LOCUS       sequence-473273-        8316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient overexpression of E2F7 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8316)
  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 8316)
  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 2xFLAG-2xSTREP E2F7
FEATURES             Location/Qualifiers
     source          1..8316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        177..556
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        557..760
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     711..731
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     805..824
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        805..823
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             865..888
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             904..927
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             943..966
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1003..1026
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3849..3866)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3855..4079
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4125..4553
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4212..4231)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4422..4443
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4562..4582)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4567..4896
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4747..4882
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4809..4828
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4963..5757
                     /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(5017..5036)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5627..5646
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5931..6052
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5968..5987)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6022..6041
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6101..6117)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6101..6117)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6114..6136)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6125..6141
                     /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(6149..6155,6156..6173,6174..6179))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6194..6215
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6332..6349)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6503..7091)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6583..6602)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7262..8053,8054..8122))
                     /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     7885..7904
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8123..8227)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(8302..8316,1..5))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
       61 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      121 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      181 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      241 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      301 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      361 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      421 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      481 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      541 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      601 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      661 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      721 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      781 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      841 gcgtttaaac ttaagcttac catggactac aaggacgacg atgacaaggt accaggatct
      901 atggattata aagatgatga tgataaaggg tcggccgcca gctggagcca ccctcagttc
      961 gagaagggag gaggaagcgg cggaggcagc ggaggaggaa gctggagcca cccgcagttc
     1021 gagaaaggag ctggatccag tgtggtggaa ttctgcagat atccagcaca gtggcggccg
     1081 cgcgaggtaa attgtttaac actaaaagac ctgatcagcc ccaggcagcc cagactagat
     1141 tttgcagttg aagatgggga aaatgcacaa aaggaaaata tatttgttga tcgatcaagg
     1201 atggccccga agactccaat aaaaaatgaa ccaattgatt tatcgaagca aaaaaaattt
     1261 actccagaaa gaaatcccat tactccagtt aagcttgttg acagacagca agcggaacca
     1321 tggacaccca cagctaacct gaagatgctc attagtgctg ccagcccaga tataagggac
     1381 cgggagaaga aaaagggact attccgaccc attgaaaaca aggacgatgc atttacagat
     1441 tctctacagc ttgatgttgt tggggacagt gctgtggacg aatttgaaaa gcaaaggcca
     1501 agcagaaaac agaaaagttt aggactcctg tgccagaagt ttctagctcg ctatccaagt
     1561 tatcccttgt caactgagaa aactaccatc tccctagatg aagttgctgt cagtcttggt
     1621 gtggaaagga gacgcatcta tgacattgta aatgtgctgg agtcgctgca tctggtcagc
     1681 cgggtggcta agaatcagta tggctggcat ggacggcaca gcctgccaaa aaccctgagg
     1741 aacctccaga gactaggaga ggagcagaaa tatgaagagc aaatggccta cctccaacag
     1801 aaagagctgg acctgataga ttataaattt ggagaacgta aaaaagatgg tgatccagat
     1861 tcccaggaac aacagttact ggatttctct gaacccgact gtccctcttc atctgcaaac
     1921 agtagaaaag acaagtctct gagaattatg agccagaagt ttgtcatgct gttcctcgtc
     1981 tccaaaacca agattgtcac tctggatgtg gctgccaaaa tactgataga agaaagccaa
     2041 gatgccccag accatagtaa atttaaaaca aaggtacgac gcctctatga catagccaat
     2101 gttctgacca gcttggctct gataaagaaa gtgcatgtaa cagaagagcg aggtcgtaaa
     2161 ccagccttca agtggatcgg gcctgtggac ttcagctcaa gtgatgaaga actggtggat
     2221 gtttctgcat ctgtcttacc agaattgaaa agagaaacat atggccagat tcaagtctgt
     2281 gcaaaacaga agctggctcg ccatggttct tttaacacag ttcaggcttc tgagaggatc
     2341 cagaggaaag tgaactcaga accgagcagc ccgtacagag aagaacaagg atcaggtggc
     2401 tactctttag aaattggaag cctggcagct gtctatagac agaaaataga agacaattca
     2461 cagggaaaag cctttgccag taagagagtg gtgcctccat caagcagctt ggaccctgtt
     2521 gctcctttcc ctgtcctctc tgttgaccca gaatattgtg ttaatccttt agcccaccca
     2581 gtattttctg ttgctcagac ggacctgcag gcattctcca tgcagaacgg tctgaatgga
     2641 caagtggatg tctcacttgc ttctgcagcc tctgctgtgg agagcctgaa gccagcactc
     2701 cttgctggcc agcctctagt gtatgtgccc tctgcctcac tgttcatgct gtatggaagt
     2761 ctgcaggagg gaccagcgtc agggtcaggg tcagagaggg atgacagaag ctcagaagcc
     2821 ccagccacag tagagctgtc atctgcaccc tcagctcaga agcgcctctg tgaggagagg
     2881 aaacctcagg aggaggatga gccagccact aaaaggcaaa gtagggaata tgaagacggc
     2941 ccgctgtcgc ttgtcatgcc caagaaaccc tcagattcca cagaccttgc ctctcccaag
     3001 actatgggta acagggcatc tatacccctc aaagacattc atgtgaatgg ccaactccct
     3061 gctgcagaag agatttcagg aaaggcaaca gcaaactctc ttgtttcttc tgagtgggga
     3121 aatccttcaa gaaatacaga tgttgaaaag ccttcaaaag aaaatgaaag caccaaagag
     3181 ccttctttgc tacaatatct ttgtgtgcag tctcctgcag gattaaatgg tttcaatgta
     3241 cttttatctg gcagtcaaac cccccctact gtgggcccgt cctcaggtca gctgccgtct
     3301 ttcagtgtcc cttgcatggt cttaccatct ccacctctgg gcccttttcc tgttctctat
     3361 tctcctgcaa tgccgggccc ggtttcttct actcttggtg ctctcccaaa cacaggacct
     3421 gtgaatttca gcttgcctgg ccttggatca atagcccagc ttctcgtcgg ccccacagct
     3481 gtggttaatc caaagtcgtc cacactccct tctgcagacc ctcagcttca gagtcagccc
     3541 tcactaaacc taagtccagt gatgtcaagg tcacacagtg tcgtccaaca acctgagtcc
     3601 cccgtttacg tgggacatcc agtctcagta gtaaaattac atcagtcacc agttccagtg
     3661 acccccaaga gcatccaacg cacacatcgt gagacgtttt tcaagacacc cggcagcctt
     3721 ggagaccctg tcctgaagag aagagaaagg aaccagtcac gaaacaccag ctcggcccag
     3781 aggagactag aaatccccag cggcggcgct gactaagtct agagggcccg tttaaacccg
     3841 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3901 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3961 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     4021 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     4081 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4141 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4201 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4261 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4321 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4381 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4441 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4501 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4561 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4621 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4681 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4741 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4801 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4861 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4921 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4981 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5041 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5101 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     5161 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5221 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5281 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5341 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5401 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5461 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5521 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5581 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5641 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5701 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5761 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5821 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5881 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5941 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     6001 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     6061 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     6121 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6181 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6241 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6301 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6361 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6421 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6481 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6541 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6601 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6661 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6721 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6781 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6841 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6901 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6961 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7021 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     7081 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     7141 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     7201 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7261 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7321 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7381 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7441 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7501 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7561 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7621 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7681 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7741 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7801 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7861 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7921 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7981 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     8041 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     8101 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     8161 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     8221 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     8281 tcccctatgg tgcactctca gtacaatctg ctctga
//
