LOCUS       sequence-379244-        7711 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PARP1 R878A mutant in mammalian cells. Tagged with 
            mCherry..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7711)
  AUTHORS   Arnold MR, Langelier MF, Gartrell J, Kirby IT, Sanderson DJ, Bejan 
            DS, Sileikyte J, Sundalam SK, Nagarajan S, Marimuthu P, Duell AK, 
            Shelat AA, Pascal JM, Cohen MS
  TITLE     Allosteric regulation of DNA binding and target residence time drive
            the cytotoxicity of phthalazinone-based PARP-1 inhibitors.
  JOURNAL   Cell Chem Biol. 2022 Dec 15;29(12):1694-1708.e10. doi: 
            10.1016/j.chembiol.2022.11.006. Epub 2022 Dec 8.
  PUBMED    36493759
REFERENCE   2  (bases 1 to 7711)
  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.chembiol.2022.11.006"; journalName: "Cell Chem Biol"; 
            date: "2022-12-15- 15"; volume: "29"; issue: "12"; pages: 
            "1694-1708.e10"
COMMENT     Sequence Label: pCMV-PARP1_R878A-mCherry
FEATURES             Location/Qualifiers
     source          1..7711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      600..609
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             606..1316
                     /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(751..769)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1019..1038
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1439..1560
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1476..1495)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1530..1549
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1567..2022)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1704..1725)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1916..1935
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2049..2153
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2155..2512
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2155..2346
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2178..2198)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2363..2498
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2547..3341
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2547..3341
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2601..2620)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3211..3230
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3529..3548)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3573..3620
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3949..4537
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4438..4457
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4712..5015
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5016..5219
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5170..5190
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ctcatgcaac cacacacaat gcgtatgact tggaagtcat cgatatcttt aagatagagc
       61 gtgaaggcga atgccagcgt tacaagccct ttaagcagct tcataaccga agattgctgt
      121 ggcacgggtc caggaccacc aactttgctg ggatcctgtc ccagggtctt gccatagccc
      181 cgcctgaagc gcccgtgaca ggctacatgt ttggtaaagg gatctatttc gctgacatgg
      241 tctccaagag tgccaactac taccatacgt ctcagggaga cccaataggc ttaatcctgt
      301 tgggagaagt tgcccttgga aacatgtatg aactgaagca cgcttcacat atcagcaggt
      361 tacccaaggg caagcacagt gtcaaaggtt tgggcaaaac tacccctgat ccttcagcta
      421 acattagtct ggatggtgta gacgttcctc ttgggaccgg gatttcatct ggtgtgatag
      481 acacctctct actatataac gagtacattg tctatgatat tgctcaggta aatctgaagt
      541 atctgctgaa actgaaattc aattttaaga cctccctgtg gtcccgggat ccaccggtcg
      601 ccaccatggt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
      661 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
      721 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
      781 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
      841 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
      901 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
      961 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
     1021 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
     1081 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
     1141 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     1201 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     1261 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaaagcg
     1321 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     1381 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     1441 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     1501 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     1561 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     1621 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     1681 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     1741 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     1801 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     1861 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     1921 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     1981 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     2041 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2101 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     2161 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2221 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     2281 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     2341 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     2401 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     2461 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     2521 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     2581 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     2641 