LOCUS       sequence-238518-        7504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses RPS14 protein with a Myc tag in mammalian cells (hygro 
            resistance).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7504)
  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 7504)
  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: MSCV-RPS14(WT)-Myc
FEATURES             Location/Qualifiers
     source          1..7504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1333..1355
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1371..1387
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1924..1953
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        1997..2496
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2032..2055)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2429..2448
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2514..3539
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2514..3539
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             3897..4411
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4573..4589
                     /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(4597..4603,4604..4621,4622..4627))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4642..4663
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4780..4797)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4951..5539)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5031..5050)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5710..6501,6502..6570))
                     /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     6333..6352
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6571..6675)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6743..6761
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6799..6821)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6921..6940
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7134..7156
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7284..7303
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7503..7504,1..515)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatctggatc cggtaccgag gagatctgcc
     1441 gccgcgatcg ccatggcacc tcgaaagggg aaggaaaaga aggaagaaca ggtcatcagc
     1501 ctcggacctc aggtggctga aggagagaat gtatttggtg tctgccatat ctttgcatcc
     1561 ttcaatgaca cttttgtcca tgtcactgat ctttctggca aagaaaccat ctgccgtgtg
     1621 actggtggga tgaaggtaaa ggcagaccga gatgaatcct caccatatgc tgctatgttg
     1681 gctgcccagg atgtggccca gaggtgcaag gagctgggta tcaccgccct acacatcaaa
     1741 ctccgggcca caggaggaaa taggaccaag acccctggac ctggggccca gtcggccctc
     1801 agagcccttg cccgctcggg tatgaagatc gggcggattg aggatgtcac ccccatcccc
     1861 tctgacagca ctcgcaggaa ggggggtcgc cgtggtcgcc gtctgacgcg tacgcggccg
     1921 ctcgagcaga aactcatctc agaagaggat ctggcataag tcgagcaatg catggttaac
     1981 cacaacgaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct
     2041 ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc
     2101 acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct
     2161 actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga
     2221 caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg
     2281 aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct
     2341 cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg
     2401 ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc
     2461 gcgctgttct cctcttcctc atctccgggc ctttcgacct gcatcccgcc accatgaaaa
     2521 agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct
     2581 ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag
     2641 ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg
     2701 tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggaat
     2761 tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc
     2821 tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg gatgcgatcg
     2881 ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc
     2941 aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc
     3001 aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc
     3061 tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca
     3121 atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg
     3181 gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg
     3241 agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc
     3301 gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt
     3361 tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga
     3421 ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag
     3481 aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaataga
     3541 gtagatgccg accgaacaag agctgatttc gagaacgcct cagccagcaa ctcgcgcgag
     3601 cctagcaagg caaatgcgag agaacggcct tacgcttggt ggcacagttc tcgtccacag
     3661 ttcgctaagc tcgctcggct gggtcgcggg agggccggtc gcagtgattc aggcccttct
     3721 ggattgtgtt ggtccccagg gcacgattgt catgcccacg cactcgggtg atctgactga
     3781 tcccgcagat tggagatcgc cgcccgtgcc tgccgattgg gtgcagatcc gtcgacctgc
     3841 agccaagctt atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga
     3901 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     3961 aaatacataa ctgagaatag agaagttcag atcaaggtta ggaacagaga gacagcagaa
     4021 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     4081 gatggtcccc agatgcggtc ccgccctcag cagtttctag agaaccatca gatgtttcca
     4141 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
     4201 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
     4261 actcggcgcg ccagtcctcc gatagactgc gtcgcccggg tacccgtgta tccaataaac
     4321 cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt
     4381 gattgactac ccgtcagcgg gggtctttca tgggtaacag tttcttgaag ttggagaaca
     4441 acattctgag ggtaggagtc gaatattaag taatcctgac tcaattagcc actgttttga
     4501 atccacatac tccaatactc ctgaaatagt tcattatgga cagcgcagaa agagctgggg
     4561 agaattgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4621 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4681 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4741 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4801 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4861 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4921 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4981 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5041 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5101 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5161 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5221 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5281 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5341 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5401 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5461 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5521 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5581 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5641 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5701 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5761 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5821 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5881 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5941 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6001 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6061 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6121 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6181 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6241 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6301 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6361 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6421 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6481 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6541 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6601 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6661 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     6721 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     6781 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     6841 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     6901 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     6961 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     7021 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     7081 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     7141 acgacgttgt aaaacgacgg cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     7201 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     7261 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     7321 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggcgattagt ccaatttgtt
     7381 aaagacagga tatcagtggt ccaggctcta gttttgactc aacaatatca ccagctgaag
     7441 cctatagagt acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg
     7501 ggaa
//
