LOCUS       sequence-237893-        6049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  vector encoding a fusion protein between a CDK4 mutant (K35M) and 3 
            Flag tags.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6049)
  AUTHORS   Acevedo M, Vernier M, Mignacca L, Lessard F, Huot G, Moiseeva O, 
            Bourdeau V, Ferbeyre G
  TITLE     A CDK4/6-Dependent Epigenetic Mechanism Protects Cancer Cells from 
            PML-induced Senescence.
  JOURNAL   Cancer Res. 2016 Jun 1;76(11):3252-64. doi: 
            10.1158/0008-5472.CAN-15-2347. Epub 2016 Mar 29.
  PUBMED    27206849
REFERENCE   2  (bases 1 to 6049)
  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: 
            "10.1158/0008-5472.CAN-15-2347"; journalName: "Cancer Res"; date: 
            "2016-06-1- 1"; volume: "76"; issue: "11"; pages: "3252-64"
COMMENT     Sequence Label: CDK4mut(K35M)-3XFlag-pBabe
FEATURES             Location/Qualifiers
     source          1..6049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(939..959)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        944..1273
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1124..1259
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1283..1882
                     /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(1283..1302)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1779..1799
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2103..2572
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2702..2719)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2873..3461)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2953..2972)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3621..4412,4413..4481))
                     /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     4244..4263
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4482..4586)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             4617..5086
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5151..5350
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5551..5967
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5897..5919
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5935..5951
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             5984..6049
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 gctacctctc gatatgagcc agtggctgaa attggtgtcg gtgcctatgg gacagtgtac
       61 aaggcccgtg atccccacag tggccacttt gtggccctca tgagtgtgag agtccccaat
      121 ggaggaggag gtggaggagg ccttcccatc agcacagttc gtgaggtggc tttactgagg
      181 cgactggagg cttttgagca tcccaatgtt gtccggctga tggacgtctg tgccacatcc
      241 cgaactgacc gggagatcaa ggtaaccctg gtgtttgagc atgtagacca ggacctaagg
      301 acatatctgg acaaggcacc cccaccaggc ttgccagccg aaacgatcaa ggatctgatg
      361 cgccagtttc taagaggcct agatttcctt catgccaatt gcatcgttca ccgagatctg
      421 aagccagaga acattctggt gacaagtggt ggaacagtca agctggctga ctttggcctg
      481 gccagaatct acagctacca gatggcactt acacccgtgg ttgttacact ctggtaccga
      541 gctcccgaag ttcttctgca gtccacatat gcaacacctg tggacatgtg gagtgttggc
      601 tgtatctttg cagagatgtt tcgtcgaaag cctctcttct gtggaaactc tgaagccgac
      661 cagttgggca aaatctttga cctgattggg ctgcctccag aggatgactg gcctcgagat
      721 gtatccctgc cccgtggagc ctttcccccc agagggcccc gcccagtgca gtcggtggta
      781 cctgagatgg aggagtcggg agcacagctg ctgctggaaa tgctgacttt taacccacac
      841 aagcgaatct ctgcctttcg agctctgcag cactcttatc tacataagga tgaaggtaat
      901 ccggagtgag aattcgccag cacagtggtc gaccctgtgg aatgtgtgtc agttagggtg
      961 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     1021 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     1081 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     1141 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     1201 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     1261 aggcttttgc aaaaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     1321 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     1381 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     1441 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     1501 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     1561 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     1621 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     1681 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     1741 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     1801 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     1861 gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag
     1921 cgcacgaccc catgcatcga taaaataaaa gattttattt agtctccaga aaaagggggg
     1981 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
     2041 atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac
     2101 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     2161 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     2221 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
     2281 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     2341 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     2401 ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg
     2461 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     2521 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt cacatgcagc
     2581 atgtatcaaa attaatttgg ttttttttct taagtattta cattaaatgg ccatagttgc
     2641 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2701 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2761 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2821 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2881 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2941 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3001 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3061 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3121 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3181 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3241 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3301 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3361 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3421 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3481 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3541 tatcaaaaag gatcttcacc tagatccttt tgcggccgca aatcaatcta aagtatatat
     3601 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3661 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3721 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3781 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3841 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3901 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3961 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4021 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4081 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4141 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4201 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4261 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4321 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4381 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4441 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4501 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4561 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgcagcctga
     4621 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     4681 agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
     4741 ctcagggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagagaa
     4801 ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact
     4861 aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa
     4921 gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc
     4981 cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg
     5041 agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatttg ggggctcgtc
     5101 cgggatcggg agacccctgc ccagggacca ccgacccacc accgggaggc aagctggcca
     5161 gcaacttatc tgtgtctgtc cgattgtcta gtgtctatga ctgattttat gcgcctgcgt
     5221 cggtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac tgacgagttc
     5281 tgaacacccg gccgcaaccc tgggagacgt cccagggact ttgggggccg tttttgtggc
     5341 ccgacctgag gaagggagtc gatgtggaat ccgaccccgt caggatatgt ggttctggta
     5401 ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc ggtttggaac
     5461 cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg
     5521 actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc cttaagtttg
     5581 accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag
     5641 aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg
     5701 cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct
     5761 ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt
     5821 gacccccctc cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca
     5881 tccgccccgt ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat
     5941 ccagccctca ctccttctct aggcgccggc cggatccacc atggactaca aagaccatga
     6001 cggtgattat aaagatcatg acatcgatta caaggatgac gatgacaag
//
