LOCUS       sequence-473628-        6877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient overexpression of E2F3A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6877)
  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 6877)
  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: 
            "10.1016/j.molcel.2019.04.010"; journalName: "Mol Cell"; date: 
            "2019-06-20- 20"; volume: "74"; issue: "6"; pages: "1264-1277.e7"
COMMENT     Sequence Label: pcDNA3 HA E2F3A
FEATURES             Location/Qualifiers
     source          1..6877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(72..88)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(72..88)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(85..107)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    96..112
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(120..126,127..144,145..150))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    165..186
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(303..320)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(474..1062)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(554..573)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1233..2024,2025..2093))
                     /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     1856..1875
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2094..2198)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2273..2292)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2464..2843
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2844..3047
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2998..3018
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3092..3111
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3092..3110
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3133..3159
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             3133..3159
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3139..3165
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     promoter        complement(4659..4677)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4660..4677)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4697..4714)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4703..4927
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4973..5401
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5060..5079)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5270..5291
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5410..5430)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5415..5744
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5595..5730
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5657..5676
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5811..6605
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5865..5884)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6475..6494
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    join(6779..6877,1..23)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6816..6835)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     join(6870..6877,1..12)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
       61 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
      121 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
      181 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
      241 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
      301 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      361 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      421 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      481 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      541 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      601 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      661 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      721 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      781 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      841 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      901 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      961 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1021 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1081 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     1141 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     1201 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1261 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1321 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1381 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1441 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1501 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1561 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1621 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1681 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1741 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1801 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1861 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1921 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1981 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     2041 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     2101 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     2161 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     2221 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     2281 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     2341 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     2401 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     2461 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     2521 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     2581 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     2641 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     2701 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     2761 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     2821 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     2881 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     2941 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     3001 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
     3061 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
     3121 cttaccatgg cctaccccta cgacgtgccc gactacgcct ccctcggatc cactagtaac
     3181 ggccgccagt gtgctggaat tctgagaaag ggaatccagc ccgctctgga gcagtacctg
     3241 gtgaccgccg ggggtgggga gggggcggct gtcgtcgccg ccgccgctgc agcctccatg
     3301 gacaaaaggg cactgctagc cagccccggc ttcgccgccg ccgccgccgc tgccgccgcc
     3361 ccgggcgcgt acatccagat cctcaccacg aacacttcca ccacctcctg ttcctcctcc
     3421 ctccaaagcg gcgccgtagc cgccggcccc ctcctcccca gtgcccccgg cgcggagcag
     3481 accgccggca gcctcctcta caccacgccg cacggaccct ccagcagagc cgggctgctg
     3541 cagcagccac cagcgctggg acgcggcggc agcggcggcg gcggcggccc tccggcaaag
     3601 cgaaggctgg agctaggaga aagcggtcat cagtacctct cagatggttt aaaaaccccc
     3661 aagggcaaag gaagagctgc actacgaagt ccagatagtc caaaaactcc aaaatctccc
     3721 tcagaaaaaa cgcggtatga tacgtctctt ggtctgctca ccaagaagtt cattcagctc
     3781 ctgagccagt cacccgatgg ggtattggat ttgaacaagg cagcagaagt gctaaaagtg
     3841 caaaagagaa ggatttatga tatcaccaac gttctggaag gcatccacct cattaagaag
     3901 aagtctaaaa acaacgtcca atggatgggc tgcagtctgt ctgaggatgg gggcatgctg
     3961 gcccagtgtc aaggcctgtc aaaagaagtg accgagctca gtcaggaaga gaagaaatta
     4021 gatgaactga tccaaagctg caccctggac ctcaaactgt taaccgagga ttcagagaat
     4081 caaaggttag cttatgttac atatcaagat attcgaaaaa ttagtggcct taaagaccaa
     4141 actgttatag ttgtgaaagc ccctccagaa acaagacttg aagtgcctga ctcaatagag
     4201 agcctacaaa tacatttggc aagtacccaa gggcccattg aggtttactt atgtccagaa
     4261 gagactgaaa cacacagtcc aatgaaaaca aacaaccaag accacaatgg gaatatccct
     4321 aaacccgctt ccaaagactt ggcttcaacc aactcaggac atagcgattg ctcagtttct
     4381 atgggaaacc tttctcctct ggcctcccca gccaacctct tacagcagac tgaggaccaa
     4441 attccttcca acctagaagg accgtttgtg aacttactgc ctcccctgct gcaagaggac
     4501 tatctcctga gcctcgggga ggaggaaggc atcagcgatc tcttcgatgc ttacgatttg
     4561 gaaaagctcc cactggtgga agacttcatg tgtagttgaa attctgcaga tatccatcac
     4621 actggcggcc gctcgagcat gcatctagag ggccctattc tatagtgtca cctaaatgct
     4681 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     4741 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     4801 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     4861 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     4921 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     4981 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     5041 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     5101 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     5161 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     5221 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5281 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     5341 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5401 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     5461 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     5521 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     5581 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5641 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     5701 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     5761 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     5821 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     5881 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     5941 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     6001 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     6061 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     6121 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     6181 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     6241 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     6301 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     6361 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     6421 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     6481 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     6541 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     6601 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     6661 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     6721 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     6781 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6841 taaagcattt ttttcactgc attctagttg tggtttg
//
