LOCUS       sequence-428680-        8173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EGFP-Cyclin F in mammalian cells from a retroviral 
            vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8173)
  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 8173)
  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 F
FEATURES             Location/Qualifiers
     source          1..8173
                     /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"
     promoter        4481..4837
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4503..4523)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4688..4823
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4847..5446
                     /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(4847..4866)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5343..5363
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5667..6136
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6266..6283)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6437..7025)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6517..6536)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7185..7976,7977..8045))
                     /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     7808..7827
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8046..8150)
                     /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 atggggagcg gcggcgtggt ccactgtagg
     2161 tgtgccaagt gtttctgtta tcctacaaag cgaagaataa ggaggaggcc ccgaaacctg
     2221 accatcttga gtctccccga agatgtgctc tttcacatcc tgaaatggct ttctgtagag
     2281 gacatcctgg ccgtccgagc tgtacactcc cagctgaagg acctggtgga caaccacgcc
     2341 agtgtgtggg catgtgccag cttccaggag ctgtggccgt ctccagggaa cctgaagctc
     2401 tttgaaaggg ctgctgaaaa ggggaatttc gaagctgctg tgaagctggg catagcctac
     2461 ctctacaatg aaggcctgtc tgtgtctgat gaggcccgcg cagaagtgaa tggcctgaag
     2521 gcctctcgct tcttcagtct cgctgagcgg ctgaatgtgg gtgccgcacc tttcatctgg
     2581 ctcttcatcc gccctccgtg gtcggtgagc ggaagctgct gcaaggccgt ggttcacgag
     2641 agcctcaggg cagagtgcca gctgcagagg actcacaaag catccatatt gcactgcttg
     2701 ggcagagtgc tgagtctgtt cgaggatgag gagaagcagc agcaggccca tgacctgttt
     2761 gaggaggctg ctcatcaggg atgtctgacc agctcctacc tcctctggga aagcgacagg
     2821 aggacagatg tgtcagatcc tgggcgatgc ctccacagct tccgaaaact cagggactac
     2881 gctgccaaag gctgctggga agcgcagctg tctttagcca aagcctgtgc aaatgcaaac
     2941 cagcttggac tggaggtgag agcttccagt gagatcgtct gccagctatt tcaggcttcc
     3001 caggctgtca gtaaacaaca agtcttctcc gtgcagaagg gactcaatga cacaatgagg
     3061 tacattctga tcgactggct ggtggaagtt gccaccatga aggacttcac aagcctgtgc
     3121 ctgcacctga ccgtggagtg tgtggaccgg tacctgcgga ggaggctggt gccgcggtac
     3181 aggctccagc tgctgggcat cgcctgcatg gtcatctgca cccggtttat cagtaaagag
     3241 atcctgacca tccgggaggc cgtatggctc acggacaaca cttacaagta cgaggacctg
     3301 gtgagaatga tgggcgagat cgtctccgcc ttggaaggga agattcgagt ccccactgtg
     3361 gtggattaca aggaggtcct gctgacgcta gtccctgtgg agctgagaac ccagcacctg
     3421 tgcagcttcc tctgcgagct ctccctgctg cacaccagcc tgtccgccta cgccccagcc
     3481 cgcctggctg ccgcagccct gctcctggcc agactgacgc acgggcagac acagccctgg
     3541 accactcagc tgtgggacct caccggattc tcctatgaag acctcattcc ctgcgtcttg
     3601 agcctccata agaagtgctt ccatgatgac gcccccaagg actacaggca agtctctctg
     3661 accgccgtga agcagcggtt tgaggacaag cgctatggag aaatcagcca ggaagaggtg
     3721 ctgagctaca gccagttgtg tgctgcatta ggagtgacac aagacagccc cgaccccccg
     3781 actttcctca gcacagggga gatccacgcc ttcctcagct ctccctcggg gcggagaacc
     3841 aaacggaagc gggagaacag cctccaggaa gacagaggca gcttcgttac cacccccact
     3901 gcggagctgt ccagccagga ggagacgctg ctgggcagct tcctcgactg gagcctggac
     3961 tgctgctctg gctatgaagg cgaccaggag agtgagggcg agaaggaggg cgacgtgaca
     4021 gctcccagcg gcatcctcga tgtcaccgtg gtctacctga acccagaaca gcattgctgc
     4081 caggaatcca gtgatgagga ggcttgtcca gaggacaagg gaccccagga cccacaggca
     4141 ctggcgctgg acacccagat ccctgcaacc cctggaccca aacccctggt ccgcaccagc
     4201 cgggagccag ggaaggacgt cacgacctca gggtactcct ccgtcagcac cgcaagtccc
     4261 acaagctccg tggacggtgg cttgggggcc ctgccccaac ctacctcagt gctgtccctg
     4321 gacagtgact cgcacacaca gccctgccac catcaggcca ggaagtcatg tttacagtgt
     4381 cgtcccccaa gtcccccgga gagcagtgtt ccccagcaac aggtgaagcg gataaaccta
     4441 tgcatacaca gtgaggagga ggacatgaac ctgggccttg tgaggctgta agtcgaccct
     4501 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4561 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4621 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4681 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4741 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4801 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttaccatga ccgagtacaa
     4861 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     4921 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     4981 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     5041 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     5101 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     5161 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     5221 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     5281 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     5341 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     5401 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc cgccccacga
     5461 cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca tcgataaaat aaaagatttt
     5521 atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc
     5581 ttaagtaacg ccattttgca aggcatggaa aaatacataa ctgagaatag agaagttcag
     5641 atcaaggtca ggaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag
     5701 cagttcctgc cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag
     5761 gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat
     5821 gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga
     5881 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     5941 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag
     6001 tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc
     6061 atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt
     6121 cagcgggggt ctttcacatg cagcatgtat caaaattaat ttggtttttt ttcttaagta
     6181 tttacattaa atggccatag ttgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6241 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6301 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6361 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6421 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6481 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6541 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6601 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6661 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6721 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6781 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6841 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6901 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6961 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7021 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7081 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttgcggc
     7141 cgcaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     7201 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     7261 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     7321 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7381 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7441 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7501 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7561 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7621 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7681 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7741 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7801 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7861 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7921 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7981 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     8041 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     8101 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     8161 cgaaaagtgc cac
//
