LOCUS       sequence-190609-        8154 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression construct for S protein-Flag-Streptavidin 
            binding peptide (SFB)-tagged RNF169.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8154)
  AUTHORS   Chen J, Feng W, Jiang J, Deng Y, Huen MS
  TITLE     Ring finger protein RNF169 antagonizes the ubiquitin-dependent 
            signaling cascade at sites of DNA damage.
  JOURNAL   J Biol Chem. 2012 Aug 10;287(33):27715-22. doi: 
            10.1074/jbc.M112.373530. Epub 2012 Jun 25.
  PUBMED    22733822
REFERENCE   2  (bases 1 to 8154)
  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: 
            "10.1074/jbc.M112.373530"; journalName: "J Biol Chem"; date: 
            "2012-08-10- 10"; volume: "287"; issue: "33"; pages: "27715-22"
COMMENT     Sequence Label: pMH-SFB-RNF169
FEATURES             Location/Qualifiers
     source          1..8154
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     enhancer        11..314
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        315..518
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     469..489
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             578..622
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             656..679
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             683..706
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             716..829
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     protein_bind    902..926
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2773..2797)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2980..2997)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     complement(3059..3079)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3064..3393
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3244..3379
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3403..4002
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3403..3422)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3899..3919
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4223..4692
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4822..4839)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4993..5581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5073..5092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5741..6532,6533..6601))
                     /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     6364..6383
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6602..6706)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             6737..7206
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    7271..7470
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             7671..8087
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     8017..8039
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     8055..8071
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
       61 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      121 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      181 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      241 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      301 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      361 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      421 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      481 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
      541 gctagcgcta ccggactcag atctcgaggc caccatgaaa gaaaccgctg ctgctaaatt
      601 cgaacgccag cacatggaca gcggagccgg cgcaggagcc ggcgctgacg cgcctgacta
      661 caaagacgat gacgacaagg gagattacaa ggatgacgat gacaaaggcg cgtcgatgga
      721 cgagaagacc accggctggc ggggcggcca cgtggtggag ggcctggccg gcgagctgga
      781 gcagctgcgg gccaggctgg agcaccaccc tcagggccag cgggagccct ccggcggctg
      841 caagctgggc ggcgcgcctg aattctgcag tcgacggtac cgcgggccca tcgctcgaga
      901 gacaagtttg tacaaaaaag caggcttcat ggcggctgca ggtccgagta ctcgggcctc
      961 ttccgcggcg gcagcagccg ctctgagtcg gcggggccgg cggggccgct gtgacgagac
     1021 ggcggcagct aagactgggg ccccaggccc ggcttctgga ccttcgctgt tggtgttggg
     1081 acgggggcgc ggctgccgcg gggcccaggc cagagcagga gccgcgtgcc gcgcctgcgg
     1141 agccagactt tatattcaga gcaccaatca aattaagcaa gcctggggaa cttcgtgagg
     1201 aatatgaaag cttgagaaag ctgagagaag aaaagttaca agaggaaaaa ccctctgaag
     1261 atcaaatcca caagctgtta ccagaggata cagaaacagg gaaaaggaaa atggatgaac
     1321 agaaaaaaag agatgaacca ttagtactga aaacaaatct ggaacgttgt cctgcacgtc
     1381 tctcagattc agagaatgaa gaaccttctc gaggccagat gacacagaca catcgctcgg
     1441 catttgtttc caagaacaac tcctactcct tagctttcct ggcagggaag ctaaactcca
     1501 aggtggaaag gagtcagagc tgtagtgaca cagcccagga aagagcgaag agcagagtca
     1561 gagcagttcc aggcaacaaa gccaaggtca caactatgac tccagcctcc aaccccatca
     1621 ttggtgtcct cttgtcaact caaaacaacc gctgcgtctc ggcccctgac ttaaccatcg
     1681 aaaagcgtct acccttcagc tccctttcat ccttggcttc cctgcataag ccagagcgtt
     1741 ctgtcagccc tgagagcaat gacagcatct ccgaagaact aaaccatttc aagcccattg
     1801 tctgctcacc atgtactcct cccaagagac tccctgatgg ccgtgtgcta agtcctctca
     1861 tcatcaaatc aactccacgc aacctaaaca gaagcctgca gaagcagact tcttatgagg
     1921 ccagtccacg gatcctcaaa aagtgggaac agatctttca ggagcggcag atcaaaaaga
     1981 ccctttcaaa agccactctt acctctctgg ctcctgaaat gggggaagag ttactaggct
     2041 ctgaaggtat ccattctagc aaggagaagc cacttgtggc tgtaaataca agattatctg
     2101 gtgggcaggt cctctctgag tatactggac ccacctctgc tgatcttgat catttcccct
     2161 ctgttagcca aacaaaagca gaacaggaca gtgataataa aagtagcact gagatcccac
     2221 tggaaacctg ctgttcctca gaactcaaag ggggaggcag tgggacttct ttggagaggg
     2281 agcagtttga ggggttaggg tcaactccag atgccaagtt agacaaaacc tgtataagca
     2341 gagccatgaa aatcaccaca gttaattcag tgctacccca aaacagtgtt ttgggtggag
     2401 tcctcaaaac aaagcaacaa ttgaagacat taaatcattt tgatctgact aatggtgttc
     2461 tagttgagag cctaagtgaa gagccacttc cttctttgcg tcgaggccgg aaaagacact
     2521 gcaagaccaa gcacttagaa caaaatggct cccttaaaaa actgcgacaa accagtgggg
