LOCUS       sequence-261837-        6983 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression vector of Flag tagged RNF168 where both motifs 
            interacting with ubiquitin have been deleted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6983)
  AUTHORS   Stewart GS, Panier S, Townsend K, Al-Hakim AK, Kolas NK, Miller ES, 
            Nakada S, Ylanko J, Olivarius S, Mendez M, Oldreive C, Wildenhain J,
            Tagliaferro A, Pelletier L, Taubenheim N, Durandy A, Byrd PJ, 
            Stankovic T, Taylor AM, Durocher D
  TITLE     The RIDDLE syndrome protein mediates a ubiquitin-dependent signaling
            cascade at sites of DNA damage.
  JOURNAL   Cell. 2009 Feb 6;136(3):420-34. doi: 10.1016/j.cell.2008.12.042.
  PUBMED    19203578
REFERENCE   2  (bases 1 to 6983)
  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.2008.12"; journalName: "Cell"; date: "2009-02-6- 6";
            volume: "136"; issue: "3"; pages: "420-34"
COMMENT     Sequence Label: pcDNA3-Flag-RNF168 delta MIU1/MIU2
FEATURES             Location/Qualifiers
     source          1..6983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        167..546
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        547..750
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     701..721
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     795..814
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        795..813
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             836..859
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2227..2238
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(2468..2486)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2469..2486)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2506..2523)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2512..2736
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2782..3210
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2869..2888)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3079..3100
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3219..3239)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3224..3553
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3404..3539
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3466..3485
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3620..4414
                     /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(3674..3693)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4284..4303
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4588..4709
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4625..4644)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4679..4698
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4758..4774)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4758..4774)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4771..4793)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4782..4798
                     /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(4806..4812,4813..4830,4831..4836))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4851..4872
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4989..5006)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5160..5748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5240..5259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5919..6710,6711..6779))
                     /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     6542..6561
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6780..6884)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6959..6978)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
       61 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
      121 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
      181 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      241 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      301 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      361 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      421 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      481 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      541 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      601 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      661 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      721 gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac
      781 tggcttatcg aaattaatac gactcactat agggagaccc aagcttggta ccatggacta
      841 caaagacgat gacgataaag ccgccagttt aaacggcgcg cctatggctc tacccaaaga
      901 cgccatcccc tcgctgtccg agtgccagtg cgggatctgc atggaaatcc tcgtggagcc
      961 cgtcaccctc ccgtgtaacc acacgctgtg taaaccgtgc ttccagtcga ccgtcgaaaa
     1021 ggcgagttta tgctgtccct tctgtcgccg ccgggtatcg tcgtggactc ggtaccatac
     1081 ccgaagaaat tctctcgtca acgtggaact gtggacgata attcaaaaac actatcccag
     1141 ggagtgcaag cttagagcgt ctggccaaga atcagaggaa gtggctgatg actatcagcc
     1201 agttcgtctg ctcagtaaac ctggggaact gagaagagaa tatgaagagg aaataagcaa
     1261 ggtggcggca gagcgacggg ccagcgagga agaagaaaac aaagccagtg aagaatacat
     1321 acagaggttg ttggcagagg aggaagaaga ggaaaaaaga caggcagaaa aaaggcgaag
     1381 agcgggaagt atctcggctt ctcccttgaa ttccagaaaa tctgatccag ttacacccaa
     1441 gtctgaaaag aaaagtaaga acaaacaaag aaacactgga gatattcaga agtatttgac
     1501 accgaaatct cagtttgggt cagcctcaca ctctgaagct gtacaagaag tcaggaaaga
     1561 ctccgtatct aaggacattg acagtagtga taggaaaagc ccaacagggc aagacacaga
     1621 aatagaagat atgccgacac tttctccaca gatatccctt ggagttggag aacaaggtgc
     1681 agattcttca atagagtccc ctatgccatg gttatgtgcc tgtggtgccg aatggtacca
     1741 tgaaggaaac gtcaaaacaa gaccaagcaa tcatgggaaa gagttatgtg tcttaagtca
     1801 cgagcgacct aaaaccagag ttccctactc gaaagaaact gcagttatgc cttgtggcag
     1861 aacagaaagt gggtgcgccc ccacatcagg ggtgacacag acaaatggaa acaacacagg
     1921 tgagacagaa aatgaagagt cgtgcctact gatcagtaag gagatttcca aaagaaaaaa
     1981 ccaagaatct tcctttgaag cagtcaagga tccatgcttt tctgcaaaaa gaagaaaagt
     2041 gtcccccgaa tcttccccag atcaagagga aacagaaata aactttaccc aaaaactgat
     2101 agatttggag catctactgt ttgaggtgcc aaaccggcaa aaaggatccc cagatgagta
     2161 tcacttacgc gctacatcct cccctccaga caaagtgcta aatggacaga ggaagaatcc
     2221 caaagatggg aacttcaaaa ggcaaactca cacaaagcat ccaacaccag agagaggctc
     2281 aagggacaaa aataggcaag tgtctttaaa gatgcagttg aagcagtcag ttaatagaag
     2341 aaagatgcca aattctacta gagatcactg taaggtatcc aaaagtgctc actccctaca
     2401 gcctagcatt tcacagaaaa gtgtttttca gatgtttcag agatgcacaa agtctagagg
     2461 gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt
     2521 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     2581 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     2641 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     2701 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     2761 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2821 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     2881 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2941 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3001 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3061 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3121 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3181 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     3241 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3301 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3361 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3421 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3481 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3541 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     3601 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3661 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3721 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3781 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3841 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3901 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3961 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4021 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4081 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4141 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4201 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4261 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4321 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4381 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4441 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4501 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4561 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     4621 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4681 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     4741 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4801 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4861 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4921 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4981 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5041 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5101 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5161 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5221 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5281 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5341 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5401 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5461 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5521 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5581 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5641 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5701 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5761 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5821 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5881 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5941 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6001 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6061 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6121 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6181 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6241 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6301 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6361 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6421 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6481 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6541 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6601 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6661 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6721 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6781 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6841 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6901 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     6961 caatctgctc tgatgccgca tag
//
