LOCUS       sequence-473712-        6573 bp    DNA     circular SYN 22-MAY-2025
DEFINITION  stable expression of E2F1 using virus infection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6573)
  AUTHORS   Clijsters L, Hoencamp C, Calis JJA, Marzio A, Handgraaf SM, Cuitino 
            MC, Rosenberg BR, Leone G, Pagano M
  TITLE     Cyclin F Controls Cell-Cycle Transcriptional Outputs by Directing 
            the Degradation of the Three Activator E2Fs.
  JOURNAL   Mol Cell. 2019 Jun 20;74(6):1264-1277.e7. doi: 
            10.1016/j.molcel.2019.04.010. Epub 2019 May 23.
  PUBMED    31130363
REFERENCE   2  (bases 1 to 6573)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.molcel.2019.04.010"; journalName: "Mol Cell"; date: 
            "2019-06-20- 20"; volume: "74"; issue: "6"; pages: "1264-1277.e7"
COMMENT     Sequence Label: pBABE-puro-2xFLAG-2xSTREP-E2F1
FEATURES             Location/Qualifiers
     source          1..6573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    40..239
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             304..773
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        804..908
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(909..977,978..1769)
                     /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     complement(1127..1146)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1929..2517
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2418..2437
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2671..2688
                     /label=L4440
                     /note="L4440 vector, forward primer"
     LTR             2818..3287
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(3508..4107)
                     /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(3591..3611)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4088..4107
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(4117..4446)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4131..4266
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4431..4451
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(5823..5846)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(5883..5906)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(5922..5945)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(5961..5984)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(5996..6412)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(6012..6028)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(6044..6066)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
ORIGIN
        1 catatcctga cggggtcgga ttccacatcg actcccttcc tcaggtcggg ccacaaaaac
       61 ggcccccaaa gtccctggga cgtctcccag ggttgcggcc gggtgttcag aactcgtcag
      121 ttccaccacg ggtccgccag atacagagct agttagctaa ctagtaccga cgcaggcgca
      181 taaaatcagt catagacact agacaatcgg acagacacag ataagttgct ggccagcttg
      241 cctcccggtg gtgggtcggt ggtccctggg caggggtctc ccgatcccgg acgagccccc
      301 aaatgaaaga cccccgctga cgggtagtca atcactcaga ggagaccctc ccaaggaaca
      361 gcgagaccac aagtcggatg caactgcaag agggtttatt ggatacacgg gtacccgggc
      421 gactcagtca atcggaggac tggcgccccg agtgaggggt tgtgggctct tttattgagc
      481 tcggggagca gaagcgcgcg aacagaagcg agaagcgaac tgattggtta gttcaaataa
      541 ggcacagggt catttcaggt ccttggggca ccctggaaac atctgatggt tctctagaaa
      601 ctgctgaggg ctggaccgca tctggggacc atctgttctt ggccctgagc cggggcagga
      661 actgcttacc acagatatcc tgtttggccc atattcagct gttccatctg ttcttggccc
      721 tgagccgggg caggaactgc ttaccacaga tatcctgttt ggcccatatt caggctgcag
      781 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
      841 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      901 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      961 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     1021 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     1081 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     1141 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     1201 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     1261 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     1321 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     1381 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     1441 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     1501 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     1561 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     1621 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     1681 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     1741 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1801 agattgattt gcggccgcaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     1861 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1921 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1981 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2041 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     2101 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     2161 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     2221 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     2281 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2341 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2401 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2461 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2521 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2581 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     2641 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     2701 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg caactatggc
