LOCUS       sequence-436411-        7269 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses RNF169 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7269)
  AUTHORS   Panier S, Ichijima Y, Fradet-Turcotte A, Leung CC, Kaustov L, 
            Arrowsmith CH, Durocher D
  TITLE     Tandem protein interaction modules organize the ubiquitin-dependent 
            response to DNA double-strand breaks.
  JOURNAL   Mol Cell. 2012 Aug 10;47(3):383-95. doi: 
            10.1016/j.molcel.2012.05.045. Epub 2012 Jun 27.
  PUBMED    22742833
REFERENCE   2  (bases 1 to 7269)
  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.molcel.2012.05.045"; journalName: "Mol Cell"; date: 
            "2012-08-10- 10"; volume: "47"; issue: "3"; pages: "383-95"
COMMENT     Sequence Label: pcDNA5-FRT/TO-Flag-RNF169
FEATURES             Location/Qualifiers
     source          1..7269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1040..1161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1051..1070)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1105..1124
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1291..2313)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(2321..2368)
                     /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)."
     primer_bind     2500..2519
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2652..2876
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2865..2882
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(5087..5110)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5210..5234)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    5244..5262
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5265..5283
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(5285..5488)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5314..5334)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5489..5868
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6040..6059
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6134..6238
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6239..6307,6308..7099)
                     /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     complement(6457..6476)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
     1021 cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1081 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1141 ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg
     1201 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
     1261 ggcggtggaa tcgaaatctc gtagtacgtg ctattccttt gccctcggac gagtgctggg
     1321 gcgtcggttt ccactatcgg cgagtacttc tacacagcca tcggtccaga cggccgcgct
     1381 tctgcgggcg atttgtgtac gcccgacagt cccggctccg gatcggacga ttgcgtcgca
     1441 tcgaccctgc gcccaagctg catcatcgaa attgccgtca accaagctct gatagagttg
     1501 gtcaagacca atgcggagca tatacgcccg gagccgcggc gatcctgcaa gctccggatg
     1561 cctccgctcg aagtagcgcg tctgctgctc catacaagcc aaccacggcc tccagaagaa
     1621 gatgttggcg acctcgtatt gggaatcccc gaacatcgcc tcgctccagt caatgaccgc
     1681 tgttatgcgg ccattgtccg tcaggacatt gttggagccg aaatccgcgt gcacgaggtg
     1741 ccggacttcg gggcagtcct cggcccaaag catcagctca tcgagagcct gcgcgacgga
     1801 cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac acatggggat cagcaatcgc
     1861 gcatatgaaa tcacgccatg tagtgtattg accgattcct tgcggtccga atgggccgaa
     1921 cccgctcgtc tggctaagat cggccgcagc gatcgcatcc atggcctccg cgaccggctg
     1981 cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac gtgacaccct gtgcacggcg
     2041 ggagatgcaa taggtcaggc tctcgctgaa ctccccaatg tcaagcactt ccggaatcgg
     2101 gagcgcggcc gatgcaaagt gccgataaac ataacgatct ttgtagaaac catcggcgca
     2161 gctatttacc cgcaggacat atccacgccc tcctacatcg aagctgaaag cacgagattc
     2221 ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg aacttttcga tcagaaactt
     2281 ctcgacagac gtcgcggtga gttcaggctt tttggccaag gaagttccta tactttctag
     2341 agaataggaa cttcggaata ggaacttcta ggtacgtgaa ccatcaccct aatcaagttt
     2401 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     2461 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     2521 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     2581 gcttaatgcg ccgctacagg gcgcgtgggg atacccccta gagccccagc tggttctttc
     2641 cgcctcagaa gccatagagc ccaccgcatc cccagcatgc ctgctattgt cttcccaatc
     2701 ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca
     2761 atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt
     2821 caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga
     2881 ggctgatcag cgggtttaaa cgggccctct agactcgagc ggccgctcac ttggccccgg
     2941 ccatgttgct ggaccgtagg agatactgat ccacacttcc ttttcgccgg ctcacagtcc
     3001 gcctctcatt gtcgaacatg cgctgcaact gcagagccaa ctgtcggtct tcttcctctt
     3061 gctgaagctt ctgctccatc tctcgcagga ctgggtctgt tggggccaga cccacctccc
