LOCUS       sequence-134195-        7897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression clone for human NOP58 with C-terminal 3xFLAG-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7897)
  AUTHORS   Falaleeva M, Pages A, Matuszek Z, Hidmi S, Agranat-Tamir L, Korotkov
            K, Nevo Y, Eyras E, Sperling R, Stamm S
  TITLE     Dual function of C/D box small nucleolar RNAs in rRNA modification 
            and alternative pre-mRNA splicing.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Mar 22;113(12):E1625-34. doi: 
            10.1073/pnas.1519292113. Epub 2016 Mar 8.
  PUBMED    26957605
REFERENCE   2  (bases 1 to 7897)
  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: 
            "10.1073/pnas.1519292113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-03-22- 22"; volume: "113"; issue: "12"; pages: 
            "E1625-34"
COMMENT     Sequence Label: pCMV14_NOP58
FEATURES             Location/Qualifiers
     source          1..7897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2580..2645
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2656..3278
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2855..2874)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3302..3322)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3320..3617
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3671..4465
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3725..3744)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4335..4354
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5117..5251
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5154..5173)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5208..5227
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5256..5275)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5257..5275)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5289..5305)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5289..5305)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5302..5324)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5313..5329
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5337..5343,5344..5361,5362..5367))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5382..5403
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5520..5537)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5691..6279)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5771..5790)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6450..7241,7242..7310))
                     /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     7073..7092
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7311..7415)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7442..7870
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7529..7548)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7739..7760
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaagcttg cggccgcgaa ttcatcgata gatctgatat
      961 cggtaccagt cgacatgttg gtgctgtttg aaacgtctgt gggttacgcc atctttaagg
     1021 ttctaaatga gaagaaactt caagaggttg atagtttatg gaaagaattt gaaactccag
     1081 agaaagcaaa caaaatagta aagctaaaac attttgagaa atttcaggat acagcagaag
     1141 cattagcagc attcacagct ctgatggagg gcaaaatcaa taagcagctg aaaaaagttc
     1201 tgaagaaaat agtaaaagaa gcccatgaac cgctggcagt agctgatgct aaactaggag
     1261 gggtcataaa ggaaaagctg aatctcagtt gtatccatag tcctgttgtt aatgaactta
     1321 tgagaggaat tcgttcacaa atggatggat taatccctgg ggtagaacca cgtgaaatgg
     1381 cagctatgtg tcttggattg gctcacagcc tgtctcgata tagattgaag tttagcgctg
     1441 ataaagtaga cacaatgatt gttcaggcaa tttccttgtt agatgacttg gataaagaac
     1501 taaacaacta cattatgcga tgtagagaat ggtatggctg gcatttccct gaattaggaa
     1561 aaattatttc agataattta acatactgca agtgtttaca gaaagttggc gataggaaga
     1621 actatgcctc tgccaagctt tctgagttgc tgccagaaga agttgaagca gaagtgaaag
     1681 cagctgcaga gatatcaatg ggaacagagg tttcagaaga agatatttgc aatattctgc
     1741 atctttgcac ccaggtgatt gaaatctctg aatatcgaac ccagctctat gaatatctac
     1801 aaaatcgaat gatggccatt gcacccaatg ttacagtcat ggttggggaa ttagttggag
     1861 cacggcttat tgctcatgca ggttctcttt taaatttggc caagcatgca gcttctaccg
     1921 ttcagattct tggagctgaa aaggcacttt tcagagccct caaatctaga cgggataccc
     1981 ctaagtatgg tctcatttat catgcttcac tcgtgggcca gacaagtccc aaacacaaag
     2041 gaaagatttc tcgaatgctg gcagccaaaa ccgttttggc tatccgttat gatgcttttg
     2101 gtgaggattc aagttctgca atgggagttg agaacagagc caaattagag gccaggttga
     2161 gaactttgga agacagaggg ataagaaaaa taagtggaac aggaaaagca ttagcaaaaa
     2221 cagaaaaata tgaacacaaa agtgaagtga agacttacga tccttctggt gactccacac
     2281 ttccaacctg ttctaaaaaa cgcaaaatag aacaggtaga taaagaggat gaaattactg
     2341 aaaagaaagc caaaaaagcc aagattaaag ttaaagttga agaagaggaa gaagaaaaag
     2401 tggcagaaga agaagaaaca tctgtgaaga agaagaagaa aaggggtaaa aagaaacaca
     2461 ttaaggaaga accactttct gaggaagaac catgtaccag cacagcaatt gctagtccag
     2521 agaaaaagaa gaaaaagaaa aaaaagagag agaacgagga ttctagagga tcccgggctg
     2581 actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg
     2641 acaagtagtg atcccgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     2701 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     2761 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     2821 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     2881 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     2941 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     3001 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     3061 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     3121 ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca gacacagcat
     3181 aggctacctg gccatgccca accggtggga catttgagtt gcttgcttgg cactgtcctc
     3241 tcatgcgttg ggtccactca gtagatgcct gttgaattgg gtacgcggcc agcttggctg
     3301 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3361 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3421 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3481 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3541 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3601 tgaggaggct tttttggagg aattgatcag cttgggatct gatcaagaga caggatgagg
     3661 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3721 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3781 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3841 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3901 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3961 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4021 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4081 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4141 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4201 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4261 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4321 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4381 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4441 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4501 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4561 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4621 gagttcttcg cccaccccgg gctcgatccc ctcgcgagtt ggttcagctg ctgcctgagg
     4681 ctggacgacc tcgcggagtt ctaccggcag tgcaaatccg tcggcatcca ggaaaccagc
     4741 agcggctatc cgcgcatcca tgcccccgaa ctgcaggagt ggggaggcac gatggccgct
     4801 ttggtcgacc cggacgggac gctcctgcgc ctgatacaga acgaattgct tgcaggcatc
     4861 tcatgagtgt gtcttcccgt tttccgcctg aggtcactgc gtggatggag cgctggcgcc
     4921 tgctgcgcga cggcgagctg ctcaccaccc actcgccaag ctggaaccgt aaaaaggccg
     4981 cgttgctggc gtttttccat aggctccgcc gatcataatc agccatacca catttgtaga
     5041 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     5101 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5161 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5221 actcatcaat gtatcttatc atgtctggat caattcccta tagtgagtcg tattaaattc
     5281 gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5341 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5401 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5461 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5521 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5581 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5641 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5701 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5761 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5821 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5881 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5941 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6001 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6061 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6121 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6181 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6241 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6301 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6361 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6421 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6481 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6541 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6601 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6661 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6721 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6781 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6841 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6901 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6961 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7021 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7081 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7141 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7201 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7261 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7321 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7381 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7441 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     7501 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     7561 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     7621 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     7681 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     7741 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     7801 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     7861 aacgcgaatt ttaacaaaat attaacgctt acaattt
//
