LOCUS       sequence-261823-        7055 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression vector of Flag tagged RNF168 where the first 
            motif interacting with ubiquitin (MIU) has been deleted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7055)
  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 7055)
  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
FEATURES             Location/Qualifiers
     source          1..7055
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(12..31)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        203..582
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        583..786
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     737..757
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     831..850
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        831..849
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             872..895
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2335..2346
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(2576..2594)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2577..2594)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2614..2631)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2620..2844
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2890..3318
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2977..2996)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3187..3208
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3327..3347)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3332..3661
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3512..3647
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3574..3593
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3728..4522
                     /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(3782..3801)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4392..4411
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4696..4817
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4733..4752)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4787..4806
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4866..4882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4866..4882)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4879..4901)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4890..4906
                     /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(4914..4920,4921..4938,4939..4944))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4959..4980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5097..5114)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5268..5856)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5348..5367)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6027..6818,6819..6887))
                     /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     6650..6669
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6888..6992)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
       61 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
      121 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      181 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      241 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      301 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      361 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      421 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      481 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      541 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      601 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      661 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      721 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      781 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
      841 cactataggg agacccaagc ttggtaccat ggactacaaa gacgatgacg ataaagccgc
      901 cagtttaaac ggcgcgccta tggctctacc caaagacgcc atcccctcgc tgtccgagtg
      961 ccagtgcggg atctgcatgg aaatcctcgt ggagcccgtc accctcccgt gtaaccacac
     1021 gctgtgtaaa ccgtgcttcc agtcgaccgt cgaaaaggcg agtttatgct gtcccttctg
     1081 tcgccgccgg gtatcgtcgt ggactcggta ccatacccga agaaattctc tcgtcaacgt
     1141 ggaactgtgg acgataattc aaaaacacta tcccagggag tgcaagctta gagcgtctgg
     1201 ccaagaatca gaggaagtgg ctgatgacta tcagccagtt cgtctgctca gtaaacctgg
     1261 ggaactgaga agagaatatg aagaggaaat aagcaaggtg gcggcagagc gacgggccag
     1321 cgaggaagaa gaaaacaaag ccagtgaaga atacatacag aggttgttgg cagaggagga
     1381 agaagaggaa aaaagacagg cagaaaaaag gcgaagagcg ggaagtatct cggcttctcc
     1441 cttgaattcc agaaaatctg atccagttac acccaagtct gaaaagaaaa gtaagaacaa
     1501 acaaagaaac actggagata ttcagaagta tttgacaccg aaatctcagt ttgggtcagc
     1561 ctcacactct gaagctgtac aagaagtcag gaaagactcc gtatctaagg acattgacag
     1621 tagtgatagg aaaagcccaa cagggcaaga cacagaaata gaagatatgc cgacactttc
     1681 tccacagata tcccttggag ttggagaaca aggtgcagat tcttcaatag agtcccctat
     1741 gccatggtta tgtgcctgtg gtgccgaatg gtaccatgaa ggaaacgtca aaacaagacc
     1801 aagcaatcat gggaaagagt tatgtgtctt aagtcacgag cgacctaaaa ccagagttcc
     1861 ctactcgaaa gaaactgcag ttatgccttg tggcagaaca gaaagtgggt gcgcccccac
     1921 atcaggggtg acacagacaa atggaaacaa cacaggtgag acagaaaatg aagagtcgtg
     1981 cctactgatc agtaaggaga tttccaaaag aaaaaaccaa gaatcttcct ttgaagcagt
     2041 caaggatcca tgcttttctg caaaaagaag aaaagtgtcc cccgaatctt ccccagatca
     2101 agaggaaaca gaaataaact ttacccaaaa actgatagat ttggagcatc tactgtttga
     2161 gagacataaa caagaagaac aggacaggtt attggcatta caacttcaga aggaggtgga
     2221 taaagagcaa atggtgccaa accggcaaaa aggatcccca gatgagtatc acttacgcgc
     2281 tacatcctcc cctccagaca aagtgctaaa tggacagagg aagaatccca aagatgggaa
     2341 cttcaaaagg caaactcaca caaagcatcc aacaccagag agaggctcaa gggacaaaaa
     2401 taggcaagtg tctttaaaga tgcagttgaa gcagtcagtt aatagaagaa agatgccaaa
     2461 ttctactaga gatcactgta aggtatccaa aagtgctcac tccctacagc ctagcatttc
     2521 acagaaaagt gtttttcaga tgtttcagag atgcacaaag tctagagggc cctattctat
     2581 agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     2641 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     2701 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     2761 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     2821 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     2881 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     2941 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3001 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3061 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3121 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3181 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3241 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3301 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3361 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3421 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3481 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3541 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3601 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3661 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     3721 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3781 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3841 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3901 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3961 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4021 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4081 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4141 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4201 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     4261 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4321 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4381 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4441 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4501 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4561 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4621 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4681 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4741 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4801 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     4861 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4921 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4981 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5041 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5101 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5161 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5221 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5281 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5341 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5401 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5461 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5521 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5581 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5641 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5701 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5761 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5821 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5881 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5941 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6001 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6061 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6121 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6181 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6241 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6301 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6361 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6421 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6481 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6541 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6601 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6661 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6721 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6781 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6841 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6901 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6961 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7021 gtcgacggat cgggagatct cccgatcccc tatgg
//
