LOCUS       sequence-216902-        6049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  vector encoding a fusion protein between CDK4 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: CDK4-3XFlag-pBabe
FEATURES             Location/Qualifiers
     source          1..6049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             33..449
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     379..401
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     417..433
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             466..531
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1470..1490)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1475..1804
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1655..1790
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1814..2413
                     /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(1814..1833)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2310..2330
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2634..3103
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3233..3250)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3404..3992)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3484..3503)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4152..4943,4944..5012))
                     /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     4775..4794
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5013..5117)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             5148..5617
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5682..5881
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt
       61 tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca
      121 agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc
      181 cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac
      241 ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt
      301 ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc
      361 catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt
      421 atccagccct cactccttct ctaggcgccg gccggatcca ccatggacta caaagaccat
      481 gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa ggctacctct
      541 cgatatgagc cagtggctga aattggtgtc ggtgcctatg ggacagtgta caaggcccgt
      601 gatccccaca gtggccactt tgtggccctc aagagtgtga gagtccccaa tggaggagga
      661 ggtggaggag gccttcccat cagcacagtt cgtgaggtgg ctttactgag gcgactggag
      721 gcttttgagc atcccaatgt tgtccggctg atggacgtct gtgccacatc ccgaactgac
      781 cgggagatca aggtaaccct ggtgtttgag catgtagacc aggacctaag gacatatctg
      841 gacaaggcac ccccaccagg cttgccagcc gaaacgatca aggatctgat gcgccagttt
      901 ctaagaggcc tagatttcct tcatgccaat tgcatcgttc accgagatct gaagccagag
      961 aacattctgg tgacaagtgg tggaacagtc aagctggctg actttggcct ggccagaatc
     1021 tacagctacc agatggcact tacacccgtg gttgttacac tctggtaccg agctcccgaa
     1081 gttcttctgc agtccacata tgcaacacct gtggacatgt ggagtgttgg ctgtatcttt
     1141 gcagagatgt ttcgtcgaaa gcctctcttc tgtggaaact ctgaagccga ccagttgggc
     1201 aaaatctttg acctgattgg gctgcctcca gaggatgact ggcctcgaga tgtatccctg
     1261 ccccgtggag cctttccccc cagagggccc cgcccagtgc agtcggtggt acctgagatg
     1321 gaggagtcgg gagcacagct gctgctggaa atgctgactt ttaacccaca caagcgaatc
     1381 tctgcctttc gagctctgca gcactcttat ctacataagg atgaaggtaa tccggagtga
     1441 gaattcgcca gcacagtggt cgaccctgtg gaatgtgtgt cagttagggt gtggaaagtc
     1501 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     1561 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     1621 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     1681 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     1741 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     1801 caaaaagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     1861 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     1921 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     1981 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     2041 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     2101 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     2161 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     2221 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     2281 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     2341 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     2401 gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc
     2461 ccatgcatcg ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga
     2521 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa
     2581 tacataactg agaatagaga agttcagatc aaggtcagga acagatggaa cagctgaata
     2641 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     2701 tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     2761 agggccaaga acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca
     2821 tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac
     2881 caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag
     2941 cccacaaccc ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt
     3001 gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg
     3061 gtctcctctg agtgattgac tacccgtcag cgggggtctt tcacatgcag catgtatcaa
     3121 aattaatttg gttttttttc ttaagtattt acattaaatg gccatagttg cattaatgaa
     3181 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3241 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3301 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3361 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3421 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3481 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3541 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3601 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3661 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3721 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3781 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3841 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3901 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3961 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4021 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4081 ggatcttcac ctagatcctt ttgcggccgc aaatcaatct aaagtatata tgagtaaact
     4141 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4201 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4261 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4321 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4381 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4441 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4501 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4561 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4621 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4681 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4741 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4801 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4861 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4921 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4981 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5041 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5101 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcagcctg aatatgggcc
     5161 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
     5221 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     5281 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
     5341 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     5401 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     5461 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
     5521 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     5581 tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg
     5641 gagacccctg cccagggacc accgacccac caccgggagg caagctggcc agcaacttat
     5701 ctgtgtctgt ccgattgtct agtgtctatg actgatttta tgcgcctgcg tcggtactag
     5761 ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc
     5821 ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga
     5881 ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag
     5941 aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc
     6001 gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtc
//
