LOCUS       sequence-473633-        6793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient overexpression of E2F1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6793)
  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 6793)
  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 E2F1
FEATURES             Location/Qualifiers
     source          1..6793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      303..891
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     792..811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1045..1062
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1179..1200
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1215..1220,1221..1238,1239..1245)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1253..1269
                     /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)."
     primer_bind     1258..1280
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1277..1293
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1277..1293
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1342..1463
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1353..1372)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1407..1426
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1637..2431)
                     /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(1748..1767)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2358..2377
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2498..2827)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2512..2647
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2566..2585)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2812..2832
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2841..3269)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2951..2972)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3163..3182
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3315..3539
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3528..3545
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        3565..3583
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3565..3582
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(4993..5019)
                     /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"
     CDS             complement(4999..5025)
                     /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             complement(4999..5025)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5047..5066)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5048..5066)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5111..5314)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5140..5160)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5315..5694
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5866..5885
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5960..6064
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6065..6133,6134..6793,1..132)
                     /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(6283..6302)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
       61 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
      121 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
      181 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
      241 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
      301 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
      361 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      421 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
      481 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      541 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      601 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      661 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      721 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      781 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      841 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      901 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      961 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     1021 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     1081 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     1141 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     1201 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     1261 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag
     1321 gtcgacggta tacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     1381 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1441 ttataagctg caataaacaa gttggggtgg gcgaagaact ccagcatgag atccccgcgc
     1501 tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag
     1561 gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg
     1621 aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     1681 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     1741 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     1801 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     1861 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg agcctggcga
     1921 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     1981 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     2041 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     2101 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     2161 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     2221 ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg
     2281 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     2341 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     2401 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     2461 cagatccgaa aatggatata caagctcccg ggagcttttt gcaaaagcct aggcctccaa
     2521 aaaagcctcc tcactacttc tggaatagct cagaggcaga ggcggcctcg gcctctgcat
     2581 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
     2641 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt
     2701 gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga
     2761 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact
     2821 gacacacatt ccacagaatt aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2881 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     2941 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3001 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3061 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3121 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3181 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     3241 cgcgcttaat gcgccgctac agggcgcgtg gggatacccc ctagagcccc agctggttct
     3301 ttccgcctca gaagccatag agcccaccgc atccccagca tgcctgctat tgtcttccca
     3361 atcctccccc ttgctgtcct gccccacccc accccccaga atagaatgac acctactcag
     3421 acaatgcgat gcaatttcct cattttatta ggaaaggaca gtgggagtgg caccttccag
     3481 ggtcaaggaa ggcacggggg aggggcaaac aacagatggc tggcaactag aaggcacagt
     3541 cgaggctgat cagcgagctc tagcatttag gtgacactat agaatagggc cctctagatg
     3601 catgctcgag cggccgccag tgtgatggat atctgcagaa tttcagaaat ccaggggggt
     3661 gaggtcccca aagtcacagt cgaagaggtc tctgatgccc tcgccctcct cgaggccgaa
     3721 gtggtagtcg agggcctcgt ggggtgggga aaggctgatg aactcctcag ggaggaggcc
     3781 ggagaagtcc tcccgcacat gctccaggag cgagtcggcc gccaccagcg gggacaggcg
     3841 gtcctcgtcc acgggagccc gcaggctgcc catccgggac aacagcggtt tttgctccag
     3901 gctgagtaga gactggctgg gatctgtggt gagggatgag gggggagatg atggtggtgg
     3961 tgacactatg gtggcagagt cagtggccct gttctcctcc tcagaagtga cctcctggga
     4021 tggggtcttc ccagggctga tcccacctac ggtctcctca gggcacagga aaacatcgat
     4081 cgggccttgt ttgctcttaa gggagatctg aaagttctcc gaagagtcca cggcttggag
     4141 ctgggtctca ggaggggctt tgatcaccat aaccatctgc tctgcagggt ctgcaatgct
     4201 acgaaggtcc tgacacgtca cgtaggccag gcgctggctg tcagtgtcct cggagagcag
     4261 gcgcagctgc gtagtacaga tattcatcag gtggtccagc tgctgctcgc tctcctgcag
     4321 ctgtcggagg tcctgggtca acccctcaag ccgtccgccg acgcccactg tggtgtggct
     4381 gcccagccac tggatgtggt tcttggactt cttggcaatg agctggatgc cctcaaggac
     4441 gttggtgatg tcatagatgc gccgcttctg caccttcagc acctcggcag cccagttcag
     4501 gtcgacgaca ccgtcagccg agtggctcag cagctccagg aagcgcttgg tggtcagatt
     4561 cagtgaggtc tcatagcgtg acttctcccc cggggatttc acaccttttc ctggatggcg
     4621 gcctctgccc cgagctggcc cactgctctc ggccaggtac tgatggtcag tttccaggtc
     4681 cagcctccgc ttcaccggcg ggcggccgag cgcgggccgc ggcgcactgg gtgtgggccg
     4741 gggcgcctgc ggtgtggcga agagcagcag gtcagggtcg caggggccgg cggcgggcgc
     4801 cgcggggccg gtgggagccg gcggggcgct ggcgtcctgc gcggcggaga tgatgacgat
     4861 ctgcgaggag tcgagcagcc gcagcgcgcc ggccccgagc agggcctcca gcgccggcgc
     4921 gcatgggccg cccgcagggg ccccggccaa ggccagaatt ccagcacact ggcggccgtt
     4981 actagtggat ccgagggagg cgtagtcggg cacgtcgtag gggtaggcca tggtaagctt
     5041 gggtctccct atagtgagtc gtattaattt cgataagcca gtaagcagtg ggttctctag
     5101 ttagccagag agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg
     5161 tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca
     5221 aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc
     5281 cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt
     5341 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg
     5401 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta
     5461 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
     5521 ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg
     5581 gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat
     5641 gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa tgtcaacgcg
     5701 tatatctggc ccgtacatcg cgaagcagcg caaaacgcct aaccctaagc agattcttca
     5761 tgcaattgtc ggtcaagcct tgccttgttg tagcttaaat tttgctcgcg cactactcag
     5821 cgacctccaa cacacaagca gggagcagat actggcttaa ctatgcggca tcagagcaga
     5881 ttgtactgag agtgcaccat aggggatcgg gagatctccc gatccgtcga cgtcaggtgg
     5941 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     6001 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     6061 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     6121 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     6181 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     6241 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     6301 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     6361 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6421 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6481 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6541 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6601 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     6661 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     6721 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6781 gtctcgcggt atc
//