     2761 catttaatgt aaatacttaa gaaaaaaaac caaattaatt ttgatacatg ctgcatgtga
     2821 aagacccccg ctgacgggta gtcaatcact cagaggagac cctcccaagg aacagcgaga
     2881 ccacaagtcg gatgcaactg caagagggtt tattggatac acgggtaccc gggcgactca
     2941 gtcaatcgga ggactggcgc cccgagtgag gggttgtggg ctcttttatt gagctcgggg
     3001 agcagaagcg cgcgaacaga agcgagaagc gaactgattg gttagttcaa ataaggcaca
     3061 gggtcatttc aggtccttgg ggcaccctgg aaacatctga tggttctcta gaaactgctg
     3121 agggctggac cgcatctggg gaccatctgt tcttggccct gagccggggc aggaactgct
     3181 taccacagat atcctgtttg gcccatattc agctgttcca tctgttcttg gccctgagcc
     3241 ggggcaggaa ctgcttacca cagatatcct gtttggccca tattcagctg ttccatctgt
     3301 tcctgacctt gatctgaact tctctattct cagttatgta tttttccatg ccttgcaaaa
     3361 tggcgttact taagctagct tgccaaacct acaggtgggg tctttcattc cccccttttt
     3421 ctggagacta aataaaatct tttattttat cgatgcatgg ggtcgtgcgc tcctttcggt
     3481 cgggcgctgc gggtcgtggg gcgggcgtca ggcaccgggc ttgcgggtca tgcaccaggt
     3541 gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc gctcgtagaa
     3601 ggggaggttg cggggcgcgg aggtctccag gaaggcgggc accccggcgc gctcggccgc
     3661 ctccactccg gggagcacga cggcgctgcc cagacccttg ccctggtggt cgggcgagac
     3721 gccgacggtg gccaggaacc acgcgggctc cttgggccgg tgcggcgcca ggaggccttc
     3781 catctgttgc tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg ggccgatctc
     3841 ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc cagaccgcca ccgcggcgcc
     3901 gtcgtccgcg acccacacct tgccgatgtc gagcccgacg cgcgtgagga agagttcttg
     3961 cagctcggtg acccgctcga tgtggcggtc cggatcgacg gtgtggcgcg tggcggggta
     4021 gtcggcgaac gcggcggcga gggtgcgtac ggccctgggg acgtcgtcgc gggtggcgag
     4081 gcgcaccgtg ggcttgtact cggtcatggt aagctttttg caaaagccta ggcctccaaa
     4141 aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg cctctgcata
     4201 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     4261 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     4321 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     4381 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     4441 acacacattc cacagggtcg accactgtgc tggcgaattc ctacgtacca ccacactggt
     4501 cagaaatcca ggggggtgag gtccccaaag tcacagtcga agaggtctct gatgccctcg
     4561 ccctcctcga ggccgaagtg gtagtcgagg gcctcgtggg gtggggaaag gctgatgaac
     4621 tcctcaggga ggaggccgga gaagtcctcc cgcacatgct ccaggagcga gtcggccgcc
     4681 accagcgggg acaggcggtc ctcgtccacg ggagcccgca ggctgcccat ccgggacaac
     4741 agcggttttt gctccaggct gagtagagac tggctgggat ctgtggtgag ggatgagggg
     4801 ggagatgatg gtggtggtga cactatggtg gcagagtcag tggccctgtt ctcctcctca
     4861 gaagtgacct cctgggatgg ggtcttccca gggctgatcc cacctacggt ctcctcaggg
     4921 cacaggaaaa catcgatcgg gccttgtttg ctcttaaggg agatctgaaa gttctccgaa
     4981 gagtccacgg cttggagctg ggtctcagga ggggctttga tcaccataac catctgctct
     5041 gcagggtctg caatgctacg aaggtcctga cacgtcacgt aggccaggcg ctggctgtca
     5101 gtgtcctcgg agagcaggcg cagctgcgta gtacagatat tcatcaggtg gtccagctgc
     5161 tgctcgctct cctgcagctg tcggaggtcc tgggtcaacc cctcaagccg tccgccgacg
     5221 cccactgtgg tgtggctgcc cagccactgg atgtggttct tggacttctt ggcaatgagc
     5281 tggatgccct caaggacgtt ggtgatgtca tagatgcgcc gcttctgcac cttcagcacc
     5341 tcggcagccc agttcaggtc gacgacaccg tcagccgagt ggctcagcag ctccaggaag
     5401 cgcttggtgg tcagattcag tgaggtctca tagcgtgact tctcccccgg ggatttcaca
     5461 ccttttcctg gatggcggcc tctgccccga gctggcccac tgctctcggc caggtactga
     5521 tggtcagttt ccaggtccag cctccgcttc accggcgggc ggccgagcgc gggccgcggc
     5581 gcactgggtg tgggccgggg cgcctgcggt gtggcgaaga gcagcaggtc agggtcgcag
     5641 gggccggcgg cgggcgccgc ggggccggtg ggagccggcg gggcgctggc gtcctgcgcg
     5701 gcggagatga tgacgatctg cgaggagtcg agcagccgca gcgcgccggc cccgagcagg
     5761 gcctccagcg ccggcgcgca tgggccgccc gcaggggccc cggccaaggc ggatccagct
     5821 cctttctcga actgcgggtg gctccagctt cctcctccgc tgcctccgcc gcttcctcct
     5881 cccttctcga actgagggtg gctccagctg gcggccgacc ctttatcatc atcatcttta
     5941 taatccatag atcctggtac cttgtcatcg tcgtccttgt agtccatggt cggccggcgc
     6001 ctagagaagg agtgagggct ggataaaggg aggatcgagg cggggtcgaa cgaggaggtt
     6061 caagggggag agacggggcg gatggaggaa gaggaggcgg aggcttaggg tgtacaaagg
     6121 gcttgaccca gggagggggg tcaaaagcca aggcttccca ggtcacgatg taggggacct
     6181 ggtctgggtg tccatgcggg ccaggtgaaa agaccttgat cttaacctgg gtgatgaggt
     6241 ctcggttaaa ggtgccgtct cgcggccatc cgacgttaaa ggttggccat tctgcagagc
     6301 agaaggtaac ccaacgtctc ttcttgacat ctaccgactg gttgtgagcg atccgctcga
     6361 catctttcca gtgacctaag gtcaaactta agggagtggt aacagtctgg ccctaatttt
     6421 cagacaaata cagaaacaca gtcagacaga gacaacacag aacgatgctg cagcgctgca
     6481 gcagacaaga cgcgcggctt cggttccaaa ccgaaagcaa aaattcagac ggaggcggga
     6541 actgttttag gttctcgtct cctaccagaa cca
//
