LOCUS       sequence-436482-        8419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EGFP-Cyclin A2 in mammalian cells from a retroviral 
            vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8419)
  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 8419)
  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: EGFP-Cyclin A2
FEATURES             Location/Qualifiers
     source          1..8419
                     /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      1393..1402
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1399..1401,1402..1404,1405..2115)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1399..1401,1402..1404,1405..2115)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1444..1465)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1705..1724)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2052..2073
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4749..4769)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4754..5083
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4934..5069
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5093..5692
                     /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(5093..5112)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5589..5609
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5913..6382
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6512..6529)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6683..7271)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6763..6782)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7431..8222,8223..8291))
                     /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     8054..8073
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8292..8396)
                     /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 ccggatccca gtgtggtggt
     1381 acgctaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     1441 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     1501 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     1561 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     1621 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     1681 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     1741 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     1801 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     1861 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     1921 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1981 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     2041 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     2101 gacgagctgt acaagtccgg actcagatct ttgggcaact ctgcgccggg gcctgcgacc
     2161 cgcgaggcgg gctcggcgct gctagcattg cagcagacgg cgctccaaga ggaccaggag
     2221 aatatcaacc cggaaaaggc agcgcccgtc caacaaccgc ggacccgggc cgcgctggcg
     2281 gtactgaagt ccgggaaccc gcggggtcta gcgcagcagc agaggccgaa gacgagacgg
     2341 gttgcacccc ttaaggatct tcctgtaaat gatgagcatg tcaccgttcc tccttggaaa
     2401 gcaaacagta aacagcctgc gttcaccatt catgtggatg aagcagaaaa agaagctcag
     2461 aagaagccag ctgaatctca aaaaatagag cgtgaagatg ccctggcttt taattcagcc
     2521 attagtttac ctggacccag aaaaccattg gtccctcttg attatccaat ggatggtagt
     2581 tttgagtcac cacatactat ggacatgtca attgtattag aagatgaaaa gccagtgagt
     2641 gttaatgaag taccagacta ccatgaggat attcacacat accttaggga aatggaggtt
     2701 aaatgtaaac ctaaagtggg ttacatgaag aaacagccag acatcactaa cagtatgaga
     2761 gctatcctcg tggactggtt agttgaagta ggagaagaat ataaactaca gaatgagacc
     2821 ctgcatttgg ctgtgaacta cattgatagg ttcctgtctt ccatgtcagt gctgagagga
     2881 aaacttcagc ttgtgggcac tgctgctatg ctgttagcct caaggtttga agaaatatac
     2941 cccccagaag tagcagagtt tgtgtacatt acagatgata cctacaccaa gaaacaagtt
     3001 ctgagaatgg agcatctagt tttgaaagtc cttacttttg acttagctgc tccaacagta
     3061 aatcagtttc ttacccaata ctttctgcat cagcagcctg caaactgcaa agttgaaagt
     3121 ttagcaatgt ttttgggaga attaagtttg atagatgctg acccatacct caagtatttg
     3181 ccatcagtta ttgctggagc tgcctttcat ttagcactct acacagtcac gggacaaagc
     3241 tggcctgaat cattaatacg aaagactgga tataccctgg aaagtcttaa gccttgtctc
     3301 atggaccttc accagaccta cctcaaagca ccacagcatg cacaacagtc aataagagaa
     3361 aagtacaaaa attcaaagta tcatggtgtt tctctcctca acccaccaga gacactaaat
     3421 ctgtaagtcg actataagat ctttgggcaa ctctgcgccg gggcctgcga cccgcgaggc
     3481 gggctcggcg ctgctagcat tgcagcagac ggcgctccaa gaggaccagg agaatatcaa
     3541 cccggaaaag gcagcgcccg tccaacaacc gcggacccgg gccgcgctgg cggtactgaa
     3601 gtccgggaac ccgcggggtc tagcgcagca gcagaggccg aagacgagac gggttgcacc
     3661 ccttaaggat cttcctgtaa atgatgagca tgtcaccgtt cctccttgga aagcaaacag
     3721 taaacagcct gcgttcacca ttcatgtgga tgaagcagaa aaagaagctc agaagaagcc
     3781 agctgaatct caaaaaatag agcgtgaaga tgccctggct tttaattcag ccattagttt
     3841 acctggaccc agaaaaccat tggtccctct tgattatcca atggatggta gttttgagtc
     3901 accacatact atggacatgt caattgtatt agaagatgaa aagccagtga gtgttaatga
     3961 agtaccagac taccatgagg atattcacac ataccttagg gaaatggagg ttaaatgtaa
     4021 acctaaagtg ggttacatga agaaacagcc agacatcact aacagtatga gagctatcct
     4081 cgtggactgg ttagttgaag taggagaaga atataaacta cagaatgaga ccctgcattt
     4141 ggctgtgaac tacattgata ggttcctgtc ttccatgtca gtgctgagag gaaaacttca
     4201 gcttgtgggc actgctgcta tgctgttagc ctcaaggttt gaagaaatat accccccaga
     4261 agtagcagag tttgtgtaca ttacagatga tacctacacc aagaaacaag ttctgagaat
     4321 ggagcatcta gttttgaaag tccttacttt tgacttagct gctccaacag taaatcagtt
     4381 tcttacccaa tactttctgc atcagcagcc tgcaaactgc aaagttgaaa gtttagcaat
     4441 gtttttggga gaattaagtt tgatagatgc tgacccatac ctcaagtatt tgccatcagt
     4501 tattgctgga gctgcctttc atttagcact ctacacagtc acgggacaaa gctggcctga
     4561 atcattaata cgaaagactg gatataccct ggaaagtctt aagccttgtc tcatggacct
     4621 tcaccagacc tacctcaaag caccacagca tgcacaacag tcaataagag aaaagtacaa
     4681 aaattcaaag tatcatggtg tttctctcct caacccacca gagacactaa atcgtaagtc
     4741 gaccctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4801 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4861 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4921 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4981 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     5041 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta ccatgaccga
     5101 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     5161 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     5221 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     5281 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     5341 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     5401 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     5461 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     5521 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     5581 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     5641 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc
     5701 ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcga taaaataaaa
     5761 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     5821 gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa
     5881 gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt
     5941 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc
     6001 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     6061 ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     6121 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     6181 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg
     6241 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
     6301 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     6361 acccgtcagc gggggtcttt cacatgcagc atgtatcaaa attaatttgg ttttttttct
     6421 taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc gcggggagag
     6481 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6541 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6601 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6661 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6721 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6781 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6841 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6901 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6961 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7021 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7081 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     7141 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7201 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7261 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7321 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7381 tgcggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7441 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7501 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7561 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7621 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7681 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7741 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7801 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7861 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7921 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7981 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8041 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8101 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8161 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8221 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8281 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8341 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8401 tttccccgaa aagtgccac
//
