LOCUS       sequence-473582-        6575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transient overexpression of RRM2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6575)
  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 6575)
  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 FLAG RRM2
FEATURES             Location/Qualifiers
     source          1..6575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..30)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     257..277
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(286..714)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(396..417)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     608..627
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    760..984
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     973..990
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2210..2233)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2274..2293)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2275..2293)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2338..2541)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2367..2387)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2542..2921
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3093..3112
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3187..3291
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3292..3360,3361..4152)
                     /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(3510..3529)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4323..4911
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4812..4831
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5065..5082
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5199..5220
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5235..5240,5241..5258,5259..5265)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5273..5289
                     /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     5278..5300
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5297..5313
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5297..5313
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5362..5483
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5373..5392)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5427..5446
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5657..6451)
                     /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(5768..5787)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6378..6397
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6518..6575,1..272))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6532..6575,1..92)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gcagaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa
       61 tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt
      121 gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt
      181 ccacacctgg ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag
      241 cctggggact ttccacaccc taactgacac acattccaca gaattaattc gcgttaaatt
      301 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
      361 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
      421 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
      481 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
      541 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
      601 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
      661 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtggggat
      721 accccctaga gccccagctg gttctttccg cctcagaagc catagagccc accgcatccc
      781 cagcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc accccacccc
      841 ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt tattaggaaa
      901 ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg caaacaacag
      961 atggctggca actagaaggc acagtcgagg ctgatcagcg ggtttaaacg ggccctctag
     1021 actcgagtta gaagtcagca tccaaggtaa aagaattctc tgttggactt gacatcactc
     1081 ccatcctctg atactcgcct actctcttct caaagaagtt agtctttcct tccagtgaaa
     1141 tattctccat aaagtcaaat gggttctcta ctctgaaaac cttgctaaaa cccagttcca
     1201 gcataagtct gtctgccaca aactcaatgt attgcttcat tagagtgcaa ttcatcccaa
     1261 tgagcttcac aggcaaggcc tcagtgagga actcctgttc tatccgaaca gcattgataa
     1321 ttatttctct tactctctcc tccgatggtt tgtgtaccag gtgtttgaac atcaggcaag
     1381 caaaatcaca gtgtaaaccc tcatctctgc taataagttc attagaaaat gtgaggccag
     1441 gcatcagtcc tcgtttcttg agccagaata tcgacgcaaa agaaccggaa aagaaaatgc
     1501 cttccactgc agcaaaggct acaacacgtt caccataggt agcctctttg tccccaatcc
     1561 agcgcaaggc ccagtctgcc ttcttcttga cacaaggcat cgtttcaatg gcattgaaga
     1621 gaaattccct ttctttggga tcttttatgt aagtgtcaat aagaagacta tacatttcag
     1681 aatgtatgtt ttccatggca atttggaagc catagaaaca gcgggcttct gtaatctgaa
     1741 cttcttggct aaatcgctcc accaagtttt catttactat gccatcgctt gctgcaaaga
     1801 aagccagaac atgggatata aaatatctct cctcgggttt cagggattcc cagtgctgaa
     1861 tgtccttgga gaggtccacc tcctcggcgg tccaaaagga agcctctgcc ttcttataca
     1921 tctgccagat atcatggtac tcgatgggga agatgacaaa gcggcggggg ttttctctca
     1981 gcagcggctc atcctccacg ccgggggcag ctgctttagt tttcggctcc gtgggctcct
     2041 ggaagatcct cctcgcggtc ttgctggcca ggacgcgggt cccgctcagg gccggcggcg
     2101 tgttctcctt gtcgaccaag ctgagcccct tcagcggcga gagctgcagc tgctgcgggt
     2161 ccgtgatggg cgcgagcggg acacggaggg agagcatgga tcctggtacc ttgtcatcgt
     2221 cgtccttgta gtccatggta agcttaagtt taaacgctag ccagcttggg tctccctata
     2281 gtgagtcgta ttaatttcga taagccagta agcagtgggt tctctagtta gccagagagc
     2341 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga
     2401 gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     2461 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     2521 gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca
     2581 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     2641 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     2701 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     2761 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     2821 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     2881 cgtaattgat tactattaat aactagtcaa taatcaatgt caacgcgtat atctggcccg
     2941 tacatcgcga agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc aattgtcggt
     3001 caagccttgc cttgttgtag cttaaatttt gctcgcgcac tactcagcga cctccaacac
     3061 acaagcaggg agcagatact ggcttaacta tgcggcatca gagcagattg tactgagagt
     3121 gcaccatagg ggatcgggag atctcccgat ccgtcgacgt caggtggcac ttttcgggga
     3181 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3241 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3301 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3361 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3421 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3481 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3541 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3601 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3661 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3721 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3781 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3841 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3901 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3961 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4021 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4081 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4141 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4201 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4261 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4321 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4381 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4441 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4501 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4561 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4621 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4681 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4741 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4801 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4861 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4921 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     4981 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5041 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     5101 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     5161 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     5221 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     5281 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag
     5341 gtcgacggta tacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     5401 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     5461 ttataagctg caataaacaa gttggggtgg gcgaagaact ccagcatgag atccccgcgc
     5521 tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag
     5581 gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg
     5641 aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     5701 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     5761 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     5821 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     5881 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg agcctggcga
     5941 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     6001 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     6061 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     6121 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     6181 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     6241 ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg
     6301 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     6361 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     6421 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     6481 cagatccgaa aatggatata caagctcccg ggagcttttt gcaaaagcct aggcctccaa
     6541 aaaagcctcc tcactacttc tggaatagct cagag
//
