LOCUS       sequence-238492-        5588 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses RPS14 protein with a Myc tag in mammalian cells (puro 
            resistance).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5588)
  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 5588)
  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: pBABE-RPS14(WT)-Myc
FEATURES             Location/Qualifiers
     source          1..5588
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             513..542
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(560..580)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        565..894
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      745..880
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             904..1503
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(904..923)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1400..1420
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             1724..2193
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2323..2340)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2494..3082)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2574..2593)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3242..4033,4034..4102))
                     /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     3865..3884
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4103..4207)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             4238..4707
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    4772..4971
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5172..5588
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5518..5540
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5556..5572
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 ggccggatcc ggtaccgagg agatctgccg ccgcgatcgc catggcacct cgaaagggga
       61 aggaaaagaa ggaagaacag gtcatcagcc tcggacctca ggtggctgaa ggagagaatg
      121 tatttggtgt ctgccatatc tttgcatcct tcaatgacac ttttgtccat gtcactgatc
      181 tttctggcaa agaaaccatc tgccgtgtga ctggtgggat gaaggtaaag gcagaccgag
      241 atgaatcctc accatatgct gctatgttgg ctgcccagga tgtggcccag aggtgcaagg
      301 agctgggtat caccgcccta cacatcaaac tccgggccac aggaggaaat aggaccaaga
      361 cccctggacc tggggcccag tcggccctca gagcccttgc ccgctcgggt atgaagatcg
      421 ggcggattga ggatgtcacc cccatcccct ctgacagcac tcgcaggaag gggggtcgcc
      481 gtggtcgccg tctgacgcgt acgcggccgc tcgagcagaa actcatctca gaagaggatc
      541 tggcataagt cgaccctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
      601 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
      661 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
      721 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
      781 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
      841 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt
      901 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
      961 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     1021 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     1081 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     1141 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     1201 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     1261 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     1321 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     1381 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     1441 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     1501 tgacgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg
     1561 ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
     1621 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg
     1681 agaatagaga agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac
     1741 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct
     1801 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     1861 acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt
     1921 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
     1981 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
     2041 ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata
     2101 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg
     2161 agtgattgac tacccgtcag cgggggtctt tcacatgcag catgtatcaa aattaatttg
     2221 gttttttttc ttaagtattt acattaaatg gccatagttg cattaatgaa tcggccaacg
     2281 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     2341 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2401 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2461 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2521 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2581 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2641 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2701 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2761 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2821 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2881 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2941 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3001 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3061 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3121 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3181 ctagatcctt ttgcggccgc aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3241 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3301 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3361 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3421 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3481 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3541 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3601 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3661 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3721 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3781 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3841 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3901 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3961 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4021 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4081 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     4141 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     4201 ttccgcgcac atttccccga aaagtgccac ctgcagcctg aatatgggcc aaacaggata
     4261 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat
     4321 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     4381 ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg
     4441 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
     4501 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
     4561 cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct
     4621 cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
     4681 tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg gagacccctg
     4741 cccagggacc accgacccac caccgggagg caagctggcc agcaacttat ctgtgtctgt
     4801 ccgattgtct agtgtctatg actgatttta tgcgcctgcg tcggtactag ttagctaact
     4861 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc
     4921 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt
     4981 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac
     5041 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc
     5101 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt
     5161 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
     5221 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
     5281 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
     5341 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
     5401 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
     5461 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
     5521 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
     5581 taggcgcc
//
