LOCUS       sequence-211938-        7146 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses full length PARP1 with K to Q mutations in 
            automodification domain in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7146)
  AUTHORS   Liszczak G, Diehl KL, Dann GP, Muir TW
  TITLE     Acetylation blocks DNA damage-induced chromatin ADP-ribosylation.
  JOURNAL   Nat Chem Biol. 2018 Jul 16. pii: 10.1038/s41589-018-0097-1. doi: 
            10.1038/s41589-018-0097-1.
  PUBMED    30013063
REFERENCE   2  (bases 1 to 7146)
  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: "Nat Chem 
            Biol. 2018 Jul 16. pii: 10.1038/s41589-018-0097-1. doi: 
            10.1038/s41589-018-0097-1."
COMMENT     Sequence Label: pCMV-PARP1-3xFlag-7xKQ
FEATURES             Location/Qualifiers
     source          1..7146
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        175..478
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        479..682
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     633..653
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             774..797
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             807..830
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             840..863
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    4048..4169
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4085..4104)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4139..4158
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4176..4631)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4313..4334)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4525..4544
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4658..4762
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4764..5121
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4764..4955
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4787..4807)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4972..5107
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5156..5950
                     /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             5156..5950
                     /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(5210..5229)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5820..5839
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6138..6157)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6182..6229
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6558..7146
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7047..7066
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
      121 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      181 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      241 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      301 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      361 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      421 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      481 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      541 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      601 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      661 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac
      721 tcagatctcg agctcaagct tcgaattctg cagtcgacgg taccatgggt tcagactata
      781 aagatgatga tgataaaggt ggctccgact acaaggatga cgatgacaag ggaggtagtg
      841 actacaaaga cgatgacgac aagggcggat ctggcggatc tatggcggag tcttcggata
      901 agctctatcg agtcgagtac gccaagagcg ggcgcgcctc ttgcaagaaa tgcagcgaga
      961 gcatccccaa ggactcgctc cggatggcca tcatggtgca gtcgcccatg tttgatggaa
     1021 aagtcccaca ctggtaccac ttctcctgct tctggaaggt gggccactcc atccggcacc
     1081 ctgacgttga ggtggatggg ttctctgagc ttcggtggga tgatcagcag aaagtcaaga
     1141 agacagcgga agctggagga gtgacaggca aaggccagga tggaattggt agcaaggcag
     1201 agaagactct gggtgacttt gcagcagagt atgccaagtc caacagaagt acgtgcaagg
     1261 ggtgtatgga gaagatagaa aagggccagg tgcgcctgtc caagaagatg gtggacccgg
     1321 agaagccaca gctaggcatg attgaccgct ggtaccatcc aggctgcttt gtcaagaaca
     1381 gggaggagct gggtttccgg cccgagtaca gtgcgagtca gctcaagggc ttcagcctcc
     1441 ttgctacaga ggataaagaa gccctgaaga agcagctccc aggagtcaag agtgaaggaa
     1501 agagaaaagg cgatgaggtg gatggagtgg atgaagtggc gaagaagaaa tctaaaaaag
     1561 aaaaagacaa ggatagtaag cttgaaaaag ccctaaaggc tcagaacgac ctgatctgga
     1621 acatcaagga cgagctaaag aaagtgtgtt caactaatga cctgaaggag ctactcatct
     1681 tcaacaagca gcaagtgcct tctggggagt cggcgatctt ggaccgagta gccgatggca
     1741 tggtgttcgg tgccctcctt ccctgcgagg aatgctcggg tcagctggtc ttcaagagcg
     1801 atgcctatta ctgcactggg gacgtcactg cctggaccaa gtgtatggtc aagacacaga
     1861 cacccaaccg gaaggagtgg gtaaccccaa aggaattccg agaaatctct tacctcaaga
     1921 aattgaaggt taaaaaacag gaccgtatat tccccccaga aaccagcgcc tccgtggcgg
     1981 ccacgcctcc gccctccaca gcctcggctc ctgctgctgt gaactcctct gcttcagcag
     2041 ataagccatt atccaacatg aagatcctga ctctcgggaa gctgtcccgg aacaaggatg
     2101 aagtgaaggc catgattgag aaactcgggg ggaagttgac ggggacggcc aacaaggctt
     2161 ccctgtgcat cagcaccaaa aaggaggtgg aaaagatgaa taagaagatg gaggaagtaa
     2221 aggaagccaa catccgagtt gtgtctgagg acttcctcca ggacgtctcc gcctccacca
     2281 agagccttca ggagttgttc ttagcgcaca tcttgtcccc ttggggggca gaggtgaagg
