LOCUS       sequence-261834-        7364 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  inducible mammalian expression vector of eGFP 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 7364)
  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 7364)
  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: pcDNA5-eGFP-FRT-T0-RNF168 delta MIU1/MIU2
FEATURES             Location/Qualifiers
     source          1..7364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(165..956,957..1025))
                     /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     788..807
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1026..1130)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1205..1224)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1396..1775
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1776..1979
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1930..1950
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    1981..1999
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2002..2020
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2030..2054
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(2151..2153,2154..2156,2157..2867)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2151..2153,2154..2156,2157..2867)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2196..2217)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2457..2476)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2804..2825
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4220..4231
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(4477..4494)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4483..4707
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4840..4859)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4991..5038
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             5046..6068
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    6198..6319
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6235..6254)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6289..6308
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6368..6384)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6368..6384)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6381..6403)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6392..6408
                     /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(6416..6422,6423..6440,6441..6446))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6461..6482
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6599..6616)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6770..7358)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6850..6869)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
       61 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
      121 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
      181 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
      241 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
      301 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
      361 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
      421 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
      481 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
      541 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
      601 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
      661 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
      721 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
      781 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
      841 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
      901 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
      961 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     1021 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     1081 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     1141 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
     1201 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     1261 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     1321 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
     1381 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
     1441 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     1501 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     1561 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     1621 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     1681 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     1741 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     1801 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     1861 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     1921 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     1981 tccctatcag tgatagagat ctccctatca gtgatagaga tcgtcgacga gctcgtttag
     2041 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
     2101 gggaccgatc cagcctccgg actctagcgt ttaaacttaa gcttggtacc atggtgagca
     2161 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     2221 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     2281 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2341 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     2401 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     2461 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2521 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2581 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     2641 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     2701 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     2761 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     2821 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc gcgcctatgg
     2881 ctctacccaa agacgccatc ccctcgctgt ccgagtgcca gtgcgggatc tgcatggaaa
     2941 tcctcgtgga gcccgtcacc ctcccgtgta accacacgct gtgtaaaccg tgcttccagt
     3001 cgaccgtcga aaaggcgagt ttatgctgtc ccttctgtcg ccgccgggta tcgtcgtgga
     3061 ctcggtacca tacccgaaga aattctctcg tcaacgtgga actgtggacg ataattcaaa
     3121 aacactatcc cagggagtgc aagcttagag cgtctggcca agaatcagag gaagtggctg
     3181 atgactatca gccagttcgt ctgctcagta aacctgggga actgagaaga gaatatgaag
     3241 aggaaataag caaggtggcg gcagagcgac gggccagcga ggaagaagaa aacaaagcca
     3301 gtgaagaata catacagagg ttgttggcag aggaggaaga agaggaaaaa agacaggcag
     3361 aaaaaaggcg aagagcggga agtatctcgg cttctccctt gaattccaga aaatctgatc
     3421 cagttacacc caagtctgaa aagaaaagta agaacaaaca aagaaacact ggagatattc
     3481 agaagtattt gacaccgaaa tctcagtttg ggtcagcctc acactctgaa gctgtacaag
     3541 aagtcaggaa agactccgta tctaaggaca ttgacagtag tgataggaaa agcccaacag
     3601 ggcaagacac agaaatagaa gatatgccga cactttctcc acagatatcc cttggagttg
     3661 gagaacaagg tgcagattct tcaatagagt cccctatgcc atggttatgt gcctgtggtg
     3721 ccgaatggta ccatgaagga aacgtcaaaa caagaccaag caatcatggg aaagagttat
     3781 gtgtcttaag tcacgagcga cctaaaacca gagttcccta ctcgaaagaa actgcagtta
     3841 tgccttgtgg cagaacagaa agtgggtgcg cccccacatc aggggtgaca cagacaaatg
     3901 gaaacaacac aggtgagaca gaaaatgaag agtcgtgcct actgatcagt aaggagattt
     3961 ccaaaagaaa aaaccaagaa tcttcctttg aagcagtcaa ggatccatgc ttttctgcaa
     4021 aaagaagaaa agtgtccccc gaatcttccc cagatcaaga ggaaacagaa ataaacttta
     4081 cccaaaaact gatagatttg gagcatctac tgtttgaggt gccaaaccgg caaaaaggat
     4141 ccccagatga gtatcactta cgcgctacat cctcccctcc agacaaagtg ctaaatggac
     4201 agaggaagaa tcccaaagat gggaacttca aaaggcaaac tcacacaaag catccaacac
     4261 cagagagagg ctcaagggac aaaaataggc aagtgtcttt aaagatgcag ttgaagcagt
     4321 cagttaatag aagaaagatg ccaaattcta ctagagatca ctgtaaggta tccaaaagtg
     4381 ctcactccct acagcctagc atttcacaga aaagtgtttt tcagatgttt cagagatgca
     4441 caaagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     4501 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     4561 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     4621 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     4681 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     4741 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4801 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4861 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4921 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4981 cacgtaccta gaagttccta ttccgaagtt cctattctct agaaagtata ggaacttcct
     5041 tggccaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg
     5101 acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg
     5161 atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag
     5221 atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca
     5281 ttggggagtt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt
     5341 tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg
     5401 atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag
     5461 gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt
     5521 atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg
     5581 agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg
     5641 gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg
     5701 cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg
     5761 cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc
     5821 cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg
     5881 acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg
     5941 gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg
     6001 gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa
     6061 aggaatagca cgtactacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     6121 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     6181 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     6241 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     6301 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     6361 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6421 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6481 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6541 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6601 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6661 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6721 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6781 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6841 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6901 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6961 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7021 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7081 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7141 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7201 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7261 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7321 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agat
//
