LOCUS       sequence-238565-        5903 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses RPS14 protein with a flag tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5903)
  AUTHORS   Lessard F, Igelmann S, Trahan C, Huot G, Saint-Germain E, Mignacca 
            L, Del Toro N, Lopes-Paciencia S, Le Calve B, Montero M, 
            Deschenes-Simard X, Bury M, Moiseeva O, Rowell MC, Zorca CE, 
            Zenklusen D, Brakier-Gingras L, Bourdeau V, Oeffinger M, Ferbeyre G
  TITLE     Senescence-associated ribosome biogenesis defects contributes to 
            cell cycle arrest through the Rb pathway.
  JOURNAL   Nat Cell Biol. 2018 Jul;20(7):789-799. doi: 
            10.1038/s41556-018-0127-y. Epub 2018 Jun 25.
  PUBMED    29941930
REFERENCE   2  (bases 1 to 5903)
  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: 
            "10.1038/s41556-018-0127-y"; journalName: "Nat Cell Biol"; date: 
            "2018-07"; volume: "20"; issue: "7"; pages: "789-799"
COMMENT     Sequence Label: pcDNA3-FLAG-RPS14(WT)
FEATURES             Location/Qualifiers
     source          1..5903
                     /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"
     primer_bind     249..268
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        249..267
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             305..328
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(842..860)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(843..860)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(880..897)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    886..1110
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1156..1584
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1243..1262)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1453..1474
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1593..1613)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1598..1927
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1778..1913
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1840..1859
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1994..2788
                     /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(2048..2067)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2658..2677
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2962..3083
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2999..3018)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3053..3072
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3132..3148)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3132..3148)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3145..3167)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3156..3172
                     /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(3180..3186,3187..3204,3205..3210))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3225..3246
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3363..3380)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3534..4122)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3614..3633)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4293..5084,5085..5153))
                     /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     4916..4935
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5154..5258)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5333..5352)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5524..5903
                     /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 agctctctgg ctaactagag aacccactgc ttactggctt
      241 atcgaaatta atacgactca ctatagggag acccaagctt ggtaccgagc tcggatccac
      301 catggattac aaggatgacg atgacaaggc acctcgaaag gggaaggaaa agaaggaaga
      361 acaggtcatc agcctcggac ctcaggtggc tgaaggagag aatgtatttg gtgtctgcca
      421 tatctttgca tccttcaatg acacttttgt ccatgtcact gatctttctg gcaaagaaac
      481 catctgccgt gtgactggtg ggatgaaggt aaaggcagac cgagatgaat cctcaccata
      541 tgctgctatg ttggctgccc aggatgtggc ccagaggtgc aaggagctgg gtatcaccgc
      601 cctacacatc aaactccggg ccacaggagg aaataggacc aagacccctg gacctggggc
      661 ccagtcggcc ctcagagccc ttgcccgctc gggtatgaag atcgggcgga ttgaggatgt
      721 cacccccatc ccctctgaca gcactcgcag gaaggggggt cgccgtggtc gccgtctgtg
      781 agaattctgc agatatccat cacactggcg gccgctcgag catgcatcta gagggcccta
      841 ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt
      901 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
      961 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     1021 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     1081 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     1141 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     1201 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     1261 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     1321 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     1381 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     1441 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     1501 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     1561 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     1621 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     1681 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     1741 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     1801 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     1861 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     1921 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     1981 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     2041 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     2101 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     2161 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     2221 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     2281 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     2341 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     2401 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     2461 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     2521 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     2581 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     2641 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     2701 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     2761 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     2821 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     2881 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     2941 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     3001 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3061 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     3121 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3181 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3241 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3301 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3361 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3421 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3481 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3541 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3601 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3661 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3721 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3781 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3841 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3901 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3961 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4021 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4081 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4141 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4201 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4261 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4321 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4381 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4441 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4501 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4561 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4621 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4681 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4741 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4801 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4861 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4921 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4981 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5041 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5101 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5161 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5221 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5281 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     5341 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     5401 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     5461 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     5521 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     5581 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     5641 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     5701 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     5761 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     5821 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     5881 tattagtcat cgctattacc atg
//
