LOCUS       sequence-429213-        6756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mPlum-PCNA in mammalian cells from a retroviral vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6756)
  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 6756)
  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: mPlum-PCNA-HygroR
FEATURES             Location/Qualifiers
     source          1..6756
                     /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"
     regulatory      1370..1379
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1388..1408
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1421..2098
                     /codon_start=1
                     /product="far-red monomeric derivative of DsRed fluorescent
                     protein (Wang et al., 2004)"
                     /label=mPlum
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKEHMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     RLKVTKGGPLPFAWDILSPQIMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGEFIYKVKVRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEMK
                     MRLRLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRH
                     STGA"
     primer_bind     1822..1841
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2016..2039
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2953..2973)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2958..3287
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3138..3273
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3361..4386
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3361..4386
                     /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"
     primer_bind     4903..4922
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             5184..5653
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(join(5768..6559,6560..6628))
                     /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     6391..6410
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6629..6733)
                     /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 ccggatcccg ccaccatggg
     1381 agcatcacca aaaaagaaga gaaaggtacc ggtggctacc atggtgagca agggcgagga
     1441 ggtcatcaag gagttcatgc gcttcaagga gcacatggag ggctccgtga acggccacga
     1501 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaggct
     1561 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagatcat
     1621 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     1681 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     1741 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga aggtgcgcgg
     1801 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     1861 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatgaaga tgaggctgag
     1921 gctgaaggac ggcggccact acgacgccga ggtcaagacc acctacatgg ccaagaagcc
     1981 cgtgcagctg cccggcgcct acaagaccga catcaagctg gacatcacct cccacaacga
     2041 ggactacacc atcgtggaac agtacgagcg cgccgagggc cgccactcca ccggcgcctc
     2101 cggagaaggg caagggcaag ggcaagggcc gggccgcggc tacgcgtatc gatccatgtt
     2161 cgaggcgcgc ctggtccagg gctccatcct caagaaggtg ttggaggcac tcaaggacct
     2221 catcaacgag gcctgctggg atattagctc cagcggtgta aacctgcaga gcatggactc
     2281 gtcccacgtc tctttggtgc agctcaccct gcggtctgag ggcttcgaca cctaccgctg
     2341 cgaccgcaac ctggccatgg gcgtgaacct caccagtatg tccaaaatac taaaatgcgc
     2401 cggcaatgaa gatatcatta cactaagggc cgaagataac gcggatacct tggcgctagt
     2461 atttgaagca ccaaaccagg agaaagtttc agactatgaa atgaagttga tggatttaga
     2521 tgttgaacaa cttggaattc cagaacagga gtacagctgt gtagtaaaga tgccttctgg
     2581 tgaatttgca cgtatatgcc gagatctcag ccatattgga gatgctgttg taatttcctg
     2641 tgcaaaagac ggagtgaaat tttctgcaag tggagaactt ggaaatggaa acattaaatt
     2701 gtcacagaca agtaatgtcg ataaagagga ggaagctgtt accatagaga tgaatgaacc
     2761 agttcaacta acttttgcac tgaggtacct gaacttcttt acaaaagcca ctccactctc
     2821 ttcaacggtg acactcagta tgtctgcaga tgtacccctt gttgtagagt ataaaattgc
     2881 ggatatggga cacttaaaat actacttggc tcccaagatc gaggatgaag aaggatctta
     2941 ggtcgaccct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3001 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3061 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3121 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3181 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3241 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttgccagct
     3301 ggggcgccct ctggtaaggt tgggaagccc tgcagatctg atcccggggg gcaatgagat
     3361 atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
     3421 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
     3481 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
     3541 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt
     3601 ggggaattta gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg
     3661 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat
     3721 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
     3781 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat
     3841 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag
     3901 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc
     3961 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg
     4021 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct
     4081 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg
     4141 cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
     4201 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
     4261 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc
     4321 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag
     4381 gaatagagta gatgccgacc gggatctatc gatgcggcgg gtgtggtggt tacgcgtaat
     4441 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4501 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4561 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4621 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4681 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4741 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4801 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4861 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4921 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4981 aggggggcgg agcctatgga aaaacgccag caacgcatcg ataaaataaa agattttatt
     5041 tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta
     5101 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga agttcagatc
     5161 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
     5221 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
     5281 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
     5341 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     5401 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     5461 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc
     5521 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     5581 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     5641 cgggggtctt tcacatgcag catgtatcaa aattaatttg gttttttttc ttaagtattt
     5701 acattaaatg gccatagtgc ggccgcaaat caatctaaag tatatatgag taaacttggt
     5761 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5821 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5881 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5941 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6001 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6061 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6121 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6181 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6241 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6301 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6361 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6421 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6481 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6541 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6601 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6661 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6721 taggggttcc gcgcacattt ccccgaaaag tgccac
//
