LOCUS       sequence-257558-        5047 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 5047)
  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 5047)
  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_92_positive_Control
FEATURES             Location/Qualifiers
     source          1..5047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      24..612
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     513..532
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     766..783
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    1031..1152
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1068..1087)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1122..1141
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(1327..2040,2041..2043,2044..2046))
                     /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(1372..1393)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1721..1740
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1980..2001
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2043..2052
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2104..2123)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          2106..2238
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(2316..2461)
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     misc_feature    3169..3260
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(3201..3220)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    3274..3322
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(3453..3908)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3590..3611)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3802..3821
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3935..4039
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4040..4108,4109..4900)
                     /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     complement(4258..4277)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
       61 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
      121 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
      181 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
      241 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
      301 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
      361 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
      421 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
      481 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
      541 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
      601 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
      661 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
      721 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
      781 gaggaagcgg aagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt
      841 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
      901 ggctctcaag ggcatcggtc gacggatcct tatcgatttt accacatttg tagaggtttt
      961 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     1021 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     1081 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     1141 caatgtatct tatcatgtct gctcgaagcg gccggccgcc ccgactctag acacgtgaga
     1201 tctaccatta gactggcttg taccggaaag cggactgtac tttgcgtgaa gtaaacagct
     1261 gccgataatg accgtggtac cacccggtac agacagcaag gcgcgccgct gagggagtct
     1321 agaggctcac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc
     1381 cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagcttgg actgggtgct
     1441 caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta
     1501 gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttggcctt
     1561 gatgccgttc ttctgcttgt cggcggtgat atagacgttg tcgctgatgg cgttgtactc
     1621 cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg
     1681 gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc
     1741 cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc
     1801 gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc acgccccagg tcagggtggt
     1861 cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag
     1921 cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac
     1981 gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct
     2041 caccatggtg gcggcaagct agcttgcggc cgcttaagag ctgtattgaa cctgggagtg
     2101 gacacctgtg gagagaaagg caaagtggat gtcagtaaga ccaataggtg cctatcagaa
     2161 acgcaagagt cttctctgtc tcgacaagcc cagtttctat tggtctcctt aaacctgtct
     2221 tgtaaccttg atacttacct gcccagtgcc tcacgaccaa cttctgcagc ttaagcggcc
     2281 gcaagcttgg gatctgcggc acgctgttga cgctgttaag cgggtcgctg cagggtcgct
     2341 cggtgttcga ggccacacgc gtcaccttaa tatgcgaagt ggacctggga ccgcgccgcc
     2401 ccgactgcat ctgcgtgttc gaattcgcca atgacaagac gctgggcggg gtttgtgtca
     2461 tcatagaact aaagacatgc aaatatattt cttccgggga caccgccagc aaacgcgagc
     2521 aacgggccac ggggatgaag cagcaagctt acttagatcg cagatctcga gcccgggcta
     2581 gcggtccttt taacatacac tatccaatgc acatttacat acaccaccaa aatccggcat
     2641 ttgggaaatt aaaacaacaa aacttgattg catatcatga tttcatattt tgcagatgtt
     2701 aatatttaat catatgcaga gccatttagc ttaagtgcat taatgtaact catttaaaag
     2761 aggccagtaa tcgtcctatt gactcccagc ctggatgtta tccatgctgc cctgctggta
     2821 aggccagagg gagagtctcc tattggcacc cacagagatt gctctgatct gttgaaaatc
     2881 ctgttaaaat ctaattactt aggagaatag gccagggaat atctaatgag cttcaggaaa
     2941 cccactgaaa gaaattctct tttaatacag actgaaacat cttctgtgtt tcaattacag
     3001 tgataccaca tgaacgtaat tacacttgta tagaatggga ctcttctttg ttgttttaat
     3061 tatatatcgc cagatttttc ttagtcatgg aacaaaataa tttggttctt cataaaggtt
     3121 aagtgttggt ggaaagctcc atggaactcg agctcggtac ctatcgatag agaaatgttc
     3181 tggcacctgc acttgcactg gggacagcct attttgctag tttgttttgt ttcgttttgt
     3241 tttgatggag agcgtatgtt agtactatcg attcacacaa aaaaccaaca cacagatgta
     3301 atgaaaataa agatatttta ttgcggccgc tccaagtacc tcccgtacct taatattact
     3361 tacttatcat ggtagcttgg gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     3421 ccaacagttg cgcagcctga atggcgaatg gcaaattgta agcgttaata ttttgttaaa
     3481 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3541 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3601 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3661 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     3721 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3781 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3841 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3901 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3961 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4021 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4081 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4141 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4201 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4261 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4321 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4381 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4441 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4501 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4561 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4621 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4681 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4741 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4801 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4861 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4921 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4981 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5041 agacccc
//