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     2701 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     2761 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     2821 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     2881 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     2941 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3001 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3061 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3121 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3181 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3241 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3301 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     3361 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     3421 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     3481 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     3541 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     3601 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     3661 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     3721 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     3781 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     3841 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     3901 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3961 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4021 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4081 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4141 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4201 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4261 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4321 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4381 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4441 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4501 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4561 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4621 ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt
     4681 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     4741 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     4801 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     4861 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     4921 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     4981 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     5041 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     5101 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     5161 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     5221 gtttagtgaa ccgtcagatc cgctagcacc atggcggagt cttcggataa gctctatcga
     5281 gtcgagtacg ccaagagcgg gcgcgcctct tgcaagaaat gcagcgagag catccccaag
     5341 gactcgctcc ggatggccat catggtgcag tcgcccatgt ttgatggaaa agtcccacac
     5401 tggtaccact tctcctgctt ctggaaggtg ggccactcca tccggcaccc tgacgttgag
     5461 gtggatgggt tctctgagct tcggtgggat gaccagcaga aagtcaagaa gacagcggaa
     5521 gctggaggag tgacaggcaa aggccaggat ggaattggta gcaaggcaga gaagactctg
     5581 ggtgactttg cagcagagta tgccaagtcc aacagaagta cgtgcaaggg gtgtatggag
     5641 aagatagaaa agggccaggt gcgcctgtcc aagaagatgg tggacccgga gaagccacag
     5701 ctaggcatga ttgaccgctg gtaccatcca ggctgctttg tcaagaacag ggaggagctg
     5761 ggtttccggc ccgagtacag tgcgagtcag ctcaagggct tcagcctcct tgctacagag
     5821 gataaggaag ccctgaagaa gcagctccca ggagtcaaga gtgaaggaaa gagaaaaggc
     5881 gatgaggtgg atggagtgga tgaagtggcg aagaagaaat ctaaaaaaga aaaagacaag
     5941 gatagtaagc ttgaaaaagc cctaaaggct cagaacgacc tgatctggaa catcaaggac
     6001 gagctaaaga aagtgtgttc aactaatgac ctgaaggagc tactcatctt caacaagcag
     6061 caagtgcctt ctggggagtc ggcgatcttg gaccgagtag ctgatggcat ggtgttcggt
     6121 gccctccttc cctgcgagga atgctcgggt cagctggtct tcaagagcga tgcctattac
     6181 tgcactgggg acgtcactgc ctggaccaag tgtatggtca agacacagac acccaaccgg
     6241 aaggagtggg taaccccaaa ggaattccga gaaatctctt acctcaagaa attgaaggtt
     6301 aaaaagcagg accgtatatt ccccccagaa accagcgcct ccgtggcggc cacgcctccg
     6361 ccctccacag cctcggctcc tgctgctgtg aactcctctg cttcagcaga taagccatta
     6421 tccaacatga agatcctgac tctcgggaag ctgtcccgga acaaggatga agtgaaggcc
     6481 atgattgaga aactcggggg gaagttgacg gggacggcca acaaggcttc cctgtgcatc
     6541 agcaccaaaa aggaggtgga aaagatgaat aagaagatgg aggaagtaaa ggaagccaac
     6601 atccgagttg tgtctgagga cttcctccag gacgtctccg cctccaccaa gagccttcag
     6661 gagttgttct tagcgcacat cttgtcccct tggggggcag aggtgaaggc agagcctgtt
     6721 gaagttgtgg ccccaagagg gaagtcaggg gctgcgctct ccaaaaaaag caagggccag
     6781 gtcaaggagg aaggtatcaa caaatctgaa aagagaatga aattaactct taaaggagga
     6841 gcagctgtgg atcctgattc tggactggaa cactctgcgc atgtcctgga gaaaggtggg
     6901 aaggtcttca gtgccaccct tggcctggtg gacatcgtta aaggaaccaa ctcctactac
     6961 aagctccagc ttctggagga cgacaaggaa aacaggtatt ggatattcag gtcctggggc
     7021 cgtgtgggta cggtgatcgg tagcaacaaa ctggaacaga tgccgtccaa ggaggatgcc
     7081 attgagcact tcatgaaatt atatgaagaa aaaaccggga acgcttggca ctccaaaaat
     7141 ttcacgaagt atcccaaaaa gttctacccc ctggagattg actatggcca ggatgaagag
     7201 gcagtgaaga agctgacagt aaatcctggc accaagtcca agctccccaa gccagttcag
     7261 gacctcatca agatgatctt tgatgtggaa agtatgaaga aagccatggt ggagtatgag
     7321 atcgaccttc agaagatgcc cttggggaag ctgagcaaaa ggcagatcca ggccgcatac
     7381 tccatcctca gtgaggtcca gcaggcggtg tctcagggca gcagcgactc tcagatcctg
     7441 gatctctcaa atcgctttta caccctgatc ccccacgact ttgggatgaa gaagcctccg
     7501 ctcctgaaca atgcagacag tgtgcaggcc aaggcggaaa tgcttgacaa cctgctggac
     7561 atcgaggtgg cctacagtct gctcagggga gggtctgatg atagcagcaa ggatcccatc
     7621 gatgtcaact atgagaagct caaaactgac attaaggtgg ttgacagaga ttctgaagaa
     7681 gccgagatca tcaggaagta tgttaagaac a
//
