LOCUS       sequence-265027-        6282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses YFP-NOP58 fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6282)
  AUTHORS   Newhart A, Powers SL, Shastrula PK, Sierra I, Joo LM, Hayden JE, 
            Cohen AR, Janicki SM
  TITLE     RNase P protein subunit Rpp29 represses histone H3.3 nucleosome 
            deposition.
  JOURNAL   Mol Biol Cell. 2016 Apr 1;27(7):1154-69. doi: 
            10.1091/mbc.E15-02-0099. Epub 2016 Feb 3.
  PUBMED    26842893
REFERENCE   2  (bases 1 to 6282)
  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: 
            "10.1091/mbc.E15-02-0099"; journalName: "Mol Biol Cell"; date: 
            "2016-04-1- 1"; volume: "27"; issue: "7"; pages: "1154-69"
COMMENT     Sequence Label: YFP-NOP58-C1
FEATURES             Location/Qualifiers
     source          1..6282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      243..252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(249..251,252..254,255..965)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(294..315)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(555..574)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     902..923
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2706..2827
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2743..2762)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2797..2816
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2834..3289)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2971..2992)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3183..3202
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3316..3420
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3422..3779
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3422..3613
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3445..3465)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3630..3765
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3814..4608
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3814..4608
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3868..3887)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4478..4497
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4796..4815)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4840..4887
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5216..5804
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5705..5724
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5979..6282
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      301 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      361 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      421 ggcccaccct cgtgaccacc ttcggctacg gcctgaagtg cttcgcccgc taccccgacc
      481 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      541 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      601 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      661 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
      721 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      781 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      841 acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
      901 acatggtcct gctggagttc gtgaccgccg ccgggatcac tcacggcatg gacgagctgt
      961 acaagtccgg actcagatct cgagctatgt tggtgctgtt tgaaacgtct gtgggttacg
     1021 ccatctttaa ggttctaaat gagaagaaac ttcaagaggt tgatagttta tggaaagaat
     1081 ttgaaactcc agagaaagca aacaaaatag taaagctaaa acattttgag aaatttcagg
     1141 atacagcaga agcattagca gcattcacag ctctgatgga gggcaaaatc aataagcagc
     1201 tgaaaaaagt tctgaagaaa atagtaaaag aagcccatga accgctggca gtagctgatg
     1261 ctaaactagg aggggtcata aaggaaaagc tgaatctcag ttgtatccat agtcctgttg
     1321 ttaatgaact tatgagagga attcgttcac aaatggatgg attaatccct ggggtagaac
     1381 cacgtgaaat ggcagctatg tgtcttggat tggctcacag cctgtctcga tatagattga
     1441 agtttagcgc tgataaagta gacacaatga ttgttcaggc aatttccttg ttagatgact
     1501 tggataaaga actaaacaac tacattatgc gatgtagaga atggtatggc tggcatttcc
     1561 ctgaattagg aaaaattatt tcagataatt taacatactg caagtgttta cagaaagttg
     1621 gcgataggaa gaactatgcc tctgccaagc tttctgagtt gctgccagaa gaagttgaag
     1681 cagaagtgaa agcagctgca gagatatcaa tgggaacaga ggtttcagaa gaagatattt
     1741 gcaatattct gcatctttgc acccaggtga ttgaaatctc tgaatatcga acccagctct
     1801 atgaatatct acaaaatcga atgatggcca ttgcacccaa tgttacagtc atggttgggg
     1861 aattagttgg agcacggctt attgctcatg caggttctct tttaaatttg gccaagcatg
     1921 cagcttctac cgttcagatt cttggagctg aaaaggcact tttcagagcc ctcaaatcta
     1981 gacgggatac ccctaagtat ggtctcattt atcatgcttc actcgtgggc cagacaagtc
     2041 ccaaacacaa aggaaagatt tctcgaatgc tggcagccaa aaccgttttg gctatccgtt
     2101 atgatgcttt tggtgaggat tcaagttctg caatgggagt tgagaacaga gccaaattag
     2161 aggccaggtt gagaactttg gaagacagag ggataagaaa aataagtgga acaggaaaag
     2221 cattagcaaa aacagaaaaa tatgaacaca aaagtgaagt gaagacttac gatccttctg
     2281 gtgactccac acttccaacc tgctctaaaa aacgcaaaat agaacaggta gataaagagg
     2341 atgaaattac tgaaaagaaa gccaaaaaag ccaagattaa agttaaagtt gaagaagagg
     2401 aagaagaaaa agtggcagaa gaagaagaaa catctgtgaa gaagaagaag aaaaggggta
     2461 aaaagaaaca cattaaggaa gaaccacttt ctgaggaaga accatgtacc agcacagcaa
     2521 ttgctagtcc agagaaaaag aagaaaaaga aaaaaaagag agagaacgag gattaaggat
     2581 ccaccggatc tagataactg atcataatca gccataccac atttgtagag gttttacttg
     2641 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     2701 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     2761 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     2821 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     2881 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     2941 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     3001 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     3061 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     3121 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     3181 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     3241 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     3301 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3361 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3421 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3481 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3541 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3601 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3661 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     3721 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     3781 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3841 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3901 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3961 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     4021 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4081 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4141 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4201 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4261 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4321 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     4381 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4441 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4501 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4561 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4621 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4681 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4741 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     4801 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     4861 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     4921 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     4981 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     5041 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     5101 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5161 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5221 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5281 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5341 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5401 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5461 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5521 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5581 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5641 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5701 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5761 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5821 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5881 cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata gtaatcaatt
     5941 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     6001 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     6061 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     6121 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     6181 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     6241 acttggcagt acatctacgt attagtcatc gctattacca tg
//
