LOCUS       sequence-342747-        7911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of PARP1 lacking WGR domain but containing 
            L713F gain-of-function mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7911)
  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 7911)
  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-PARP1delWGR-L713F
FEATURES             Location/Qualifiers
     source          1..7911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(106..1188)
                     /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     1150..1169
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1189..1266)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1472..1491
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1575..1594
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1575..1593
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1594..1618
                     /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             1633..1655
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1674..1691
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1701..1718
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(4420..4438)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4434..4481
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4518..4973
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4605..4624)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4815..4836
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5065..5880)
                     /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(5065..5880)
                     /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     5788..5807
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5872..5891
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6002..6590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6491..6510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6744..6761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6776..6915
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6841..6860)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7001..7023
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7017..7208)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
       61 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
      121 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
      181 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
      241 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
      301 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
      361 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
      421 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
      481 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
      541 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
      601 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
      661 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
      721 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
      781 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
      841 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
      901 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
      961 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     1021 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     1081 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     1141 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     1201 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     1261 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     1321 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     1381 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     1441 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     1501 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
     1561 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
     1621 tctagaaata attttgttta actttaagaa ggagatatac catgggcagc agccatcatc
     1681 atcatcatca cagcagcggc ctggtgccgc gcggcagcca tatgatggcg gagtcttctg
     1741 ataagctgta tcgagtcgag tacgccaaga gcgggcgcgc ctcttgcaag aaatgcagcg
     1801 agagcatccc caaggactcg ctccggatgg ccatcatggt gcagtcgccc atgtttgatg
     1861 gaaaagtccc acactggtac cacttctcct gcttctggaa ggtgggccac tccatccggc
     1921 accctgacgt tgaggtggat gggttctctg agcttcggtg ggatgaccag cagaaagtca
     1981 agaagacagc ggaagctgga ggagtgacag gcaaaggcca ggatggaatt ggtagcaagg
     2041 cagagaagac tctgggtgac tttgcagcag agtatgccaa gtccaacaga agtacgtgca
     2101 aggggtgtat ggagaagata gaaaagggcc aggtgcgcct gtccaagaag atggtggacc
     2161 cggagaagcc acagctaggc atgattgacc gctggtacca tccaggctgc tttgtcaaga
     2221 acagggagga gctgggtttc cggcccgagt acagtgcgag tcagctcaag ggcttcagcc
     2281 tccttgctac agaggataag gaagccctga agaagcagct cccaggagtc aagagtgaag
     2341 gaaagagaaa aggcgatgag gtggatggag tggatgaagt ggcgaagaag aaatctaaaa
     2401 aagaaaaaga caaggatagt aagcttgaaa aagccctaaa ggctcagaac gacctgatct
     2461 ggaacatcaa ggacgagcta aagaaagtgt gttcaactaa tgacctgaag gagctactca
     2521 tcttcaacaa gcagcaagtg ccttctgggg agtcggcgat cttggaccga gtagctgatg
     2581 gcatggtgtt cggtgccctc cttccctgcg aggaatgctc gggtcagctg gtcttcaaga
     2641 gcgatgccta ttactgcact ggggacgtca ctgcctggac caagtgtatg gtcaagacac
     2701 agacacccaa ccggaaggag tgggtaaccc caaaggaatt ccgagaaatc tcttacctca
     2761 agaaattgaa ggttaaaaag caggaccgta tattcccccc agaaaccagc gcctccgtgg
     2821 cggccacgcc tccgccctcc acagcctcgg ctcctgctgc tgtgaactcc tctgcttcag
     2881 cagataagcc attatccaac atgaagatcc tgactctcgg gaagctgtcc cggaacaagg
     2941 atgaagtgaa ggccatgatt gagaaactcg gggggaagtt gacggggacg gccaacaagg
     3001 cttccctgtg catcagcacc aaaaaggagg tggaaaagat gaataagaag atggaggaag
     3061 taaaggaagc caacatccga gttgtgtctg aggacttcct ccaggacgtc tccgcctcca
     3121 ccaagagcct tcaggagttg ttcttagcgc acatcttgtc cccttggggg gcagaggtga
     3181 aggcagagcc tgttgaagtt gtggccccaa gagggaagtc aggggctgcg ctctccaaaa
     3241 aaagcaaggg ccaggtcaag gaggaaggta tcaacaaatc tgaaaccaag tccaagctcc
     3301 ccaagccagt tcaggacctc atcaagatga tctttgatgt ggaaagtatg aagaaagcca
     3361 tggtggagta tgagatcgac cttcagaaga tgcccttggg gaagctgagc aaaaggcaga
     3421 tccaggccgc atactccatc ttcagtgagg tccagcaggc ggtgtctcag ggcagcagcg
     3481 actctcagat cctggatctc tcaaatcgct tttacaccct gatcccccac gactttggga
     3541 tgaagaagcc tccgctcctg aacaatgcag acagtgtgca ggccaaggcg gaaatgcttg
     3601 acaacctgct ggacatcgag gtggcctaca gtctgctcag gggagggtct gatgatagca
     3661 gcaaggatcc catcgatgtc aactatgaga agctcaaaac tgacattaag gtggttgaca
     3721 gagattctga agaagccgag atcatcagga agtatgttaa gaacactcat gcaaccacac
     3781 acaatgcgta tgacttggaa gtcatcgata tctttaagat agagcgtgaa ggcgaatgcc
     3841 agcgttacaa gccctttaag cagcttcata accgaagatt gctgtggcac gggtccagga
     3901 ccaccaactt tgctgggatc ctgtcccagg gtcttcggat agccccgcct gaagcgcccg
     3961 tgacaggcta catgtttggt aaagggatct atttcgctga catggtctcc aagagtgcca
     4021 actactgcca tacgtctcag ggagacccaa taggcttaat cctgttggga gaagttgccc
     4081 ttggaaacat gtatgaactg aagcacgctt cacatatcag caagttaccc aagggcaagc
     4141 acagtgtcaa aggtttgggc aaaactaccc ctgatccttc agctaacatt agtctggatg
     4201 gtgtagacgt tcctcttggg accgggattt catctggtgt gaatgacacc tctctactat
     4261 ataacgagta cattgtctat gatattgctc aggtaaatct gaagtatctg ctgaaactga
     4321 aattcaattt taagacctcc ctgtggtaac tcgaccacca ccaccactga gatccggctg
     4381 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     4441 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     4501 ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4561 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4621 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4681 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4741 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4801 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4861 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4921 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     4981 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5041 atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa
     5101 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     5161 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     5221 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     5281 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc
     5341 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     5401 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
     5461 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     5521 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     5581 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     5641 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     5701 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     5761 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     5821 atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat
     5881 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg accaaaatcc
     5941 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6001 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6061 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6121 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     6181 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6241 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6301 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6361 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6421 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6481 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6541 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6601 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6661 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     6721 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     6781 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     6841 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     6901 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     6961 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     7021 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     7081 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     7141 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     7201 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     7261 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     7321 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     7381 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     7441 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     7501 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     7561 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     7621 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc
     7681 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     7741 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     7801 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     7861 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat g
//
