LOCUS       sequence-473574-        7923 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient overexpression of Cyclin F in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7923)
  AUTHORS   D'Angiolella V, Donato V, Forrester FM, Jeong YT, Pellacani C, Kudo 
            Y, Saraf A, Florens L, Washburn MP, Pagano M
  TITLE     Cyclin F-mediated degradation of ribonucleotide reductase M2 
            controls genome integrity and DNA repair.
  JOURNAL   Cell. 2012 May 25;149(5):1023-34. doi: 10.1016/j.cell.2012.03.043.
  PUBMED    22632967
REFERENCE   2  (bases 1 to 7923)
  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.cell.2012.03"; journalName: "Cell"; date: "2012-05-25- 
            25"; volume: "149"; issue: "5"; pages: "1023-34"
COMMENT     Sequence Label: pcDNA3 2xSTREP Cyclin F(LP35/36AA)
FEATURES             Location/Qualifiers
     source          1..7923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(408..1199,1200..1268))
                     /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     1031..1050
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1269..1373)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1448..1467)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1639..2018
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2019..2222
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2173..2193
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2267..2286
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2267..2285
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2327..2350
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2366..2389
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2405..2428
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2465..2488
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(4918..4935)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4924..5148
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5194..5622
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5281..5300)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5491..5512
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5631..5651)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5636..5965
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5816..5951
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5878..5897
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6032..6826
                     /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(6086..6105)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6696..6715
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7000..7121
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7037..7056)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7091..7110
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7170..7186)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7170..7186)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7183..7205)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7194..7210
                     /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(7218..7224,7225..7242,7243..7248))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7263..7284
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7401..7418)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(7572..7923,1..237))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7652..7671)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
       61 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
      121 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      181 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
      241 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
      301 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
      361 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
      421 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
      481 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
      541 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
      601 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
      661 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
      721 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
      781 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
      841 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
      901 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
      961 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1021 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     1081 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1141 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1201 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1261 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1321 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     1381 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     1441 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     1501 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     1561 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     1621 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     1681 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     1741 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     1801 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     1861 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     1921 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     1981 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     2041 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     2101 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     2161 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     2221 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
     2281 atagggagac ccaagctggc tagcgtttaa acttaagctt accatggact acaaggacga
     2341 cgatgacaag gtaccaggat ctatggatta taaagatgat gatgataaag ggtcggccgc
     2401 cagctggagc caccctcagt tcgagaaggg aggaggaagc ggcggaggca gcggaggagg
     2461 aagctggagc cacccgcagt tcgagaaagg agctggatcc agtgtggtgg aattcgggag
     2521 cggcggcgtg gtccactgta ggtgtgccaa gtgtttctgt tatcctacaa agcgaagaat
