LOCUS       sequence-257594-        4527 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 4527)
  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 4527)
  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_Cerulean_Nanotag_62_neg_Control
FEATURES             Location/Qualifiers
     source          1..4527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..169
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          247..379
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     362..381
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      433..442
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(439..441,442..444,445..1158)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(484..505)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(745..764)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1092..1113
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1330..1451
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1341..1360)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1395..1414
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1699..1716)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1870..2458)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1950..1969)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2629..3420,3421..3489))
                     /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     3252..3271
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3490..3594)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3621..4076
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3708..3727)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3918..3939
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4207..4255
                     /note="synthetic polyadenylation signal"
     misc_feature    4269..4360
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4309..4328
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 atttgcatgt ctttagttct atgatgacac aaaccccgcc cagcgtcttg tcattggcga
       61 attcgaacac gcagatgcag tcggggcggc gcggtcccag gtccacttcg catattaagg
      121 tgacgcgtgt ggcctcgaac accgagcgac cctgcagcga cccgcttaac agcgtcaaca
      181 gcgtgccgca gatcccaagc ttgcggccgc ttaagctgca gaagttggtc gtgaggcact
      241 gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg
      301 tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac
      361 tttgcctttc tctccacagg tgtccactcc caggttcaat acagctctta agcggccgca
      421 agctagcttg ccgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      481 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      541 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      601 cccgtgccct ggcccaccct cgtgaccacc ctgacctggg gcgtgcagtg cttcgcccgc
      661 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      721 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
      781 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
      841 ggcaacatcc tggggcacaa gctggagtac aacgccatca gcgacaacgt ctatatcacc
      901 gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac
      961 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1021 ctgctgcccg acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag
     1081 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1141 gacgagctgt acaagtgagc ctctagactc cctcagcggc gcgccacttt tcggttctgg
     1201 tggacgtaat caatcagcga acgccgtctt ctgccgtctt ctggtaacgc acgaactctg
     1261 ccgagcaacc tacagaccag atctcacgtg tctagagtcg gggcggccgg ccgcttcgag
     1321 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     1381 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     1441 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     1501 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatccgt
     1561 cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg
     1621 actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg
     1681 gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     1741 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     1801 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1861 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1921 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1981 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2041 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2101 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2161 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2221 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2281 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2341 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2401 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2461 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2521 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2581 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2641 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     2701 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     2761 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     2821 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     2881 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     2941 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3001 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3061 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3121 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3181 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3241 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     3301 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     3361 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     3421 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     3481 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     3541 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     3601 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3661 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3721 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3781 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3841 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3901 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3961 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4021 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttgcca
     4081 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     4141 acgccagccc aagctaccat gataagtaag taatattaag gtacgggagg tacttggagc
     4201 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg
     4261 atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag
     4321 gctgtcccca gtgcaagtgc aggtgccaga acatttctct atcgataggt accgagctcg
     4381 agttccatga gctggatcga tgatatccga tcgatcgtag cacaggaccg ctagcccggg
     4441 ctcgagatct gcgatctaag taagcttgct gcttcatccc cgtggcccgt tgctcgcgtt
     4501 tgctggcggt gtccccggaa gaaatat
//
