LOCUS       sequence-364834-        9049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector to co-express human MDH1 with 3xFLAG tag and human
            ME1 with HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9049)
  AUTHORS   Igelmann S, Lessard F, Uchenunu O, Bouchard J, Fernandez-Ruiz A, 
            Rowell MC, Lopes-Paciencia S, Papadopoli D, Fouillen A, Ponce KJ, 
            Huot G, Mignacca L, Benfdil M, Kalegari P, Wahba HM, Pencik J, Vuong
            N, Quenneville J, Guillon J, Bourdeau V, Hulea L, Gagnon E, Kenner 
            L, Moriggl R, Nanci A, Pollak MN, Omichinski JG, Topisirovic I, 
            Ferbeyre G
  TITLE     A hydride transfer complex reprograms NAD metabolism and bypasses 
            senescence.
  JOURNAL   Mol Cell. 2021 Sep 16;81(18):3848-3865.e19. doi: 
            10.1016/j.molcel.2021.08.028.
  PUBMED    34547241
REFERENCE   2  (bases 1 to 9049)
  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: 
            "10.1016/j.molcel.2021.08"; journalName: "Mol Cell"; date: 
            "2021-09-16- 16"; volume: "81"; issue: "18"; pages: "3848-3865.e19"
COMMENT     Sequence Label: pLPC-puromycin-binary-MDH1-3xFLAG-ME1- HA
FEATURES             Location/Qualifiers
     source          1..9049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             942..1358
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1288..1310
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1326..1342
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1376..1975
                     /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(1376..1395)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1872..1892
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2064..2083)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2255..2634
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2635..2838
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2789..2809
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2883..2902
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2883..2901
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3943..4008
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(4197..4214)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4424..4443
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6321..6347
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     LTR             6543..7012
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(7142..7159)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7313..7901)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7393..7412)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8061..8852,8853..8921))
                     /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     8684..8703
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8922..9026)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
      901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
      961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatcagc ttaccatgac
     1381 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     1441 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     1501 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     1561 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     1621 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     1681 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     1741 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     1801 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     1861 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     1921 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacgccc
     1981 gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat cggatctccc
     2041 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     2101 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     2161 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     2221 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
     2281 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     2341 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     2401 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     2461 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     2521 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     2581 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     2641 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     2701 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     2761 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     2821 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
     2881 attaatacga ctcactatag ggagacccaa gcttggtacc gagctcggat ccccaccatg
     2941 tctgaaccaa tcagagtcct tgtgactgga gcagctggtc aaattgcata ttcactgctg
     3001 tacagtattg gaaatggatc tgtctttggt aaagatcagc ctataattct tgtgctgttg
     3061 gatatcaccc ccatgatggg tgtcctggac ggtgtcctaa tggaactgca agactgtgcc
     3121 cttcccctcc tgaaagatgt catcgcaaca gataaagaag acgttgcctt caaagacctg
     3181 gatgtggcca ttcttgtggg ctccatgcca agaagggaag gcatggagag aaaagattta
     3241 ctgaaagcaa atgtgaaaat cttcaaatcc cagggtgcag ccttagataa atacgccaag
     3301 aagtcagtta aggttattgt tgtgggtaat ccagccaata ccaactgcct gactgcttcc
     3361 aagtcagctc catccatccc caaggagaac ttcagttgct tgactcgttt ggatcacaac
     3421 cgagctaaag ctcaaattgc tcttaaactt ggtgtgactg ctaatgatgt aaagaatgtc
     3481 attatctggg gaaaccattc ctcgactcag tatccagatg tcaaccatgc caaggtgaaa
     3541 ttgcaaggaa aggaagttgg tgtttatgaa gctctgaaag atgacagctg gctcaaggga
     3601 gaatttgtca cgactgtgca gcagcgtggc gctgctgtca tcaaggctcg aaaactatcc
     3661 agtgccatgt ctgctgcaaa agccatctgt gaccacgtca gggacatctg gtttggaacc
     3721 ccagagggag agtttgtgtc catgggtgtt atctctgatg gcaactccta tggtgttcct
     3781 gatgatctgc tctactcatt ccctgttgta atcaagaata agacctggaa gtttgttgaa
     3841 ggtctcccta ttaatgattt ctcacgtgag aagatggatc ttactgcaaa ggaactgaca
