LOCUS       sequence-342577-        8049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of PARP1 lacking Zn1 domain but containing 
            L713F gain-of-function mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8049)
  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 8049)
  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-PARP1delZn1-L713F
FEATURES             Location/Qualifiers
     source          1..8049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     434..453
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(473..550)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     756..775
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     859..878
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        859..877
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    878..902
                     /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)."
     RBS             917..939
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             958..975
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             985..1002
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             3772..3789
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3842..3860)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3856..3903
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3940..4395
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4027..4046)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4237..4258
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4487..5302)
                     /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(4487..5302)
                     /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     5210..5229
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5294..5313
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5424..6012
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5913..5932
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6166..6183
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6198..6337
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6263..6282)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6423..6445
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6439..6630)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7439..8049,1..472))
                     /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"
ORIGIN
        1 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
       61 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
      121 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
      181 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
      241 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
      301 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca
      361 ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct
      421 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
      481 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt
      541 cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc
      601 agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg
      661 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc
      721 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
      781 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag
      841 atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt
      901 cccctctaga aataattttg tttaacttta agaaggagat ataccatggg cagcagccat
      961 catcatcatc atcacagcag cggcctggtg ccgcgcggca gccatatgaa aggccaggat
     1021 ggaattggta gcaaggcaga gaagactctg ggtgactttg cagcagagta tgccaagtcc
     1081 aacagaagta cgtgcaaggg gtgtatggag aagatagaaa agggccaggt gcgcctgtcc
     1141 aagaagatgg tggacccgga gaagccacag ctaggcatga ttgaccgctg gtaccatcca
     1201 ggctgctttg tcaagaacag ggaggagctg ggtttccggc ccgagtacag tgcgagtcag
     1261 ctcaagggct tcagcctcct tgctacagag gataaggaag ccctgaagaa gcagctccca
     1321 ggagtcaaga gtgaaggaaa gagaaaaggc gatgaggtgg atggagtgga tgaagtggcg
     1381 aagaagaaat ctaaaaaaga aaaagacaag gatagtaagc ttgaaaaagc cctaaaggct
     1441 cagaacgacc tgatctggaa catcaaggac gagctaaaga aagtgtgttc aactaatgac
     1501 ctgaaggagc tactcatctt caacaagcag caagtgcctt ctggggagtc ggcgatcttg
     1561 gaccgagtag ctgatggcat ggtgttcggt gccctccttc cctgcgagga atgctcgggt
     1621 cagctggtct tcaagagcga tgcctattac tgcactgggg acgtcactgc ctggaccaag
     1681 tgtatggtca agacacagac acccaaccgg aaggagtggg taaccccaaa ggaattccga
     1741 gaaatctctt acctcaagaa attgaaggtt aaaaagcagg accgtatatt ccccccagaa
     1801 accagcgcct ccgtggcggc cacgcctccg ccctccacag cctcggctcc tgctgctgtg
     1861 aactcctctg cttcagcaga taagccatta tccaacatga agatcctgac tctcgggaag
     1921 ctgtcccgga acaaggatga agtgaaggcc atgattgaga aactcggggg gaagttgacg
     1981 gggacggcca acaaggcttc cctgtgcatc agcaccaaaa aggaggtgga aaagatgaat
     2041 aagaagatgg aggaagtaaa ggaagccaac atccgagttg tgtctgagga cttcctccag
     2101 gacgtctccg cctccaccaa gagccttcag gagttgttct tagcgcacat cttgtcccct
     2161 tggggggcag aggtgaaggc agagcctgtt gaagttgtgg ccccaagagg gaagtcaggg
     2221 gctgcgctct ccaaaaaaag caagggccag gtcaaggagg aaggtatcaa caaatctgaa
     2281 aagagaatga aattaactct taaaggagga gcagctgtgg atcctgattc tggactggaa
     2341 cactctgcgc atgtcctgga gaaaggtggg aaggtcttca gtgccaccct tggcctggtg
     2401 gacatcgtta aaggaaccaa ctcctactac aagctccagc ttctggagga cgacaaggaa
     2461 aacaggtatt ggatattcag gtcctggggc cgtgtgggta cggtgatcgg tagcaacaaa
     2521 ctggaacaga tgccgtccaa ggaggatgcc attgagcact tcatgaaatt atatgaagaa
     2581 aaaaccggga acgcttggca ctccaaaaat ttcacgaagt atcccaaaaa gttctacccc
     2641 ctggagattg actatggcca ggatgaagag gcagtgaaga agctgacagt aaatcctggc
     2701 accaagtcca agctccccaa gccagttcag gacctcatca agatgatctt tgatgtggaa
     2761 agtatgaaga aagccatggt ggagtatgag atcgaccttc agaagatgcc cttggggaag
     2821 ctgagcaaaa ggcagatcca ggccgcatac tccatcttca gtgaggtcca gcaggcggtg
     2881 tctcagggca gcagcgactc tcagatcctg gatctctcaa atcgctttta caccctgatc
     2941 ccccacgact ttgggatgaa gaagcctccg ctcctgaaca atgcagacag tgtgcaggcc
     3001 aaggcggaaa tgcttgacaa cctgctggac atcgaggtgg cctacagtct gctcagggga
     3061 gggtctgatg atagcagcaa ggatcccatc gatgtcaact atgagaagct caaaactgac
     3121 attaaggtgg ttgacagaga ttctgaagaa gccgagatca tcaggaagta tgttaagaac
     3181 actcatgcaa ccacacacaa tgcgtatgac ttggaagtca tcgatatctt taagatagag
     3241 cgtgaaggcg aatgccagcg ttacaagccc tttaagcagc ttcataaccg aagattgctg
     3301 tggcacgggt ccaggaccac caactttgct gggatcctgt cccagggtct tcggatagcc
     3361 ccgcctgaag cgcccgtgac aggctacatg tttggtaaag ggatctattt cgctgacatg
     3421 gtctccaaga gtgccaacta ctgccatacg tctcagggag acccaatagg cttaatcctg
     3481 ttgggagaag ttgcccttgg aaacatgtat gaactgaagc acgcttcaca tatcagcaag
     3541 ttacccaagg gcaagcacag tgtcaaaggt ttgggcaaaa ctacccctga tccttcagct
     3601 aacattagtc tggatggtgt agacgttcct cttgggaccg ggatttcatc tggtgtgaat
     3661 gacacctctc tactatataa cgagtacatt gtctatgata ttgctcaggt aaatctgaag
     3721 tatctgctga aactgaaatt caattttaag acctccctgt ggtaactcga gcaccaccac
     3781 caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     3841 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     3901 ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat
     3961 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4021 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4081 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4141 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4201 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4261 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4321 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4381 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4441 atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat
     4501 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     4561 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     4621 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     4681 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     4741 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     4801 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     4861 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     4921 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     4981 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     5041 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     5101 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     5161 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     5221 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg
     5281 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     5341 ttattgttca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5401 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5461 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5521 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     5581 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5641 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5701 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5761 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5821 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5881 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5941 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6001 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6061 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6121 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6181 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     6241 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     6301 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     6361 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     6421 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     6481 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     6541 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     6601 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
     6661 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
     6721 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
     6781 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
     6841 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
     6901 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
     6961 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
     7021 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
     7081 gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     7141 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     7201 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     7261 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     7321 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     7381 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     7441 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     7501 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     7561 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     7621 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     7681 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc
     7741 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     7801 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     7861 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     7921 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     7981 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     8041 aaataatac
//
