LOCUS       sequence-257651-        4523 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 4523)
  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 4523)
  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_18_neg_Control
FEATURES             Location/Qualifiers
     source          1..4523
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..146
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          224..356
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     339..358
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      410..419
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(416..418,419..421,422..1132)
                     /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(461..482)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(722..741)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1069..1090
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1307..1428
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1318..1337)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1372..1391
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1676..1693)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1847..2435)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1927..1946)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2606..3397,3398..3466))
                     /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     3229..3248
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3467..3571)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3598..4053
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3685..3704)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3895..3916
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4184..4232
                     /note="synthetic polyadenylation signal"
     misc_feature    4246..4337
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4286..4305
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 atgacacaaa ccccgcccag cgtcttgtca ttggcgaatt cgaacacgca gatgcagtcg
       61 gggcggcgcg gtcccaggtc cacttcgcat attaaggtga cgcgtgtggc ctcgaacacc
      121 gagcgaccct gcagcgaccc gcttaacagc gtcaacagcg tgccgcagat cccaagcttg
      181 cggccgctta agctgcagaa gttggtcgtg aggcactggg caggtaagta tcaaggttac
      241 aagacaggtt taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg
      301 tttctgatag gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt
      361 ccactcccag gttcaataca gctcttaagc ggccgcaagc tagcttgccg ccaccatggt
      421 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
      481 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
      541 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
      601 gaccaccttc ggctacggcc tgatgtgctt cgcccgctac cccgaccaca tgaagcagca
      661 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
      721 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
      781 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
      841 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
      901 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
      961 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1021 gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1081 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtgagcctc
     1141 tagactccct cagcggcgcg ccttgcagta attcacgagg ccaaaggcga accatttcca
     1201 ccagcggtac tccggctcat tcttaccgcg cagatcgttg tggacagatc gcgaaagatc
     1261 tcacgtgtct agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat
     1321 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     1381 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     1441 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     1501 aacctctaca aatgtggtaa aatcgataag gatccgtcga ccgatgccct tgagagcctt
     1561 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     1621 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc
     1681 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1741 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1801 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1861 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1921 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1981 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2041 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2101 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2161 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2221 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2281 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2341 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2401 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2461 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2521 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2581 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2641 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     2701 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     2761 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     2821 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     2881 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     2941 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3001 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3061 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3121 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3181 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     3241 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     3301 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     3361 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     3421 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     3481 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3541 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     3601 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3661 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3721 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3781 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3841 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3901 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3961 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4021 cgaattttaa caaaatatta acgcttacaa tttgccattc gccattcagg ctgcgcaact
     4081 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagcccaag ctaccatgat
     4141 aagtaagtaa tattaaggta cgggaggtac ttggagcggc cgcaataaaa tatctttatt
     4201 ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gtactaacat acgctctcca
     4261 tcaaaacaaa acgaaacaaa acaaactagc aaaataggct gtccccagtg caagtgcagg
     4321 tgccagaaca tttctctatc gataggtacc gagctcgagt tgccaggtgc tacgatcgat
     4381 cggatatcat cgatccagct tggctagccc gggctcgaga tctgcgatct aagtaagctt
     4441 gctgcttcat ccccgtggcc cgttgctcgc gtttgctggc ggtgtccccg gaagaaatat
     4501 atttgcatgt ctttagttct atg
//
