LOCUS       sequence-261841-        6953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression vector of Flag tagged RNF168 where the 
            N-terminus of RNF168 has been deleted (including RING domain).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6953)
  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 6953)
  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 RING
FEATURES             Location/Qualifiers
     source          1..6953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..498
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        499..702
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     653..673
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     747..766
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        747..765
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             788..811
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2149..2160
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(2390..2408)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2391..2408)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2428..2445)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2434..2658
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2704..3132
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2791..2810)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3001..3022
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3141..3161)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3146..3475
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3326..3461
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3388..3407
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3542..4336
                     /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(3596..3615)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4206..4225
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4510..4631
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4547..4566)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4601..4620
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4680..4696)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4680..4696)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4693..4715)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4704..4720
                     /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(4728..4734,4735..4752,4753..4758))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4773..4794
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4911..4928)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5082..5670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5162..5181)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5841..6632,6633..6701))
                     /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     6464..6483
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6702..6806)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6881..6900)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
       61 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
      121 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      181 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      241 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      301 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      361 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      421 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      481 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      541 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      601 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      661 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa
      721 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagcttgg
      781 taccatggac tacaaagacg atgacgataa agccgccagt ttaaacggcg cgccatcgtc
      841 gtggactcgg taccataccc gaagaaattc tctcgtcaac gtggaactgt ggacgataat
      901 tcaaaaacac tatcccaggg agtgcaagct tagagcgtct ggccaagaat cagaggaagt
      961 ggctgatgac tatcagccag ttcgtctgct cagtaaacct ggggaactga gaagagaata
     1021 tgaagaggaa ataagcaagg tggcggcaga gcgacgggcc agcgaggaag aagaaaacaa
     1081 agccagtgaa gaatacatac agaggttgtt ggcagaggag gaagaagagg aaaaaagaca
     1141 ggcagaaaaa aggcgaagag cgatggaaga acaactgaaa agtgatgagg aactggcaag
     1201 aaagctaagc attgatatta acaatttctg tgagggaagt atctcggctt ctcccttgaa
     1261 ttccagaaaa tctgatccag ttacacccaa gtctgagaag aaaagtaaga acaaacaaag
     1321 aaacactgga gatattcaga agtatttgac accgaaatct cagtttgggt cagcctcaca
     1381 ctctgaagct gtacaagaag tcaggaaaga ctccgtatct aaggacattg acagtagtga
     1441 taggaaaagc ccaacagggc aagacacaga aatagaagat atgccgacac tttctccaca
     1501 gatatccctt ggagttggag aacaaggtgc agattcttca atagagtccc ctatgccatg
     1561 gttatgtgcc tgtggtgccg aatggtacca tgaaggaaac gtcaaaacaa gaccaagcaa
     1621 tcatgggaaa gagttatgtg tcttaagtca cgagcgacct aaaaccagag ttccctactc
     1681 gaaagaaact gcagttatgc cttgtggcag aacagaaagt gggtgcgccc ccacatcagg
     1741 ggtgacacag acaaatggaa acaacacagg tgagacagaa aatgaagagt cgtgcctact
     1801 gatcagtaag gagatttcca aaagaaaaaa ccaagaatct tcctttgaag cagtcaagga
     1861 tccatgcttt tctgcaaaaa gaagaaaagt gtcccccgaa tcttccccag atcaagagga
     1921 aacagaaata aactttaccc aaaaactgat agatttggag catctactgt ttgagagaca
     1981 taaacaagaa gaacaggaca ggttattggc attacaactt cagaaggagg tggataaaga
     2041 gcaaatggtg ccaaaccggc aaaaaggatc cccagatgag tatcacttac gcgctacatc
     2101 ctcccctcca gacaaagtgc taaatggaca gaggaagaat cccaaagatg ggaacttcaa
     2161 aaggcaaact cacacaaagc atccaacacc agagagaggc tcaagggaca aaaataggca
     2221 agtgtcttta aagatgcagt tgaagcagtc agttaataga agaaagatgc caaattctac
     2281 tagagatcac tgtaaggtat caaaaagtgc tcactcccta cagcctagca tttcacagaa
     2341 aagtgttttt cagatgtttc agagatgcac aaagtctaga gggccctatt ctatagtgtc
     2401 acctaaatgc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     2461 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     2521 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     2581 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     2641 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     2701 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2761 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2821 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2881 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     2941 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3001 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3061 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3121 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3181 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3241 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3301 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3361 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     3421 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3481 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     3541 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3601 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3661 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3721 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3781 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3841 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3901 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3961 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4021 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4081 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4141 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4201 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4261 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4321 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4381 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4441 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4501 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4561 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4621 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     4681 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4741 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4801 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4861 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4921 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4981 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5041 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5101 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5161 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5221 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5281 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5341 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5401 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5461 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5521 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5581 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5641 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5701 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5761 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5821 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5881 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5941 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6001 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6061 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6121 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6181 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6241 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6301 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6361 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6421 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6481 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6541 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6601 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6661 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6721 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6781 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     6841 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     6901 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tag
//
