LOCUS       sequence-379245-        7711 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses PARP1 D770A 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_D770A-mCherry
FEATURES             Location/Qualifiers
     source          1..7711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      763..772
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             769..1479
                     /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(914..932)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1182..1201
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1602..1723
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1639..1658)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1693..1712
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1730..2185)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1867..1888)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2079..2098
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2212..2316
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2318..2675
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2318..2509
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2341..2361)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2526..2661
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2710..3504
                     /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             2710..3504
                     /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(2764..2783)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3374..3393
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3692..3711)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3736..3783
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4112..4700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4601..4620
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4875..5178
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5179..5382
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5333..5353
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 aacctgctgg ccatcgaggt ggcctacagt ctgctcaggg gagggtctga tgatagcagc
       61 aaggatccca tcgatgtcaa ctatgagaag ctcaaaactg acattaaggt ggttgacaga
      121 gattctgaag aagccgagat catcaggaag tatgttaaga acactcatgc aaccacacac
      181 agtgcgtatg acttggaagt catcgatatc tttaagatag agcgtgaagg cgaatgccag
      241 cgttacaagc cctttaagca gcttcataac cgaagattgc tgtggcacgg gtccaggacc
      301 accaactttg ctgggatcct gtcccagggt cttcggatag ccccgcctga agcgcccgtg
      361 acaggctaca tgtttggtaa agggatctat ttcgctgaca tggtctccaa gagtgccaac
      421 tactgccata cgtctcaggg agacccaata ggcttaatcc tgttgggaga agttgccctt
      481 ggaaacatgt atgaactgaa gcacgcttca catatcagca agttacccaa gggcaagcac
      541 agtgtcaaag gtttgggcaa aactacccct gatccttcag ctaacattag tctggatggt
      601 gtagacgttc ctcttgggac cgggatttca tctggtgtga atgacacctc tctactatat
      661 aacgagtaca ttgtctatga tattgctcag gtaaatctga agtatctgct gaaactgaaa
      721 ttcaatttta agacctccct gtggtcccgg gatccaccgg tcgccaccat ggtgagcaag
      781 ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag
      841 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
      901 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
      961 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
     1021 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     1081 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
     1141 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     1201 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc
     1261 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
     1321 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg
     1381 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     1441 cgccactcca ccggcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatca
     1501 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     1561 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     1621 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     1681 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     1741 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     1801 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     1861 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     1921 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     1981 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     2041 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     2101 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     2161 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     2221 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2281 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     2341 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2401 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     2461 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     2521 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2581 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     2641 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     2701 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     2761 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     2821 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     2881 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     2941 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3001 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3061 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3121 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3181 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     3241 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3301 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3361 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3421 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     3481 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     3541 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     3601 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     3661 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     3721 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     3781 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     3841 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     3901 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     3961 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4021 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     4081 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     4141 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4201 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     4261 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4321 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4381 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4441 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4501 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4561 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4621 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4681 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4741 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4801 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     4861 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     4921 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     4981 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     5041 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     5101 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     5161 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     5221 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     5281 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     5341 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
     5401 atccgctagc accatggcgg agtcttcgga taagctctat cgagtcgagt acgccaagag
     5461 cgggcgcgcc tcttgcaaga aatgcagcga gagcatcccc aaggactcgc tccggatggc
     5521 catcatggtg cagtcgccca tgtttgatgg aaaagtccca cactggtacc acttctcctg
     5581 cttctggaag gtgggccact ccatccggca ccctgacgtt gaggtggatg ggttctctga
     5641 gcttcggtgg gatgaccagc agaaagtcaa gaagacagcg gaagctggag gagtgacagg
     5701 caaaggccag gatggaattg gtagcaaggc agagaagact ctgggtgact ttgcagcaga
     5761 gtatgccaag tccaacagaa gtacgtgcaa ggggtgtatg gagaagatag aaaagggcca
     5821 ggtgcgcctg tccaagaaga tggtggaccc ggagaagcca cagctaggca tgattgaccg
     5881 ctggtaccat ccaggctgct ttgtcaagaa cagggaggag ctgggtttcc ggcccgagta
     5941 cagtgcgagt cagctcaagg gcttcagcct ccttgctaca gaggataagg aagccctgaa
     6001 gaagcagctc ccaggagtca agagtgaagg aaagagaaaa ggcgatgagg tggatggagt
     6061 ggatgaagtg gcgaagaaga aatctaaaaa agaaaaagac aaggatagta agcttgaaaa
     6121 agccctaaag gctcagaacg acctgatctg gaacatcaag gacgagctaa agaaagtgtg
     6181 ttcaactaat gacctgaagg agctactcat cttcaacaag cagcaagtgc cttctgggga
     6241 gtcggcgatc ttggaccgag tagctgatgg catggtgttc ggtgccctcc ttccctgcga
     6301 ggaatgctcg ggtcagctgg tcttcaagag cgatgcctat tactgcactg gggacgtcac
     6361 tgcctggacc aagtgtatgg tcaagacaca gacacccaac cggaaggagt gggtaacccc
     6421 aaaggaattc cgagaaatct cttacctcaa gaaattgaag gttaaaaagc aggaccgtat
     6481 attcccccca gaaaccagcg cctccgtggc ggccacgcct ccgccctcca cagcctcggc
     6541 tcctgctgct gtgaactcct ctgcttcagc agataagcca ttatccaaca tgaagatcct
     6601 gactctcggg aagctgtccc ggaacaagga tgaagtgaag gccatgattg agaaactcgg
     6661 ggggaagttg acggggacgg ccaacaaggc ttccctgtgc atcagcacca aaaaggaggt
     6721 ggaaaagatg aataagaaga tggaggaagt aaaggaagcc aacatccgag ttgtgtctga
     6781 ggacttcctc caggacgtct ccgcctccac caagagcctt caggagttgt tcttagcgca
     6841 catcttgtcc ccttgggggg cagaggtgaa ggcagagcct gttgaagttg tggccccaag
     6901 agggaagtca ggggctgcgc tctccaaaaa aagcaagggc caggtcaagg aggaaggtat
     6961 caacaaatct gaaaagagaa tgaaattaac tcttaaagga ggagcagctg tggatcctga
     7021 ttctggactg gaacactctg cgcatgtcct ggagaaaggt gggaaggtct tcagtgccac
     7081 ccttggcctg gtggacatcg ttaaaggaac caactcctac tacaagctcc agcttctgga
     7141 ggacgacaag gaaaacaggt attggatatt caggtcctgg ggccgtgtgg gtacggtgat
     7201 cggtagcaac aaactggaac agatgccgtc caaggaggat gccattgagc acttcatgaa
     7261 attatatgaa gaaaaaaccg ggaacgcttg gcactccaaa aatttcacga agtatcccaa
     7321 aaagttctac cccctggaga ttgactatgg ccaggatgaa gaggcagtga agaagctgac
     7381 agtaaatcct ggcaccaagt ccaagctccc caagccagtt caggacctca tcaagatgat
     7441 ctttgatgtg gaaagtatga agaaagccat ggtggagtat gagatcgacc ttcagaagat
     7501 gcccttgggg aagctgagca aaaggcagat ccaggccgca tactccatcc tcagtgaggt
     7561 ccagcaggcg gtgtctcagg gcagcagcga ctctcagatc ctggatctct caaatcgctt
     7621 ttacaccctg atcccccacg actttgggat gaagaagcct ccgctcctga acaatgcaga
     7681 cagtgtgcag gccaaggtgg aaatgcttga c
//
