LOCUS       sequence-248724-        8358 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry-tagged RNF68 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8358)
  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 8358)
  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: mCherry-RNF68
FEATURES             Location/Qualifiers
     source          1..8358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             585..760
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    823..1022
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1223..1639
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1569..1591
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1607..1623
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1714..2017
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2018..2221
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      2260..2269
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2266..2973
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2411..2429)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2679..2698
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     promoter        3825..4324
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3860..3883)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4257..4276
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4345..4944
                     /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(4345..4364)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4841..4861
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5129..5561
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5664..5686)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5822..5842)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5827..6156
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6007..6142
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6178..6197
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6278..6295)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6449..7037)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6529..6548)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7208..7999,8000..8068))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7831..7850
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8069..8173)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8241..8259
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcaataa aagagcccac
      601 aacccctcac tcggcgcgcc agtcttccga tagactgcgt cgcccgggta cccgtattcc
      661 caataaagcc tcttgctgtt tgcatccgaa tcgtggtctc gctgttcctt gggagggtct
      721 cctctgagtg attgactacc cacgacgggg gtctttcatt tgggggctcg tccgggattt
      781 ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc cagcaactta
      841 tctgtgtctg tccgattgtc tagtgtctat gtttgatgtt atgcgcctgc gtctgtacta
      901 gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc
      961 cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg
     1021 aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga
     1081 gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga accgaagccg
     1141 cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
     1201 tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg
     1261 tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg
     1321 ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg
     1381 cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca
     1441 tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc
     1501 tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc
     1561 gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct
     1621 cactccttct ctaggcgccg gaattgaaga tcatagttat taatagtaat caattacggg
     1681 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     1741 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     1801 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     1861 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     1921 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     1981 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     2041 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     2101 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     2161 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     2221 tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt gagcaagggc
     2281 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     2341 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     2401 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     2461 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     2521 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     2581 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     2641 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     2701 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     2761 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     2821 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     2881 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     2941 cactccaccg gcggcatgga cgagctgtac aagtccggac tcagatctgc gtctcctcag
     3001 gggggccaga ttgcgatcgc gatgaggctt cggaaccagc tccagtcagt gtacaagatg
     3061 gacccgctac ggaacgagga ggaggttcga gtgaagatca aagacttgaa tgaacacatt
     3121 gtttgctgcc tatgcgccgg ctacttcgtg gatgccacca ccatcacaga gtgtcttcat
     3181 actttctgca agagttgtat tgtgaagtac ctccaaacta gcaagtactg ccccatgtgc
     3241 aacattaaga tccacgagac acagccactg ctcaacctca aactggaccg ggtcatgcag
     3301 gacatcgtgt ataagctggt gcctggcttg caagacagtg aagagaaacg gattcgggaa
     3361 ttctaccagt cccgaggttt ggaccgggtc acccagccca ctggggaaga gccagcactg
     3421 agcaacctcg gcctcccctt cagcagcttt gaccactcta aagcccacta ctatcgctat
     3481 gatgagcagt tgaacctgtg cctggagcgg ctgagttctg gcaaagacaa gaataaaagc
     3541 gtcctgcaga acaagtatgt ccgatgttct gttagagctg aggtacgcca tctccggagg
     3601 gtcctgtgtc accgcttgat gctaaaccct cagcatgtgc agctcctttt tgacaatgaa
     3661 gttctccctg atcacatgac aatgaagcag atatggctct cccgctggtt cggcaagcca
     3721 tcccctttgc ttttacaata cagtgtgaaa gagaagagga ggtagctcga cggtaccgcg
     3781 ggcccgggat ccaccggatc tagataactg atcataattc taccgggtag gggaggcgct
     3841 tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac
     3901 aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt
     3961 ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc
     4021 cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt
     4081 gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata
     4141 gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg
     4201 gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt
     4261 ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct
     4321 ttcgacctgc agcccaagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc
     4381 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     4441 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     4501 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     4561 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     4621 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     4681 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     4741 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     4801 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     4861 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     4921 atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg
     4981 agcgcacgac cccatgcatc ggcgtctcga tcgagatatc agtggtccag gctctagttt
     5041 tgactcaaca atatcaccag ctgaagccta tagagtacga gccatagata aaataaaaga
     5101 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
     5161 tactagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga
     5221 agttcagatc aaggtcagga acagatggaa cagggtcgat cgaccctaga gaaccatcag
     5281 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     5341 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     5401 caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat
     5461 ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct
     5521 cctctgagtg attgactacc cgtcagcggg ggtctttcat ttgggggctc gtccgggatc
     5581 gggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg ctgcctcgcg
     5641 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     5701 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     5761 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta gatccggctg
     5821 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5881 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5941 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     6001 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     6061 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     6121 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttactggctt aactatgcgg
     6181 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     6241 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6301 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6361 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6421 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6481 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6541 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6601 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6661 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6721 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6781 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6841 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     6901 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6961 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7021 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7081 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7141 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7201 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7261 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7321 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7381 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7441 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7501 tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7561 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7621 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7681 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7741 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7801 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
     7861 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7921 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7981 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8041 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8101 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8161 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     8221 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattag
     8281 cttggccatt gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc
     8341 caacattacc gccatgtt
//
