgggg cggcgcggtc ccaggtccac ttcgcatatt aaggtgacgc gtgtggcctc
     1441 gaacaccgag cgaccctgca gcgacccgct taacagcgtc aacagcgtgc cgcagatccc
     1501 aagcttgcgg ccgcttaagc tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca
     1561 aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac
     1621 tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca
     1681 caggtgtcca ctcccaggtt caatacagct cttaagcggc cgcaagctag cttgccgcca
     1741 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1801 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     1861 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     1921 ccctcgtgac caccttcggc tacggcctga tgtgcttcgc ccgctacccc gaccacatga
     1981 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     2041 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     2101 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     2161 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     2221 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     2281 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     2341 actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     2401 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     2461 gagcctctag actccctcag cggcgcgccg ttgcgttcca aacgtcgtgg agctccctgt
     2521 caagacacga accgctaacg ccaccttagt ccgtgaagca tgatacgcga cataccccag
     2581 gagatctcac gtgtctagag tcggggcggc cggccgcttc gagcagacat gataagatac
     2641 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2701 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2761 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     2821 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtcgaccga tgcccttgag
     2881 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     2941 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt
     3001 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3061 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3121 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3181 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3241 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3301 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3361 tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3421 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3481 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3541 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3601 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3661 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3721 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3781 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3841 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3901 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3961 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4021 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4081 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4141 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4201 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4261 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4321 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4381 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4441 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4501 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4561 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4621 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4681 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4741 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4801 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4861 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4921 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4981 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     5041 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     5101 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     5161 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     5221 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     5281 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     5341 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg ccattcgcca ttcaggctgc
     5401 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag cccaagctac
     5461 catgataagt aagtaatatt aaggtacggg aggtacttgg agcggccgc
//
