LOCUS       sequence-426796-        8625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry-APTX in mammalian cells from a retroviral vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8625)
  AUTHORS   Rona G, Miwatani-Minter B, Zhang Q, Goldberg HV, Kerzhnerman MA, 
            Howard JB, Simoneschi D, Lane E, Hobbs JW, Sassani E, Wang AA, 
            Keegan S, Laverty DJ, Piett CG, Pongor LS, Xu ML, Andrade J, Thomas 
            A, Sicinski P, Askenazi M, Ueberheide B, Fenyo D, Nagel ZD, Pagano M
  TITLE     CDK-independent role of D-type cyclins in regulating DNA mismatch 
            repair.
  JOURNAL   Mol Cell. 2024 Feb 29:S1097-2765(24)00129-1. doi: 
            10.1016/j.molcel.2024.02.010.
  PUBMED    38458201
REFERENCE   2  (bases 1 to 8625)
  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"; journalName: "Mol Cell. 
            2024 Feb 29:S1097-2765(24)00129-1. doi: 
            10.1016/j.molcel.2024.02.010."
COMMENT     Sequence Label: mCherry-APTX
FEATURES             Location/Qualifiers
     source          1..8625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        309..512
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             513..688
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    751..950
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1151..1567
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1497..1519
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1535..1551
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1642..1945
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1946..2149
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      2188..2197
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2194..2901
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2339..2357)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2607..2626
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        4020..4519
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4055..4078)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4452..4471
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4540..5139
                     /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(4540..4559)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5036..5056
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5324..5756
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5859..5881)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6017..6037)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6022..6351
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6202..6337
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6373..6392
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6473..6490)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6644..7232)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6724..6743)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7403..8194,8195..8263))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8026..8045
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8264..8368)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8436..8454
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8554..8625,1..308)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
       61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      301 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      361 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      421 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      481 gtgtacggtg ggaggtctat ataagcagag ctcaataaaa gagcccacaa cccctcactc
      541 ggcgcgccag tcttccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc
      601 ttgctgtttg catccgaatc gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
      661 tgactaccca cgacgggggt ctttcatttg ggggctcgtc cgggatttgg agacccctgc
      721 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc
      781 cgattgtcta gtgtctatgt ttgatgttat gcgcctgcgt ctgtactagt tagctaacta
      841 gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc
      901 tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc
      961 gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca
     1021 gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct
     1081 gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc
     1141 tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga
     1201 tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt
     1261 ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg
     1321 agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga
     1381 ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa
     1441 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct
     1501 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct
     1561 aggcgccgga attgaagatc atagttatta atagtaatca attacggggt cattagttca
     1621 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     1681 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     1741 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     1801 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     1861 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     1921 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     1981 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     2041 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     2101 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     2161 ccgtcagatc cgctagcgct accggtcgcc accatggtga gcaagggcga ggaggataac
     2221 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     2281 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     2341 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     2401 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     2461 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     2521 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     2581 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     2641 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     2701 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     2761 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     2821 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     2881 ggcatggacg agctgtacaa gtccggactc agatctggag gcggaggaag catgatgcgg
     2941 gtgtgctggt tggtgagaca ggacagccgg caccagcgaa tcagacttcc acatttggaa
     3001 gcagttgtga ttgggcgtgg cccagagacc aagatcactg ataagaaatg ttctcgacag
     3061 caagtacagt tgaaagcaga gtgtaacaag ggatatgtca aggtaaagca ggtaggagtc
     3121 aatcccacca gcattgactc agtcgtaatt gggaaggacc aagaggtgaa gctgcagcct
     3181 ggccaggttc tccacatggt gaatgaactt tatccatata ttgtagagtt tgaggaagag
     3241 gcaaagaacc ctggcctgga aacacacagg aagagaaaga gatcaggcaa cagtgattct
     3301 atagaaaggg atgctgctca ggaagctgag gctgggacag ggctggaacc tgggagcaac
     3361 tctggccaat gctctgtgcc cctaaagaag ggaaaagatg cacctatcaa aaaggaatcc
     3421 ctgggccact ggagtcaagg cttgaagatt tctatgcagg accccaaaat gcaggtttac
     3481 aaagatgagc aggtggtggt gataaaggat aaatacccaa aggcccgtta ccattggctg
     3541 gtcttaccgt ggacctccat ttccagtctg aaggctgtgg ccagggaaca ccttgaactc
     3601 cttaagcata tgcacactgt gggggaaaag gtgattgtag attttgctgg gtccagcaaa
     3661 ctccgcttcc gattgggcta ccacgccatt ccgagtatga gccatgtaca tcttcatgtg
     3721 atcagccagg attttgattc tccttgcctt aaaaacaaaa aacattggaa ttctttcaat
     3781 acagaatact tcctagaatc acaagctgtg atcgagatgg tacaagaggc tggtagagta
     3841 actgtccgag atgggatgcc tgagctcttg aagctgcccc ttcgttgtca tgagtgccag
     3901 cagctgctgc cttccattcc tcagctgaaa gaacatctca ggaagcactg gacacagtga
     3961 gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat aattctaccg
     4021 ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg
     4081 cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg
     4141 ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag
     4201 gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg
     4261 tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg
     4321 gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg
     4381 tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc
     4441 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc
     4501 ctcatctccg ggcctttcga cctgcagccc aagcttacca tgaccgagta caagcccacg
     4561 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     4621 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     4681 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     4741 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     4801 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     4861 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     4921 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     4981 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     5041 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     5101 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag
     5161 cgcccgaccg aaaggagcgc acgaccccat gcatcggcgt ctcgatcgag atatcagtgg
     5221 tccaggctct agttttgact caacaatatc accagctgaa gcctatagag tacgagccat
     5281 agataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     5341 gtaggtttgg caagctacta gcttaagtaa cgccattttg caaggcatgg aaaaatacat
     5401 aactgagaat agagaagttc agatcaaggt caggaacaga tggaacaggg tcgatcgacc
     5461 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     5521 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     5581 caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc
     5641 cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt
     5701 tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatttggg
     5761 ggctcgtccg ggatcgggag acccctgccc agggaccacc gacccaccac cgggaggtaa
     5821 gctggctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     5881 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     5941 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     6001 gtgtagatcc ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     6061 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     6121 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     6181 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     6241 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     6301 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttact
     6361 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     6421 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
     6481 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6541 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6601 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6661 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6721 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6781 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6841 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6901 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6961 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7021 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7081 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7141 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7201 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7261 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7321 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7381 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7441 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7501 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7561 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7621 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7681 cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct
     7741 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7801 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7861 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7921 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7981 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
     8041 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8101 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8161 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8221 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8281 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8341 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     8401 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     8461 gtcttcaaga attagcttgg ccattgcata cgttgtatcc atatcataat atgtacattt
     8521 atattggctc atgtccaaca ttaccgccat gttgacattg attattgact agttattaat
     8581 agtaatcaat tacggggtca ttagttcata gcccatatat ggagt
//
