LOCUS       sequence-429214-        6795 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCerulean-PCNA in mammalian cells from a retroviral 
            vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6795)
  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 6795)
  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: mCerulean-PCNA-HygroR
FEATURES             Location/Qualifiers
     source          1..6795
                     /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"
     regulatory      1415..1424
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1421..1423,1424..1426,1427..2137)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1466..1487)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1727..1746)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2074..2095
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2992..3012)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2997..3326
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3177..3312
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3400..4425
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3400..4425
                     /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     4942..4961
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             5223..5692
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(join(5807..6598,6599..6667))
                     /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     6430..6449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6668..6772)
                     /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 ggtcgccacc atggtgagca agggcgagga
     1441 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1501 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     1561 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgacctg
     1621 gggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1681 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1741 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1801 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaacgccat
     1861 cagcgacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     1921 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     1981 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa
     2041 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     2101 cgccgggatc actctcggca tggacgagct gtacaagtcc ggagaagggc aagggcaagg
     2161 gcaagggccg ggccgcggct acgcgtatcg atccatgttc gaggcgcgcc tggtccaggg
     2221 ctccatcctc aagaaggtgt tggaggcact caaggacctc atcaacgagg cctgctggga
     2281 tattagctcc agcggtgtaa acctgcagag catggactcg tcccacgtct ctttggtgca
     2341 gctcaccctg cggtctgagg gcttcgacac ctaccgctgc gaccgcaacc tggccatggg
     2401 cgtgaacctc accagtatgt ccaaaatact aaaatgcgcc ggcaatgaag atatcattac
     2461 actaagggcc gaagataacg cggatacctt ggcgctagta tttgaagcac caaaccagga
     2521 gaaagtttca gactatgaaa tgaagttgat ggatttagat gttgaacaac ttggaattcc
     2581 agaacaggag tacagctgtg tagtaaagat gccttctggt gaatttgcac gtatatgccg
     2641 agatctcagc catattggag atgctgttgt aatttcctgt gcaaaagacg gagtgaaatt
     2701 ttctgcaagt ggagaacttg gaaatggaaa cattaaattg tcacagacaa gtaatgtcga
     2761 taaagaggag gaagctgtta ccatagagat gaatgaacca gttcaactaa cttttgcact
     2821 gaggtacctg aacttcttta caaaagccac tccactctct tcaacggtga cactcagtat
     2881 gtctgcagat gtaccccttg ttgtagagta taaaattgcg gatatgggac acttaaaata
     2941 ctacttggct cccaagatcg aggatgaaga aggatcttag gtcgaccctg tggaatgtgt
     3001 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3061 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3121 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3181 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3241 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3301 tttttggagg cctaggcttt tgcaaaaagc ttgccagctg gggcgccctc tggtaaggtt
     3361 gggaagccct gcagatctga tcccgggggg caatgagata tgaaaaagcc tgaactcacc
     3421 gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag
     3481 ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc
     3541 ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt
     3601 gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaatttag cgagagcctg
     3661 acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa
     3721 ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt
     3781 agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg
     3841 cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac
     3901 gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac
     3961 tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac
     4021 aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac
     4081 gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc
     4141 tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc
     4201 cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct
     4261 tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca
     4321 caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat
     4381 agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagagtag atgccgaccg
     4441 ggatctatcg atgcggcggg tgtggtggtt acgcgtaatc tgctgcttgc aaacaaaaaa
     4501 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4561 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4621 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4681 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4741 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4801 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4861 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4921 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4981 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5041 aaacgccagc aacgcatcga taaaataaaa gattttattt agtctccaga aaaagggggg
     5101 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
     5161 atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac
     5221 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     5281 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     5341 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
     5401 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     5461 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     5521 ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg
     5581 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     5641 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt cacatgcagc
     5701 atgtatcaaa attaatttgg ttttttttct taagtattta cattaaatgg ccatagtgcg
     5761 gccgcaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5821 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5881 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5941 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6001 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6061 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6121 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6181 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6241 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6301 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6361 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6421 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6481 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6541 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6601 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6661 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6721 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6781 cccgaaaagt gccac
//
