LOCUS       sequence-261816-        7838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression vector of eGFP tagged wild type RNF168 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7838)
  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 7838)
  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-eGFP-NLS-RNF168
FEATURES             Location/Qualifiers
     source          1..7838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        96..475
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        476..679
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     630..650
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     724..743
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        724..742
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(762..764,765..767,768..1478)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(762..764,765..767,768..1478)
                     /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(807..828)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1068..1087)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1415..1436
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1488..1508
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3008..3019
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(3252..3270)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3253..3270)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3290..3307)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3296..3520
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3566..3994
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3653..3672)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3863..3884
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4003..4023)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4008..4337
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4188..4323
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4250..4269
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4404..5198
                     /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(4458..4477)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5068..5087
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5372..5493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5409..5428)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5463..5482
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5542..5558)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5542..5558)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5555..5577)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5566..5582
                     /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(5590..5596,5597..5614,5615..5620))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5635..5656
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5773..5790)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5944..6532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6024..6043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6703..7494,7495..7563))
                     /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     7326..7345
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7564..7668)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7743..7762)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
       61 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
      121 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      181 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      241 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      301 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      361 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      421 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      481 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      541 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      601 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      661 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
      721 aattaatacg actcactata gggagaccca agcttggtac catggtgagc aagggcgagg
      781 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      841 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
      901 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
      961 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1021 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     1081 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1141 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     1201 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     1261 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     1321 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     1381 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1441 ccgccgggat cactctcggc atggacgagc tgtacaaggg cgcgggaccc aagaagaagc
     1501 ggaaggtggg agggcgcgcc atggctctac ccaaagacgc catcccctcg ctgtccgagt
     1561 gccagtgcgg gatctgcatg gaaatccttg tggagcccgt caccctcccg tgtaaccaca
     1621 cgctgtgtaa accgtgcttc cagtcgaccg tcgaaaaggc gagtttatgc tgtcccttct
     1681 gtcgccgccg ggtatcgtcg tggactcggt accatacccg aagaaattct ctcgtcaacg
     1741 tggaactgtg gacgataatt caaaaacact atcccaggga gtgcaagctt agagcgtctg
     1801 gccaagaatc agaggaagtg gctgatgact atcagccagt tcgtctgctc agtaaacctg
     1861 gggaactgag aagagaatat gaagaggaaa taagcaaggt ggcggcagag cgacgggcca
     1921 gcgaggaaga agaaaacaaa gccagtgaag aatacataca gaggttgttg gcagaggagg
     1981 aagaagagga aaaaagacag gcagaaaaaa ggcgaagagc gatggaagaa caactgaaaa
     2041 gtgatgagga actggcaaga aagctaagca ttgatattaa caatttctgt gagggaagta
     2101 tctcggcttc tcccttgaat tccagaaaat ctgatccagt tacacccaag tctgagaaga
     2161 aaagtaagaa caaacaaaga aacactggag atattcagaa gtatttgaca ccgaaatctc
     2221 agtttgggtc agcctcacac tctgaagctg tacaagaagt caggaaagac tccgtatcta
     2281 aggacattga cagtagtgat aggaaaagcc caacagggca agacacagaa atagaagata
     2341 tgccgacact ttctccacag atatcccttg gagttggaga acaaggtgca gattcttcaa
     2401 tagagtcccc tatgccatgg ttatgtgcct gtggtgccga atggtaccat gaaggaaacg
     2461 tcaaaacaag accaagcaat catgggaaag agttatgtgt cttaagtcac gagcgaccta
     2521 aaaccagagt tccctactcg aaagaaactg cagttatgcc ttgtggcaga acagaaagtg
     2581 ggtgcgcccc cacatcaggg gtgacacaga caaatggaaa caacacaggt gagacagaaa
     2641 atgaggagtc gtgtttattg atcagtaagg agatttccaa aagaaaaaac caagaatctt
     2701 cctttgaagc agtcaaggat ccatgctttt ctgcaaaaag aagaaaagtg tcccccgaat
     2761 cttccccaga tcaagaggaa acagaaataa actttaccca aaaactgata gatttggagc
     2821 atctactgtt tgagagacat aaacaagaag aacaggacag gttattggca ttacaacttc
     2881 agaaggaggt ggataaagag caaatggtgc caaaccggca aaaaggatcc ccagatgagt
     2941 atcacttacg cgctacatcc tcccctccag acaaagtgct aaatggacag aggaagaatc
     3001 ccaaagatgg gaacttcaaa aggcaaactc acacaaagca tccaacacca gagagaggct
     3061 caagggacaa aaataggcaa gtgtctttaa agatgcagtt gaagcagtca gttaatagaa
     3121 gaaagatgcc aaattctact agagatcact gtaaggtatc aaaaagtgct cactccctac
     3181 agcctagcat ttcacagaaa agtgtttttc agatgtttca gagatgcaca aagtaatcta
     3241 gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg
     3301 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3361 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3421 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3481 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3541 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3601 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3661 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3721 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3781 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3841 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3901 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3961 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4021 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4081 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4141 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4201 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4261 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4321 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     4381 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4441 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4501 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4561 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4621 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4681 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4741 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4801 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4861 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4921 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4981 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5041 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5101 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5161 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5221 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5281 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5341 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5401 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5461 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5521 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5581 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5641 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5701 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5761 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5821 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5881 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5941 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6001 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6061 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6121 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6181 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6241 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6301 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6361 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6421 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6481 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6541 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6601 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6661 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6721 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6781 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6841 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6901 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6961 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7021 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7081 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7141 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7201 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7261 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7321 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7381 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7441 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7501 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7561 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7621 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7681 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
     7741 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     7801 gaggtcgctg agtagtgcgc gagcaaaatt taagctac
//
