LOCUS       sequence-248735-        5943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG-HA-tagged RNF68 [Y252D] mutant in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5943)
  AUTHORS   Rona G, Roberti D, Yin Y, Pagan JK, Homer H, Sassani E, Zeke A, 
            Busino L, Rothenberg E, Pagano M
  TITLE     PARP1-dependent recruitment of the FBXL10-RNF68-RNF2 ubiquitin 
            ligase to sites of DNA damage controls H2A.Z loading.
  JOURNAL   Elife. 2018 Jul 9;7. pii: 38771. doi: 10.7554/eLife.38771.
  PUBMED    29985131
REFERENCE   2  (bases 1 to 5943)
  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"; journalName: "Elife."; 
            date: "2018-07-9"; volume: "7. pii: 38771. doi"; pages: " 
            10.7554/eLife.38771"
COMMENT     Sequence Label: FLAG-HA-RNF68 [Y252D]
FEATURES             Location/Qualifiers
     source          1..5943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..470
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    535..734
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             935..1351
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1281..1303
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1319..1335
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1380..1403
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1416..1442
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             1416..1442
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2266..2286)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2271..2600
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2451..2586
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2610..3209
                     /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(2610..2629)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3106..3126
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3430..3899
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4029..4046)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4200..4788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4280..4299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4948..5739,5740..5808))
                     /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     5571..5590
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5809..5913)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
       61 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      121 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      181 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      241 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
      301 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
      361 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
      421 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
      481 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggcaagctg
      541 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
      601 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      661 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg
      721 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct
      781 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
      841 gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg
      901 tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag
      961 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt
     1021 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg
     1081 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc
     1141 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc
     1201 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
     1261 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
     1321 ttatccagcc ctcactcctt ctctaggcgc cggccggatc ctccgctcga tgaaccatgg
     1381 actacaagga cgacgatgac aagctcgatg gaggataccc ctacgacgtg cccgactacg
     1441 ccgaaggaag atctgcgtct cctcaggggg gccagattgc gatcgcgatg aggcttcgga
     1501 accagctcca gtcagtgtac aagatggacc cgctacggaa cgaggaggag gttcgagtga
     1561 agatcaaaga cttgaatgaa cacattgttt gctgcctatg cgccggctac ttcgtggatg
     1621 ccaccaccat cacagagtgt cttcatactt tctgcaagag ttgtattgtg aagtacctcc
     1681 aaactagcaa gtactgcccc atgtgcaaca ttaagatcca cgagacacag ccactgctca
     1741 acctcaaact ggaccgggtc atgcaggaca tcgtgtataa gctggtgcct ggcttgcaag
     1801 acagtgaaga gaaacggatt cgggaattct accagtcccg aggtttggac cgggtcaccc
     1861 agcccactgg ggaagagcca gcactgagca acctcggcct ccccttcagc agctttgacc
     1921 actctaaagc ccactactat cgctatgatg agcagttgaa cctgtgcctg gagcggctga
     1981 gttctggcaa agacaagaat aaaagcgtcc tgcagaacaa gtatgtccga tgttctgtta
     2041 gagctgaggt acgccatctc cggagggtcc tgtgtcaccg cttgatgcta aaccctcagc
     2101 atgtgcagct cctttttgac aatgaagttc tccctgatca cgtgacaatg aagcagatat
     2161 ggctctcccg ctggttcggc aagccatccc ctttgctttt acaagacagt gtgaaagaga
     2221 agaggaggta gctcgagaat tcgccagcac agtggtcgac cctgtggaat gtgtgtcagt
     2281 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2341 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2401 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2461 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2521 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2581 gaggcctagg cttttgcaaa aagcttacca tgaccgagta caagcccacg gtgcgcctcg
     2641 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     2701 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     2761 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     2821 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     2881 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     2941 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     3001 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     3061 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     3121 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     3181 ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg
     3241 aaaggagcgc acgaccccat gcatcgataa aataaaagat tttatttagt ctccagaaaa
     3301 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt
     3361 gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag
     3421 atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     3481 cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca
     3541 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc
     3601 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
     3661 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
     3721 gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag
     3781 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
     3841 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac
     3901 atgcagcatg tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca
     3961 tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4021 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4081 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4141 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4201 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4261 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4321 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4381 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4441 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4501 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4561 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4621 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4681 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4741 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4801 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4861 atgagattat caaaaaggat cttcacctag atccttttgc ggccgcaaat caatctaaag
     4921 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4981 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5041 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     5101 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5161 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5221 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5281 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5341 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5401 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5461 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5521 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5581 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5641 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5701 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5761 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5821 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5881 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgc
     5941 agc
//
