LOCUS       sequence-249886-        5042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer cloning nanotag reporter vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5042)
  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 5042)
  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_Citrine_Nanotag_24_positive_Control
FEATURES             Location/Qualifiers
     source          1..5042
                     /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        862..1007
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          1085..1217
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1200..1219
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1271..1280
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1277..1279,1280..1282,1283..1993)
                     /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(1322..1343)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1583..1602)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1930..1951
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2166..2287
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2177..2196)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2231..2250
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2535..2552)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2706..3294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2786..2805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3465..4256,4257..4325))
                     /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     4088..4107
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4326..4430)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4457..4912
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4544..4563)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4754..4775
                     /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 caggagcttt ccaccaacac ttaaccttta tgaagaacca aattattttg ttccatgact
      241 aagaaaaatc tggcgatata taattaaaac aacaaagaag agtcccattc tatacaagtg
      301 taattacgtt catgtggtat cactgtaatt gaaacacaga agatgtttca gtctgtatta
      361 aaagagaatt tctttcagtg ggtttcctga agctcattag atattccctg gcctattctc
      421 ctaagtaatt agattttaac aggattttca acagatcaga gcaatctctg tgggtgccaa
      481 taggagactc tccctctggc cttaccagca gggcagcatg gataacatcc aggctgggag
      541 ttaataggac aattactggc ctcttttaaa tgagttacat taatgcactt aagctaaatg
      601 gctctgcata tgattaaata ttaacatctg caaaatatga aatcatgata tgcaatcaag
      661 ttttgttgtt ttaatttccc aaatgtcgga ttttggtggt gtatgtaaat gtgcattgga
      721 tagtgtatgt taaactgttg gctagcccgg gctcgagatc tgcgatctaa gtaagcttgc
      781 tgcttcatcc ccgtggcccg ttgctcgcgt ttgctggcgg tgtccccgga agaaatatat
      841 ttgcatgtct ttagttctat gatgacacaa accccgccca gcgtcttgtc attggcgaat
      901 tcgaacacgc agatgcagtc ggggcggcgc ggtcccaggt ccacttcgca tattaaggtg
      961 acgcgtgtgg cctcgaacac cgagcgaccc tgcagcgacc cgcttaacag cgtcaacagc
     1021 gtgccgcaga tcccaagctt gcggccgctt aagctgcaga agttggtcgt gaggcactgg
     1081 gcaggtaagt atcaaggtta caagacaggt ttaaggagac caatagaaac tgggcttgtc
     1141 gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt
     1201 tgcctttctc tccacaggtg tccactccca ggttcaatac agctcttaag cggccgcaag
     1261 ctagcttgcc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     1321 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     1381 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     1441 cgtgccctgg cccaccctcg tgaccacctt cggctacggc ctgatgtgct tcgcccgcta
     1501 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1561 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     1621 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1681 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     1741 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     1801 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1861 gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa
     1921 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1981 cgagctgtac aagtgagcct ctagactccc tcagcggcgc gcctgtcgaa gctaaaagtg
     2041 ctcgagccca ataccatttc ggaatctcgc gtggcgttag tatgccactc gatggtaggg
     2101 gaacgcagaa tgccagatct cacgtgtcta gagtcggggc ggccggccgc ttcgagcaga
     2161 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     2221 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     2281 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     2341 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccgtcgac
     2401 cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta
     2461 tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag
     2521 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2581 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2641 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2701 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2761 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2821 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2881 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2941 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3001 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3061 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3121 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     3181 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3241 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3301 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3361 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3421 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3481 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3541 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3601 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3661 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3721 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3781 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3841 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3901 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3961 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4021 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4081 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4141 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4201 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4261 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4321 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4381 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4441 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4501 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4561 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4621 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4681 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4741 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4801 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4861 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttgccattcg
     4921 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     4981 cagcccaagc taccatgata agtaagtaat attaaggtac gggaggtact tggagcggcc
     5041 gc
//