     3121 cactggtttg tcgcagtttt ttaagggagc cattttgttc taagtgcttg gtcttgcagt
     3181 gtcttttccg gcctcgacgc aaagaaggaa gtggctcttc acttaggctc tcaactagaa
     3241 caccattagt cagatcaaaa tgatttaatg tcttcaattg ttgctttgtt ttgaggactc
     3301 cacccaaaac actgttttgg ggtagcactg aattaactgt ggtgattttc atggctctgc
     3361 ttatacaggt tttgtctaac ttggcatctg gagttgaccc taacccctca aactgctccc
     3421 tctccaaaga agtcccactg cctccccctt tgagttctga ggaacagcag gtttccagtg
     3481 ggatctcagt gctactttta ttatcactgt cctgttctgc ttttgtttgg ctaacagagg
     3541 ggaaatgatc aggatcagca gaggtgggtc cagtatactc agagaggacc tgcccaccag
     3601 ataatcttgt atttacagcc acaagtggct tctccttgct agaatggata ccttcagagc
     3661 ctagtaactc ttcccccatt tcaggagcca gagaggtaag agtggctttt gaaagggtct
     3721 ttttgatctg ccgctcctga aagatctgtt cccacttttt gaggatccgt ggactggcct
     3781 cataagaagt ctgcttttgc aggcttctgt ttaggttgcg tggagttgat ttgatgatga
     3841 gaggacttag cacacggcca tcagggagtc tcttgggagg agtacatggt gagcagacaa
     3901 tgggcttgaa atggtttagt tcttcggaga tgctgtcatt gctctcaggg ctgacagaac
     3961 gctctggctt atgcagggaa gccaaggatg aaagggagct gaagggtaga cgcttttcga
     4021 tggttaagtc aggggccgag acgcagcggt tgttttgagt tgacaagagg acaccaatga
     4081 tggggttgga ggctggagtc atagttgtga ccttggcttt gttgcctgga actgctctga
     4141 ctctgctctt cgctctttcc tgggctgtgt cactacagct ctgactcctt tccaccttgg
     4201 agtttagctt ccctgccagg aaagctaagg agtaggagtt gttcttggaa acaaatgccg
     4261 agcgatgtgt ctgtgtcatc tggcctcgag aaggttcttc attctctgaa tctgagagac
     4321 gtgcaggaca acgttccaga tttgttttca gtactaatgg ttcatctctt tttttctgtt
     4381 catccatttt ccttttccct gtttctgtat cctctggtaa cagcttgtgg atttgatctt
     4441 cagagggttt ttcctcttgt aacttttctt ctctcagctt tctcaagctt tcatattcct
     4501 cacgaagttc cccaggcttg cttaatttga ttggtgctct gaatataaag tctggctccg
     4561 caggcgcggc acgcggctcc tgctctggcc tgggccccgc ggcagccgcg cccccgtccc
     4621 ggcgcgggcg gcagcgctcg ggttggctgc ggcgggcgca ctcgcccagc acctctgagt
     4681 cggcctggcc gtcgtcgcgg gcccgacggc gggcccagcc tgggccgcgg gcgcggcagc
     4741 gagggcaacc cgggcccgcc gcgtcggcgg cgcgttgggc gcagcctcgg caaagcgagt
     4801 ggccgcacgg cagggccgct gcttctccgg ggggctccag gcacccggcg cagcccgatt
     4861 cctccggccg cggcggcagc ggcggctgca gcaacggcgg cgacaacacc aacagcgaag
     4921 gtccagaagc cgggcctggg gccccagtct tagctgccgc cgtctcgtca cagcggcccc
     4981 gccggccccg ccgactcaga gcggctgctg ccgccgcgga agaggcccga gtactcggac
     5041 ctgcagccgc cattttaatt aatggcgcgc cgtttaaact ggcggcttta tcgtcatcgt
     5101 ctttgtagtc catggtacca agcttaagtt taaacgctag agtccggagg ctggatcggt
     5161 cccggtgtct tctatggagg tcaaaacagc gtggatggcg tctccaggcg atctgacggt
     5221 tcactaaacg agctcgtcga cgatctctat cactgatagg gagatctcta tcactgatag
     5281 ggagagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
     5341 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
     5401 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
     5461 cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt
     5521 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     5581 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     5641 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     5701 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     5761 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     5821 atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcaa cgcgtatatc
     5881 tggcccgtac atcgcgaagc agcgcaaaac gcctaaccct aagcagattc ttcatgcaat
     5941 tgtcggtcaa gccttgcctt gttgtagctt aaattttgct cgcgcactac tcagcgacct
     6001 ccaacacaca agcagggagc agatactggc ttaactatgc ggcatcagag cagattgtac
     6061 tgagagtgca ccatagggga tcgggagatc tcccgatccg tcgacgtcag gtggcacttt
     6121 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     6181 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     6241 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6301 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6361 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6421 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6481 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6541 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6601 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6661 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6721 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6781 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6841 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6901 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6961 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     7021 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     7081 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     7141 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     7201 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7261 aggatcttc
//
