LOCUS       sequence-426803-        8595 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry-POLB in mammalian cells from a retroviral vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8595)
  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 8595)
  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-POLB
FEATURES             Location/Qualifiers
     source          1..8595
                     /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        3990..4489
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4025..4048)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4422..4441
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4510..5109
                     /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(4510..4529)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5006..5026
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5294..5726
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5829..5851)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5987..6007)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5992..6321
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6172..6307
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6343..6362
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6443..6460)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6614..7202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6694..6713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7373..8164,8165..8233))
                     /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     7996..8015
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8234..8338)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8406..8424
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8524..8595,1..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 agatctcgag ctcaagcttc gaattctgca
     2941 gtcgacagca aacggaaggc gccgcaggag actctcaacg ggggaatcac cgacatgctc
     3001 acagaactcg caaactttga gaagaacgtg agccaagcta tccacaagta caatgcttac
     3061 agaaaagcag catctgttat agcaaaatac ccacacaaaa taaagagtgg agctgaagct
     3121 aagaaattgc ctggagtagg aacaaaaatt gctgaaaaga ttgatgagtt tttagcaact
     3181 ggaaaattac gtaaactgga aaagattcgg caggatgata cgagttcatc catcaatttc
     3241 ctgactcgag ttagtggcat tggtccatct gctgcaagga agtttgtaga tgaaggaatt
     3301 aaaacactag aagatctcag aaaaaatgaa gataaattga accatcatca gcgaattggg
     3361 ctgaaatatt ttggggactt tgaaaaaaga attcctcgtg aagagatgtt acaaatgcaa
     3421 gatattgtac taaatgaagt taaaaaagtg gattctgaat acattgctac agtctgtggc
     3481 agtttcagaa gaggtgcaga gtccagtggt gacatggatg ttctcctgac ccatcccagc
     3541 ttcacttcag aatcaaccaa acagccaaaa ctgttacatc aggttgtgga gcagttacaa
     3601 aaggttcatt ttatcacaga taccctgtca aagggtgaga caaagttcat gggtgtttgc
     3661 cagcttccca gtaaaaatga tgaaaaagaa tatccacaca gaagaattga tatcaggttg
     3721 atacccaaag atcagtatta ctgtggtgtt ctctatttca ctgggagtga tattttcaat
     3781 aagaatatga gggctcatgc cctagaaaag ggtttcacaa tcaatgagta caccatccgt
     3841 cccttgggag tcactggagt tgcaggagaa cccctgccag tggatagtga aaaagacatc
     3901 tttgattaca tccagtggaa ataccgggaa cccaaggacc ggagcgaatg aggatccacc
     3961 ggatctagat aactgatcat aattctaccg ggtaggggag gcgcttttcc caaggcagtc
     4021 tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc
     4081 tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt
     4141 cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc
     4201 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac
     4261 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt
     4321 cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct
     4381 caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa
     4441 gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagccc
     4501 aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     4561 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     4621 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     4681 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     4741 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     4801 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     4861 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     4921 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     4981 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     5041 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     5101 ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc acgaccccat
     5161 gcatcggcgt ctcgatcgag atatcagtgg tccaggctct agttttgact caacaatatc
     5221 accagctgaa gcctatagag tacgagccat agataaaata aaagatttta tttagtctcc
     5281 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctacta gcttaagtaa
     5341 cgccattttg caaggcatgg aaaaatacat aactgagaat agagaagttc agatcaaggt
     5401 caggaacaga tggaacaggg tcgatcgacc ctagagaacc atcagatgtt tccagggtgc
     5461 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     5521 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     5581 ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt
     5641 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     5701 ctacccgtca gcgggggtct ttcatttggg ggctcgtccg ggatcgggag acccctgccc
     5761 agggaccacc gacccaccac cgggaggtaa gctggctgcc tcgcgcgttt cggtgatgac
     5821 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     5881 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca
     5941 gccatgaccc agtcacgtag cgatagcgga gtgtagatcc ggctgtggaa tgtgtgtcag
     6001 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     6061 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     6121 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     6181 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     6241 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     6301 ggaggcctag gcttttgcaa aaagcttact ggcttaacta tgcggcatca gagcagattg
     6361 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     6421 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6481 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6541 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6601 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6661 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6721 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6781 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6841 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6901 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6961 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7021 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     7081 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7141 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7201 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7261 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7321 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7381 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7441 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7501 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7561 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7621 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7681 ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7741 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7801 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7861 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7921 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7981 cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8041 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8101 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8161 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8221 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8281 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8341 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     8401 ataaaaatag gcgtatcacg aggccctttc gtcttcaaga attagcttgg ccattgcata
     8461 cgttgtatcc atatcataat atgtacattt atattggctc atgtccaaca ttaccgccat
     8521 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     8581 gcccatatat ggagt
//
