LOCUS       sequence-342181-        7911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of PARP1 lacking WGR domain.
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
FEATURES             Location/Qualifiers
     source          1..7911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     970..989
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1009..1086)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1292..1311
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1395..1414
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1395..1413
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1414..1438
                     /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             1453..1475
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1494..1511
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1521..1538
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(4240..4258)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4254..4301
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4338..4793
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4425..4444)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4635..4656
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4885..5700)
                     /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(4885..5700)
                     /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     5608..5627
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5692..5711
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5822..6410
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6311..6330
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6564..6581
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6596..6735
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6661..6680)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6821..6843
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6837..7028)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7837..7911,1..1008))
                     /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 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
       61 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
      121 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
      181 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
      241 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
      301 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
      361 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
      421 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
      481 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
      541 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
      601 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
      661 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
      721 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
      781 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
      841 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
      901 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
      961 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     1021 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     1081 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     1141 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     1201 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     1261 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     1321 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
     1381 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
     1441 tctagaaata attttgttta actttaagaa ggagatatac catgggcagc agccatcatc
     1501 atcatcatca cagcagcggc ctggtgccgc gcggcagcca tatgatggcg gagtcttctg
     1561 ataagctgta tcgagtcgag tacgccaaga gcgggcgcgc ctcttgcaag aaatgcagcg
     1621 agagcatccc caaggactcg ctccggatgg ccatcatggt gcagtcgccc atgtttgatg
     1681 gaaaagtccc acactggtac cacttctcct gcttctggaa ggtgggccac tccatccggc
     1741 accctgacgt tgaggtggat gggttctctg agcttcggtg ggatgaccag cagaaagtca
     1801 agaagacagc ggaagctgga ggagtgacag gcaaaggcca ggatggaatt ggtagcaagg
     1861 cagagaagac tctgggtgac tttgcagcag agtatgccaa gtccaacaga agtacgtgca
     1921 aggggtgtat ggagaagata gaaaagggcc aggtgcgcct gtccaagaag atggtggacc
     1981 cggagaagcc acagctaggc atgattgacc gctggtacca tccaggctgc tttgtcaaga
     2041 acagggagga gctgggtttc cggcccgagt acagtgcgag tcagctcaag ggcttcagcc
     2101 tccttgctac agaggataag gaagccctga agaagcagct cccaggagtc aagagtgaag
     2161 gaaagagaaa aggcgatgag gtggatggag tggatgaagt ggcgaagaag aaatctaaaa
     2221 aagaaaaaga caaggatagt aagcttgaaa aagccctaaa ggctcagaac gacctgatct
     2281 ggaacatcaa ggacgagcta aagaaagtgt gttcaactaa tgacctgaag gagctactca
     2341 tcttcaacaa gcagcaagtg ccttctgggg agtcggcgat cttggaccga gtagctgatg
     2401 gcatggtgtt cggtgccctc cttccctgcg aggaatgctc gggtcagctg gtcttcaaga
     2461 gcgatgccta ttactgcact ggggacgtca ctgcctggac caagtgtatg gtcaagacac
     2521 agacacccaa ccggaaggag tgggtaaccc caaaggaatt ccgagaaatc tcttacctca
     2581 agaaattgaa ggttaaaaag caggaccgta tattcccccc agaaaccagc gcctccgtgg
     2641 cggccacgcc tccgccctcc acagcctcgg ctcctgctgc tgtgaactcc tctgcttcag
     2701 cagataagcc attatccaac atgaagatcc tgactctcgg gaagctgtcc cggaacaagg
     2761 atgaagtgaa ggccatgatt gagaaactcg gggggaagtt gacggggacg gccaacaagg
     2821 cttccctgtg catcagcacc aaaaaggagg tggaaaagat gaataagaag atggaggaag
     2881 taaaggaagc caacatccga gttgtgtctg aggacttcct ccaggacgtc tccgcctcca
     2941 ccaagagcct tcaggagttg ttcttagcgc acatcttgtc cccttggggg gcagaggtga
     3001 aggcagagcc tgttgaagtt gtggccccaa gagggaagtc aggggctgcg ctctccaaaa
     3061 aaagcaaggg ccaggtcaag gaggaaggta tcaacaaatc tgaaaccaag tccaagctcc
     3121 ccaagccagt tcaggacctc atcaagatga tctttgatgt ggaaagtatg aagaaagcca
     3181 tggtggagta tgagatcgac cttcagaaga tgcccttggg gaagctgagc aaaaggcaga
     3241 tccaggccgc atactccatc ctcagtgagg tccagcaggc ggtgtctcag ggcagcagcg
     3301 actctcagat cctggatctc tcaaatcgct tttacaccct gatcccccac gactttggga
     3361 tgaagaagcc tccgctcctg aacaatgcag acagtgtgca ggccaaggcg gaaatgcttg
     3421 acaacctgct ggacatcgag gtggcctaca gtctgctcag gggagggtct gatgatagca
     3481 gcaaggatcc catcgatgtc aactatgaga agctcaaaac tgacattaag gtggttgaca
     3541 gagattctga agaagccgag atcatcagga agtatgttaa gaacactcat gcaaccacac
     3601 acaatgcgta tgacttggaa gtcatcgata tctttaagat agagcgtgaa ggcgaatgcc
     3661 agcgttacaa gccctttaag cagcttcata accgaagatt gctgtggcac gggtccagga
     3721 ccaccaactt tgctgggatc ctgtcccagg gtcttcggat agccccgcct gaagcgcccg
     3781 tgacaggcta catgtttggt aaagggatct atttcgctga catggtctcc aagagtgcca
     3841 actactgcca tacgtctcag ggagacccaa taggcttaat cctgttggga gaagttgccc
     3901 ttggaaacat gtatgaactg aagcacgctt cacatatcag caagttaccc aagggcaagc
     3961 acagtgtcaa aggtttgggc aaaactaccc ctgatccttc agctaacatt agtctggatg
     4021 gtgtagacgt tcctcttggg accgggattt catctggtgt gaatgacacc tctctactat
     4081 ataacgagta cattgtctat gatattgctc aggtaaatct gaagtatctg ctgaaactga
     4141 aattcaattt taagacctcc ctgtggtaac tcgaccacca ccaccactga gatccggctg
     4201 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     4261 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     4321 ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4381 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4441 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4501 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4561 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4621 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4681 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4741 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     4801 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4861 atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa
     4921 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     4981 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     5041 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     5101 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc
     5161 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     5221 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
     5281 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     5341 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     5401 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     5461 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     5521 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     5581 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     5641 atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat
     5701 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg accaaaatcc
     5761 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5821 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5881 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5941 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     6001 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6061 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6121 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6181 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6241 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6301 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6361 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6421 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6481 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     6541 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     6601 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     6661 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     6721 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     6781 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     6841 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     6901 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     6961 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     7021 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     7081 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     7141 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     7201 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     7261 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     7321 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     7381 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     7441 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc
     7501 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     7561 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     7621 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     7681 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     7741 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     7801 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     7861 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg c
//