     2341 cagagcctgt tgaagttgtg gccccaagag ggcagtcagg ggctgcgctc tcccagcaga
     2401 gccagggcca ggtcaaggag gaaggtatca accagtctga acagagaatg cagttaactc
     2461 ttaaaggagg agcagctgtg gatcctgatt ctggactgga acactctgcg catgtcctgg
     2521 agaaaggtgg gaaggtcttc agtgccaccc ttggcctggt ggacatcgtt aaaggaacca
     2581 actcctacta caagctgcag cttctggagg acgacaagga aaacaggtat tggatattca
     2641 ggtcctgggg ccgtgtgggt acggtgatcg gtagcaacaa actggaacag atgccgtcca
     2701 aggaggatgc cattgagcac ttcatgaaat tatatgaaga aaaaaccggg aacgcttggc
     2761 actccaaaaa tttcacgaag tatcccaaaa agttctaccc cctggagatt gactatggcc
     2821 aggatgaaga ggcagtgaag aagctgacag taaatcctgg caccaagtcc aagctcccca
     2881 agccagttca ggacctcatc aagatgatct ttgatgtgga aagtatgaag aaagccatgg
     2941 tggagtatga gatcgacctt cagaagatgc ccttggggaa gctgagcaaa aggcagatcc
     3001 aggccgcata ctccatcctc agtgaggtcc agcaggcggt gtctcagggc agcagcgact
     3061 ctcagatcct ggatctctca aatcgctttt acaccctgat cccccacgac tttgggatga
     3121 agaagcctcc gctcctgaac aatgcagaca gtgtgcaggc caaggcggaa atgcttgaca
     3181 acctgctgga catcgaggtg gcctacagtc tgctcagggg agggtctgat gatagcagca
     3241 aggatcccat cgatgtcaac tatgagaagc tcaaaactga cattaaggtg gttgacagag
     3301 attctgaaga agccgagatc atcaggaagt atgttaagaa cactcatgca accacacaca
     3361 atgcgtatga cttggaagtc atcgatatct ttaagataga gcgtgaaggc gaatgccagc
     3421 gttacaagcc ctttaagcag cttcataacc gaagattgct gtggcacggg tccaggacca
     3481 ccaactttgc tgggatcctg tcccagggtc ttcggatagc cccgcctgaa gcgcccgtga
     3541 caggctacat gtttggtaaa gggatctatt tcgctgacat ggtctccaag agtgccaact
     3601 actgccatac gtctcaggga gacccaatag gcttaatcct gttgggagaa gttgcccttg
     3661 gaaacatgta tgaactgaag cacgcttcac atatcagcaa gttacccaag ggcaagcaca
     3721 gtgtcaaagg tttgggcaaa actacccctg atccttcagc taacattagt ctggatggtg
     3781 tagacgttcc tcttgggacc gggatttcat ctggtgtgaa tgacacctct ctactatata
     3841 acgagtacat tgtctatgat attgctcagg taaatctgaa gtatctgctg aaactgaaat
     3901 tcaattttaa gacctccctg tggtgagcgg ccgcgactct agatcataat cagccatacc
     3961 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     4021 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     4081 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4141 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     4201 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     4261 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     4321 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     4381 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     4441 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     4501 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     4561 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     4621 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4681 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4741 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     4801 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4861 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4921 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4981 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5041 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     5101 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     5161 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     5221 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     5281 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     5341 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     5401 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     5461 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     5521 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     5581 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     5641 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     5701 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     5761 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5821 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5881 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5941 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     6001 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     6061 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     6121 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     6181 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     6241 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     6301 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     6361 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     6421 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6481 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6541 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6601 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6661 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6721 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6781 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6841 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6901 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6961 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7021 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7081 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7141 tggaaa
//