     2581 aggtgggtct ggccccaaca gacccagtcc tgcgagagat ggagcagaag cttcagcaag
     2641 aggaagaaga ccgacagttg gctctgcagt tgcagcgcat gttcgacaat gagaggcgga
     2701 ctgtgagccg gcgaaaagga agtgtggatc agtatctcct acggtccagc aacatggccg
     2761 gggccaagta gaacccagct ttcttgtaca aagtggctcg agcagggatc cgcccctctc
     2821 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     2881 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     2941 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     3001 gtctgttgaa tgtcgtgaag gaagcgtagg aattcgccag cacagtggtc gaccctgtgg
     3061 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3121 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3181 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3241 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3301 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3361 ggaggctttt ttggaggcct aggcttttgc aaaaagctta ccatgaccga gtacaagccc
     3421 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     3481 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     3541 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     3601 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     3661 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     3721 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     3781 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     3841 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     3901 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     3961 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg
     4021 cagcgcccga ccgaaaggag cgcacgaccc catgcatcga taaaataaaa gattttattt
     4081 agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa
     4141 gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa gttcagatca
     4201 aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
     4261 tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata
     4321 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
     4381 tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
     4441 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
     4501 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct
     4561 ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc
     4621 gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc
     4681 gggggtcttt cacatgcagc atgtatcaaa attaatttgg ttttttttct taagtattta
     4741 cattaaatgg ccatagttgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4801 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4861 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4921 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4981 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5041 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5101 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5161 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5221 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5281 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5341 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5401 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5461 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5521 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5581 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5641 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tgcggccgca
     5701 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5761 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5821 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5881 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5941 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6001 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6061 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6121 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6181 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6241 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6301 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6361 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6421 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6481 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6541 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6601 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6661 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6721 aagtgccacc tgcagcctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     6781 cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
     6841 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc
     6901 cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
     6961 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
     7021 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt
     7081 gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg
     7141 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt
     7201 ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca ccgacccacc
     7261 accgggaggc aagctggcca gcaacttatc tgtgtctgtc cgattgtcta gtgtctatga
     7321 ctgattttat gcgcctgcgt cggtactagt tagctaacta gctctgtatc tggcggaccc
     7381 gtggtggaac tgacgagttc tgaacacccg gccgcaaccc tgggagacgt cccagggact
     7441 ttgggggccg tttttgtggc ccgacctgag gaagggagtc gatgtggaat ccgaccccgt
     7501 caggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat
     7561 ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg
     7621 ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg
     7681 ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca
     7741 accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa
     7801 cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta
     7861 agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga
     7921 cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc
     7981 ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc
     8041 cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccggc cggatcccag
     8101 tgtggtggta ctaatagtaa tcaattacgg ggtcattagt tcatagccca tata
//
