LOCUS       sequence-429302-        6656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mKO2-CDK4 in mammalian cells from a retroviral vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6656)
  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 6656)
  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: mKO2-CDK4
FEATURES             Location/Qualifiers
     source          1..6656
                     /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             1376..2029
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     primer_bind     complement(2986..3006)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2991..3320
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3171..3306
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3330..3929
                     /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(3330..3349)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3826..3846
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4150..4619
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4749..4766)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4920..5508)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5000..5019)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5668..6459,6460..6528))
                     /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     6291..6310
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6529..6633)
                     /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 ccaccatggt
     1381 gagtgtgatt aaaccagaga tgaagatgag gtactacatg gacggctccg tcaatgggca
     1441 tgagttcaca attgaaggtg aaggcacagg cagaccttac gagggacatc aagagatgac
     1501 actacgcgtc acaatggccg agggcgggcc aatgcctttc gcgtttgact tagtgtcaca
     1561 cgtgttctgt tacggccaca gagtatttac taaatatcca gaagagatac cagactattt
     1621 caaacaagca tttcctgaag gcctgtcatg ggaaaggtcg ttggagttcg aagatggtgg
     1681 gtccgcttca gtcagtgcgc atataagcct tagaggaaac accttctacc acaaatccaa
     1741 atttactggg gttaactttc ctgccgatgg tcctatcatg caaaaccaaa gtgttgattg
     1801 ggagccatca accgagaaaa ttactgccag cgacggagtt ctgaagggtg atgttacgat
     1861 gtacctaaaa cttgaaggag gcggcaatca caaatgccaa atgaagacta cttacaaggc
     1921 ggcaaaagag attcttgaaa tgccaggaga ccattacatc ggccatcgcc tcgtcaggaa
     1981 aaccgaaggc aacattactg agcaggtaga agatgcagta gctcattccg gactcagatc
     2041 tcgagctcaa gcttcgaatt caccagctac ctctcgatat gagccagtgg ctgaaattgg
     2101 tgtcggtgcc tatgggacag tgtacaaggc ccgtgatccc cacagtggcc actttgtggc
     2161 cctcaagagt gtgagagtcc ccaatggagg aggaggtgga ggaggccttc ccatcagcac
     2221 agttcgtgag gtggctttac tgaggcgact ggaggctttt gagcatccca atgttgtccg
     2281 gctgatggac gtctgtgcca catcccgaac tgaccgggag atcaaggtaa ccctggtgtt
     2341 tgagcatgta gaccaggacc taaggacata tctggacaag gcacccccac caggcttgcc
     2401 agccgaaacg atcaaggatc tgatgcgcca gtttctaaga ggcctagatt tccttcatgc
     2461 caattgcatc gttcaccgag atctgaagcc agagaacatt ctggtgacaa gtggtggaac
     2521 agtcaagctg gctgactttg gcctggccag aatctacagc taccagatgg cacttacacc
     2581 cgtggttgtt acactctggt accgagctcc cgaagttctt ctgcagtcca catatgcaac
     2641 acctgtggac atgtggagtg ttggctgtat ctttgcagag atgtttcgtc gaaagcctct
     2701 cttctgtgga aactctgaag ccgaccagtt gggcaaaatc tttgacctga ttgggctgcc
     2761 tccagaggat gactggcctc gagatgtatc cctgccccgt ggagcctttc cccccagagg
     2821 gccccgccca gtgcagtcgg tggtacctga gatggaggag tcgggagcac agctgctgct
     2881 ggaaatgctg acttttaacc cacacaagcg aatctctgcc tttcgagctc tgcagcactc
     2941 ttatctacat aaggatgaag gtaatccgga gtgagtcgac cctgtggaat gtgtgtcagt
     3001 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3061 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3121 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3181 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3241 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3301 gaggcctagg cttttgcaaa aagcttacca tgaccgagta caagcccacg gtgcgcctcg
     3361 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     3421 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     3481 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     3541 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     3601 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     3661 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     3721 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     3781 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     3841 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     3901 ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg
     3961 aaaggagcgc acgaccccat gcatcgataa aataaaagat tttatttagt ctccagaaaa
     4021 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt
     4081 gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag
     4141 atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     4201 cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca
     4261 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc
     4321 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     4381 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     4441 gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag
     4501 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
     4561 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac
     4621 atgcagcatg tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca
     4681 tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4741 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4801 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4861 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4921 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4981 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5041 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5101 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5161 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5221 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5281 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5341 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5401 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5461 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5521 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5581 atgagattat caaaaaggat cttcacctag atccttttgc ggccgcaaat caatctaaag
     5641 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5701 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5761 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5821 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5881 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5941 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6001 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6061 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6121 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6181 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6241 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6301 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6361 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6421 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6481 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6541 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6601 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccac
//
