LOCUS       sequence-262016-        7055 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression vector of Flag tagged RNF168 where the second 
            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 MIU2
FEATURES             Location/Qualifiers
     source          1..7055
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        93..472
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        473..676
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     627..647
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     721..740
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        721..739
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             762..785
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2225..2236
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(2466..2484)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2467..2484)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2504..2521)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2510..2734
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2780..3208
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2867..2886)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3077..3098
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3217..3237)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3222..3551
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3402..3537
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3464..3483
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3618..4412
                     /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(3672..3691)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4282..4301
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4586..4707
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4623..4642)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4677..4696
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4756..4772)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4756..4772)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4769..4791)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4780..4796
                     /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(4804..4810,4811..4828,4829..4834))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4849..4870
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4987..5004)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5158..5746)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5238..5257)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5917..6708,6709..6777))
                     /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     6540..6559
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6778..6882)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6957..6976)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
       61 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
      121 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      181 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      241 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      301 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      361 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      421 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      481 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      541 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      601 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      661 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
      721 taatacgact cactataggg agacccaagc ttggtaccat ggactacaaa gacgatgacg
      781 ataaagccgc cagtttaaac ggcgcgccta tggctctacc caaagacgcc atcccctcgc
      841 tgtccgagtg ccagtgcggg atctgcatgg aaatcctcgt ggagcccgtc accctcccgt
      901 gtaaccacac gctgtgtaaa ccgtgcttcc agtcgaccgt cgaaaaggcg agtttatgct
      961 gtcccttctg tcgccgccgg gtatcgtcgt ggactcggta ccatacccga agaaattctc
     1021 tcgtcaacgt ggaactgtgg acgataattc aaaaacacta tcccagggag tgcaagctta
     1081 gagcgtctgg ccaagaatca gaggaagtgg ctgatgacta tcagccagtt cgtctgctca
     1141 gtaaacctgg ggaactgaga agagaatatg aagaggaaat aagcaaggtg gcggcagagc
     1201 gacgggccag cgaggaagaa gaaaacaaag ccagtgaaga atacatacag aggttgttgg
     1261 cagaggagga agaagaggaa aaaagacagg cagaaaaaag gcgaagagcg atggaagaac
     1321 aactgaaaag tgatgaggaa ctggcaagaa agctaagcat tgatattaac aatttctgtg
     1381 agggaagtat ctcggcttct cccttgaatt ccagaaaatc tgatccagtt acacccaagt
     1441 ctgaaaagaa aagtaagaac aaacaaagaa acactggaga tattcagaag tatttgacac
     1501 cgaaatctca gtttgggtca gcctcacact ctgaagctgt acaagaagtc aggaaagact
     1561 ccgtatctaa ggacattgac agtagtgata ggaaaagccc aacagggcaa gacacagaaa
     1621 tagaagatat gccgacactt tctccacaga tatcccttgg agttggagaa caaggtgcag
     1681 attcttcaat agagtcccct atgccatggt tatgtgcctg tggtgccgaa tggtaccatg
     1741 aaggaaacgt caaaacaaga ccaagcaatc atgggaaaga gttatgtgtc ttaagtcacg
     1801 agcgacctaa aaccagagtt ccctactcga aagaaactgc agttatgcct tgtggcagaa
     1861 cagaaagtgg gtgcgccccc acatcagggg tgacacagac aaatggaaac aacacaggtg
     1921 agacagaaaa tgaagagtcg tgcctactga tcagtaagga gatttccaaa agaaaaaacc
     1981 aagaatcttc ctttgaagca gtcaaggatc catgcttttc tgcaaaaaga agaaaagtgt
     2041 cccccgaatc ttccccagat caagaggaaa cagaaataaa ctttacccaa aaactgatag
     2101 atttggagca tctactgttt gaggtgccaa accggcaaaa aggatcccca gatgagtatc
     2161 acttacgcgc tacatcctcc cctccagaca aagtgctaaa tggacagagg aagaatccca
     2221 aagatgggaa cttcaaaagg caaactcaca caaagcatcc aacaccagag agaggctcaa
     2281 gggacaaaaa taggcaagtg tctttaaaga tgcagttgaa gcagtcagtt aatagaagaa
     2341 agatgccaaa ttctactaga gatcactgta aggtatccaa aagtgctcac tccctacagc
     2401 ctagcatttc acagaaaagt gtttttcaga tgtttcagag atgcacaaag tctagagggc
     2461 cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct
     2521 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2581 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     2641 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     2701 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     2761 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2821 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2881 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2941 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3001 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3061 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3121 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3181 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3241 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3301 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3361 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3421 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3481 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3541 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     3601 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3661 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3721 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3781 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3841 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3901 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3961 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4021 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4081 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4141 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4201 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4261 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4321 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4381 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4441 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4501 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4561 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4621 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4681 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4741 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4801 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4861 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4921 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4981 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5041 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5101 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5161 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5221 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5281 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5341 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5401 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5461 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5521 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5581 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5641 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5701 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5761 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5821 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5881 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5941 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6001 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6061 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6121 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6181 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6241 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6301 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6361 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6421 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6481 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6541 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6601 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6661 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6721 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6781 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6841 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6901 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     6961 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     7021 gctgagtagt gcgcgagcaa aatttaagct acaac
//
