LOCUS       sequence-473579-        7831 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 7831)
  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 7831)
  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 HA Cyclin F
FEATURES             Location/Qualifiers
     source          1..7831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..24
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(243..347)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(422..441)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        613..992
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        993..1196
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1147..1167
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1241..1260
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1241..1259
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1282..1308
                     /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             1282..1308
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1288..1314
                     /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(3762..3780)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3763..3780)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3800..3817)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3806..4030
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4076..4504
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4163..4182)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4373..4394
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4513..4533)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4518..4847
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4698..4833
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4760..4779
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4914..5708
                     /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(4968..4987)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5578..5597
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5882..6003
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5919..5938)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5973..5992
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6052..6068)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6052..6068)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6065..6087)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6076..6092
                     /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(6100..6106,6107..6124,6125..6130))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6145..6166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6283..6300)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6454..7042)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6534..6553)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7213..7831,1..173,174..242))
                     /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"
ORIGIN
        1 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
       61 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
      121 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
      181 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
      241 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
      301 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
      361 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
      421 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
      481 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
      541 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca
      601 gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
      661 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      721 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      781 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      841 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      901 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      961 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg
     1021 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     1081 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc
     1141 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct
     1201 ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact cactataggg
     1261 agacccaagc ttaccatggc ctacccctac gacgtgcccg actacgcctc cctcggcggg
     1321 agcggcggcg tggtccactg taggtgtgcc aagtgtttct gttatcctac aaagcgaaga
     1381 ataaggagga ggccccgaaa cctgaccatc ttgagtctcc ccgaagatgt gctctttcac
     1441 atcctgaaat ggctttctgt agaggacatc ctggccgtcc gagctgtaca ctcccagctg
     1501 aaggacctgg tggacaacca cgccagtgtg tgggcatgtg ccagcttcca ggagctgtgg
     1561 ccgtctccag ggaacctgaa gctctttgaa agggctgctg aaaaggggaa tttcgaagct
     1621 gctgtgaagc tgggcatagc ctacctctac aatgaaggcc tgtctgtgtc tgatgaggcc
     1681 cgcgcagaag tgaatggcct gaaggcctct cgcttcttca gtctcgctga gcggctgaat
     1741 gtgggtgccg cacctttcat ctggctcttc atccgccctc cgtggtcggt gagcggaagc
     1801 tgctgcaagg ccgtggttca cgagagcctc agggcagagt gccagctgca gaggactcac
     1861 aaagcatcca tattgcactg cttgggcaga gtgctgagtc tgttcgagga tgaggagaag
     1921 cagcagcagg cccatgacct gtttgaggag gctgctcatc agggatgtct gaccagctcc
     1981 tacctcctct gggaaagcga caggaggaca gatgtgtcag atcctgggcg atgcctccac
     2041 agcttccgaa aactcaggga ctacgctgcc aaaggctgct gggaagcgca gctgtcttta
     2101 gccaaagcct gtgcaaatgc aaaccagctt ggactggagg tgagagcttc cagtgagatc
     2161 gtctgccagc tatttcaggc ttcccaggct gtcagtaaac aacaagtctt ctccgtgcag
     2221 aagggactca atgacacaat gaggtacatt ctgatcgact ggctggtgga agttgccacc
     2281 atgaaggact tcacaagcct gtgcctgcac ctgaccgtgg agtgtgtgga ccggtacctg
     2341 cggaggaggc tggtgccgcg gtacaggctc cagctgctgg gcatcgcctg catggtcatc
     2401 tgcacccggt ttatcagtaa agagatcctg accatccggg aggccgtatg gctcacggac
     2461 aacacttaca agtacgagga cctggtgaga atgatgggcg agatcgtctc cgccttggaa
     2521 gggaagattc gagtccccac tgtggtggat tacaaggagg tcctgctgac gctagtccct
     2581 gtggagctga gaacccagca cctgtgcagc ttcctctgcg agctctccct gctgcacacc
     2641 agcctgtccg cctacgcccc agcccgcctg gctgccgcag ccctgctcct ggccagactg
     2701 acgcacgggc agacacagcc ctggaccact cagctgtggg acctcaccgg attctcctat
     2761 gaagacctca ttccctgcgt cttgagcctc cataagaagt gcttccatga tgacgccccc
     2821 aaggactaca ggcaagtctc tctgaccgcc gtgaagcagc ggtttgagga caagcgctat
     2881 ggagaaatca gccaggaaga ggtgctgagc tacagccagt tgtgtgctgc attaggagtg
     2941 acacaagaca gccccgaccc cccgactttc ctcagcacag gggagatcca cgccttcctc
     3001 agctctccct cggggcggag aaccaaacgg aagcgggaga acagcctcca ggaagacaga
     3061 ggcagcttcg ttaccacccc cactgcggag ctgtccagcc aggaggagac gctgctgggc
     3121 agcttcctcg actggagcct ggactgctgc tctggctatg aaggcgacca ggagagtgag
     3181 ggcgagaagg agggcgacgt gacagctccc agcggcatcc tcgatgtcac cgtggtctac
     3241 ctgaacccag aacagcattg ctgccaggaa tccagtgatg aggaggcttg tccagaggac
     3301 aagggacccc aggacccaca ggcactggcg ctggacaccc agatccctgc aacccctgga
     3361 cccaaacccc tggtccgcac cagccgggag ccagggaagg acgtcacgac ctcagggtac
     3421 tcctccgtca gcaccgcaag tcccacaagc tccgtggacg gtggcttggg ggccctgccc
     3481 caacctacct cagtgctgtc cctggacagt gactcgcaca cacagccctg ccaccatcag
     3541 gccaggaagt catgtttaca gtgtcgtccc ccaagtcccc cggagagcag tgttccccag
     3601 caacaggtga agcggataaa cctatgcata cacagtgagg aggaggacat gaacctgggc
     3661 cttgtgaggc tgtaacacta gtaacggccg ccagtgtgct ggaattctgc agatatccat
     3721 cacactggcg gccgctcgag catgcatcta gagggcccta ttctatagtg tcacctaaat
     3781 gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3841 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3901 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3961 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     4021 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     4081 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4141 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4201 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4261 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4321 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4381 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4441 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4501 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4561 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4621 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4681 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4741 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4801 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4861 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4921 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4981 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     5041 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     5101 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     5161 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5221 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5281 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5341 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5401 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     5461 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5521 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5581 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5641 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5701 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5761 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5821 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5881 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5941 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6001 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     6061 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6121 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6181 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6241 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6301 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6361 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6421 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6481 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6541 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6601 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6661 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6721 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6781 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6841 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6901 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6961 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7021 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7081 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7141 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7201 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7261 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7321 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7381 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7441 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7501 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7561 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7621 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7681 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7741 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7801 gcgaccgagt tgctcttgcc cggcgtcaat a
//