     2581 aaggaggagg ccccgaaacc tgaccatctt gagtgccgcc gaagatgtgc tctttcacat
     2641 cctgaaatgg ctttctgtag aggacatcct ggccgtccga gctgtacact cccagctgaa
     2701 ggacctggtg gacaaccacg ccagtgtgtg ggcatgtgcc agcttccagg agctgtggcc
     2761 gtctccaggg aacctgaagc tctttgaaag ggctgctgaa aaggggaatt tcgaagctgc
     2821 tgtgaagctg ggcatagcct acctctacaa tgaaggcctg tctgtgtctg atgaggcccg
     2881 cgcagaagtg aatggcctga aggcctctcg cttcttcagt ctcgctgagc ggctgaatgt
     2941 gggtgccgca cctttcatct ggctcttcat ccgccctccg tggtcggtga gcggaagctg
     3001 ctgcaaggcc gtggttcacg agagcctcag ggcagagtgc cagctgcaga ggactcacaa
     3061 agcatccata ttgcactgct tgggcagagt gctgagtctg ttcgaggatg aggagaagca
     3121 gcagcaggcc catgacctgt ttgaggaggc tgctcatcag ggatgtctga ccagctccta
     3181 cctcctctgg gaaagcgaca ggaggacaga tgtgtcagat cctgggcgat gcctccacag
     3241 cttccgaaaa ctcagggact acgctgccaa aggctgctgg gaagcgcagc tgtctttagc
     3301 caaagcctgt gcaaatgcaa accagcttgg actggaggtg agagcttcca gtgagatcgt
     3361 ctgccagcta tttcaggctt cccaggctgt cagtaaacaa caagtcttct ccgtgcagaa
     3421 gggactcaat gacacaatga ggtacattct gatcgactgg ctggtggaag ttgccaccat
     3481 gaaggacttc acaagcctgt gcctgcacct gaccgtggag tgtgtggacc ggtacctgcg
     3541 gaggaggctg gtgccgcggt acaggctcca gctgctgggc atcgcctgca tggtcatctg
     3601 cacccggttt atcagtaaag agatcctgac catccgggag gccgtatggc tcacggacaa
     3661 cacttacaag tacgaggacc tggtgagaat gatgggcgag atcgtctccg ccttggaagg
     3721 gaagattcga gtccccactg tggtggatta caaggaggtc ctgctgacgc tagtccctgt
     3781 ggagctgaga acccagcacc tgtgcagctt cctctgcgag ctctccctgc tgcacaccag
     3841 cctgtccgcc tacgccccag cccgcctggc tgccgcagcc ctgctcctgg ccagactgac
     3901 gcacgggcag acacagccct ggaccactca gctgtgggac ctcaccggat tctcctatga
     3961 agacctcatt ccctgcgtct tgagcctcca taagaagtgc ttccatgatg acgcccccaa
     4021 ggactacagg caagtctctc tgaccgccgt gaagcagcgg tttgaggaca agcgctatgg
     4081 agaaatcagc caggaagagg tgctgagcta cagccagttg tgtgctgcat taggagtgac
     4141 acaagacagc cccgaccccc cgactttcct cagcacaggg gagatccacg ccttcctcag
     4201 ctctccctcg gggcggagaa ccaaacggaa gcgggagaac agcctccagg aagacagagg
     4261 cagcttcgtt accaccccca ctgcggagct gtccagccag gaggagacgc tgctgggcag
     4321 cttcctcgac tggagcctgg actgctgctc tggctatgaa ggcgaccagg agagtgaggg
     4381 cgagaaggag ggcgacgtga cagctcccag cggcatcctc gatgtcaccg tggtctacct
     4441 gaacccagaa cagcattgct gccaggaatc cagtgatgag gaggcttgtc cagaggacaa
     4501 gggaccccag gacccacagg cactggcgct ggacacccag atccctgcaa cccctggacc
     4561 caaacccctg gtccgcacca gccgggagcc agggaaggac gtcacgacct cagggtactc
     4621 ctccgtcagc accgcaagtc ccacaagctc cgtggacggt ggcttggggg ccctgcccca
     4681 acctacctca gtgctgtccc tggacagtga ctcgcacaca cagccctgcc accatcaggc
     4741 caggaagtca tgtttacagt gtcgtccccc aagtcccccg gagagcagtg ttccccagca
     4801 acaggtgaag cggataaacc tatgcataca cagtgaggag gaggacatga acctgggcct
     4861 tgtgaggctg taagcggccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct
     4921 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     4981 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     5041 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     5101 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     5161 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     5221 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     5281 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     5341 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     5401 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     5461 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     5521 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     5581 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     5641 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5701 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5761 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5821 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5881 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     5941 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     6001 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     6061 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     6121 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     6181 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     6241 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     6301 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     6361 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     6421 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     6481 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     6541 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     6601 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     6661 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     6721 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     6781 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     6841 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     6901 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     6961 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     7021 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     7081 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     7141 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     7201 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     7261 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     7321 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     7381 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     7441 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     7501 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     7561 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7621 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     7681 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7741 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     7801 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7861 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7921 gca
//
