LOCUS       sequence-345965-        7647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of an inactive PARP1 mutant lacking the ART 
            subdomain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7647)
  AUTHORS   Ooi SK, Sato S, Tomomori-Sato C, Zhang Y, Wen Z, Banks CAS, Washburn
            MP, Unruh JR, Florens L, Conaway RC, Conaway JW
  TITLE     Multiple roles for PARP1 in ALC1-dependent nucleosome remodeling.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Sep 7;118(36). pii: 2107277118. doi: 
            10.1073/pnas.2107277118.
  PUBMED    34465625
REFERENCE   2  (bases 1 to 7647)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2021-09-7"; volume: "118(36). pii: 
            2107277118. doi"; pages: " 10.1073/pnas.2107277118"
COMMENT     Sequence Label: pET28_6xHis-PARP1deltaART
FEATURES             Location/Qualifiers
     source          1..7647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2512..2529
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2582..2600)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2596..2643
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2680..3135
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2767..2786)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2977..2998
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3227..4042)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3227..4042)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3950..3969
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4034..4053
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4164..4752
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4653..4672
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4906..4923
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4938..5077
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5003..5022)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5163..5185
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5179..5370)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6179..7261)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     7223..7242
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7262..7339)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7545..7564
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgttcaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagccatatg atggcggagt cttctgataa gctgtatcga
      181 gtcgagtacg ccaagagcgg gcgcgcctct tgcaagaaat gcagcgagag catccccaag
      241 gactcgctcc ggatggccat catggtgcag tcgcccatgt ttgatggaaa agtcccacac
      301 tggtaccact tctcctgctt ctggaaggtg ggccactcca tccggcaccc tgacgttgag
      361 gtggatgggt tctctgagct tcggtgggat gaccagcaga aagtcaagaa gacagcggaa
      421 gctggaggag tgacaggcaa aggccaggat ggaattggta gcaaggcaga gaagactctg
      481 ggtgactttg cagcagagta tgccaagtcc aacagaagta cgtgcaaggg gtgtatggag
      541 aagatagaaa agggccaggt gcgcctgtcc aagaagatgg tggacccgga gaagccacag
      601 ctaggcatga ttgaccgctg gtaccatcca ggctgctttg tcaagaacag ggaggagctg
      661 ggtttccggc ccgagtacag tgcgagtcag ctcaagggct tcagcctcct tgctacagag
      721 gataaggaag ccctgaagaa gcagctccca ggagtcaaga gtgaaggaaa gagaaaaggc
      781 gatgaggtgg atggagtgga tgaagtggcg aagaagaaat ctaaaaaaga aaaagacaag
      841 gatagtaagc ttgaaaaagc cctaaaggct cagaacgacc tgatctggaa catcaaggac
      901 gagctaaaga aagtgtgttc aactaatgac ctgaaggagc tactcatctt caacaagcag
      961 caagtgcctt ctggggagtc ggcgatcttg gaccgagtag ctgatggcat ggtgttcggt
     1021 gccctccttc cctgcgagga atgctcgggt cagctggtct tcaagagcga tgcctattac
     1081 tgcactgggg acgtcactgc ctggaccaag tgtatggtca agacacagac acccaaccgg
     1141 aaggagtggg taaccccaaa ggaattccga gaaatctctt acctcaagaa attgaaggtt
     1201 aaaaagcagg accgtatatt ccccccagaa accagcgcct ccgtggcggc cacgcctccg
     1261 ccctccacag cctcggctcc tgctgctgtg aactcctctg cttcagcaga taagccatta
     1321 tccaacatga agatcctgac tctcgggaag ctgtcccgga acaaggatga agtgaaggcc
     1381 atgattgaga aactcggggg gaagttgacg gggacggcca acaaggcttc cctgtgcatc
     1441 agcaccaaaa aggaggtgga aaagatgaat aagaagatgg aggaagtaaa ggaagccaac
     1501 atccgagttg tgtctgagga cttcctccag gacgtctccg cctccaccaa gagccttcag
     1561 gagttgttct tagcgcacat cttgtcccct tggggggcag aggtgaaggc agagcctgtt
     1621 gaagttgtgg ccccaagagg gaagtcaggg gctgcgctct ccaaaaaaag caagggccag
     1681 gtcaaggagg aaggtatcaa caaatctgaa aagagaatga aattaactct taaaggagga
     1741 gcagctgtgg atcctgattc tggactggaa cactctgcgc atgtcctgga gaaaggtggg
     1801 aaggtcttca gtgccaccct tggcctggtg gacatcgtta aaggaaccaa ctcctactac
     1861 aagctccagc ttctggagga cgacaaggaa aacaggtatt ggatattcag gtcctggggc
     1921 cgtgtgggta cggtgatcgg tagcaacaaa ctggaacaga tgccgtccaa ggaggatgcc
     1981 attgagcact tcatgaaatt atatgaagaa aaaaccggga acgcttggca ctccaaaaat
     2041 ttcacgaagt atcccaaaaa gttctacccc ctggagattg actatggcca ggatgaagag
     2101 gcagtgaaga agctgacagt aaatcctggc accaagtcca agctccccaa gccagttcag
     2161 gacctcatca agatgatctt tgatgtggaa agtatgaaga aagccatggt ggagtatgag
     2221 atcgaccttc agaagatgcc cttggggaag ctgagcaaaa ggcagatcca ggccgcatac
     2281 tccatcctca gtgaggtcca gcaggcggtg tctcagggca gcagcgactc tcagatcctg
     2341 gatctctcaa atcgctttta caccctgatc ccccacgact ttgggatgaa gaagcctccg
     2401 ctcctgaaca atgcagacag tgtgcaggcc aaggcggaaa tgcttgacaa cctgctggac
     2461 atcgaggtgg cctacagtct gctcagggga gggtctgatg attaactcga gcaccaccac
     2521 caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     2581 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     2641 ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat
     2701 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2761 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2821 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2881 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2941 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3001 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3061 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3121 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3181 atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat
     3241 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     3301 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     3361 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     3421 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     3481 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     3541 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     3601 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     3661 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     3721 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     3781 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     3841 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     3901 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     3961 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg
     4021 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     4081 ttattgttca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4141 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4201 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4261 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     4321 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4381 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4441 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4501 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4561 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4621 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4681 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4741 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4801 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4861 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     4921 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4981 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     5041 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     5101 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     5161 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     5221 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     5281 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     5341 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     5401 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     5461 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     5521 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
     5581 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
     5641 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     5701 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     5761 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     5821 gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     5881 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     5941 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     6001 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     6061 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     6121 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     6181 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6241 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     6301 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     6361 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     6421 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc
     6481 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     6541 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     6601 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     6661 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     6721 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     6781 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     6841 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     6901 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     6961 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     7021 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     7081 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     7141 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     7201 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     7261 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     7321 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     7381 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     7441 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     7501 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     7561 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     7621 aggatcgaga tctcgatccc gcgaaat
//