     3901 gaagaaaaag aaagtgcttt tgaatttctt tcctctgcca tggactacaa agaccatgac
     3961 ggtgattata aagatcatga catcgattac aaggatgacg atgacaagtg agaattctgc
     4021 agatatccat cacactggcg gccgctcgag taacgttact ggccgaagcc gcttggaata
     4081 aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt
     4141 gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct
     4201 cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc
     4261 ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga
     4321 caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc
     4381 ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt
     4441 attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg
     4501 gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc
     4561 gaaccacggg gacgtggttt tcctttgaaa aacacgatag atccatggag cccgaagccc
     4621 cccgtcgccg ccacacccat cagcgcggct acctgctgac acggaaccct cacctcaaca
     4681 aggacttggc ctttaccctg gaagagagac agcaattgaa cattcatgga ttgttgccac
     4741 cttccttcaa cagtcaggag atccaggttc ttagagtagt aaaaaatttc gagcatctga
     4801 actctgactt tgacaggtat cttctcttaa tggatctcca agatagaaat gaaaaactct
     4861 tttatagagt gctgacatct gacattgaga aattcatgcc tattgtttat actcccactg
     4921 tgggtctggc ttgccaacaa tatagtttgg tgtttcggaa gccaagaggt ctctttatta
     4981 ctatccacga tcgagggcat attgcttcag ttctcaatgc atggccagaa gatgtcatca
     5041 aggccattgt ggtgactgat ggagagcgta ttcttggctt gggagacctt ggctgtaatg
     5101 gaatgggcat ccctgtgggt aaattggctc tatatacagc ttgcggaggg atgaatcctc
     5161 aagaatgtct gcctgtcatt ctggatgtgg gaaccgaaaa tgaggagtta cttaaagatc
     5221 cactctacat tggactacgg cagagaagag taagaggttc tgaatatgat gattttttgg
     5281 acgaattcat ggaggcagtt tcttccaagt atggcatgaa ttgccttatt cagtttgaag
     5341 attttgccaa tgtgaatgca tttcgtctcc tgaacaagta tcgaaaccag tattgcacat
     5401 tcaatgatga tattcaagga acagcatctg ttgcagttgc aggtctcctt gcagctcttc
     5461 gaataaccaa gaacaaactg tctgatcaaa caatactatt ccaaggagct ggagaggctg
     5521 ccctagggat tgcacacctg attgtgatgg ccttggaaaa agaaggttta ccaaaagaga
     5581 aagccatcaa aaagatatgg ctggttgatt caaaaggatt aatagttaag ggacgtgctt
     5641 ccttaacaca agagaaagag aagtttgccc atgaacatga agaaatgaag aacctagaag
     5701 ccattgttca agaaataaaa ccaactgccc tcataggagt tgctgcaatt ggtggtgcat
     5761 tctcagaaca aattctcaaa gatatggctg ccttcaatga acggcctatt atttttgctt
     5821 tgagtaatcc aactagcaaa gcagaatgtt ctgcagagca gtgctacaaa ataaccaagg
     5881 gacgtgcaat ttttgccagt ggcagtcctt ttgatccagt cactcttcca aatggacaga
     5941 ccctatatcc tggccaaggc aacaattcct atgtgttccc tggagttgct cttggtgttg
     6001 tggcgtgtgg attgaggcag atcacagata atattttcct cactactgct gaggttatag
     6061 ctcagcaagt gtcagataaa cacttggaag agggtcggct ttatcctcct ttgaatacca
     6121 ttagagatgt ttctctgaaa attgcagaaa agattgtgaa agatgcatac caagaaaaga
     6181 cagccacagt ttatcctgaa ccgcaaaaca aagaagcatt tgtccgctcc cagatgtata
     6241 gtactgatta tgaccagatt ctacctgatt gttattcttg gcctgaagag gtgcagaaaa
     6301 tacagaccaa agttgaccag tacccatacg atgttccaga ttacgcttag ctcgactcga
     6361 gcatgcatct agagggccga taaaataaaa gattttattt agtctccaga aaaagggggg
     6421 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
     6481 atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac
     6541 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     6601 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     6661 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
     6721 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     6781 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     6841 ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg
     6901 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     6961 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt cacatgcagc
     7021 atgtatcaaa attaatttgg ttttttttct taagtattta cattaaatgg ccatagttgc
     7081 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     7141 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     7201 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     7261 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7321 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7381 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7441 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     7501 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     7561 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7621 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     7681 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7741 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7801 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7861 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7921 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7981 tatcaaaaag gatcttcacc tagatccttt tgcggccgca aatcaatcta aagtatatat
     8041 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     8101 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     8161 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     8221 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     8281 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     8341 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     8401 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     8461 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     8521 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     8581 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     8641 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     8701 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8761 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8821 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8881 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8941 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     9001 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccac
//
