LOCUS       sequence-342726-        7896 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of PARP1 lacking BRCT domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7896)
  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 7896)
  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-PARP1delBRCT
FEATURES             Location/Qualifiers
     source          1..7896
                     /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"
     CDS             4155..4172
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4225..4243)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4239..4286
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4323..4778
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4410..4429)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4620..4641
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4870..5685)
                     /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(4870..5685)
                     /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     5593..5612
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5677..5696
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5807..6395
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6296..6315
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6549..6566
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6581..6720
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6646..6665)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6806..6828
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6822..7013)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7822..7896,1..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 acaaagagaa tgaaattaac tcttaaagga ggagcagctg
     2701 tggatcctga ttctggactg gaacactctg cgcatgtcct ggagaaaggt gggaaggtct
     2761 tcagtgccac ccttggcctg gtggacatcg ttaaaggaac caactcctac tacaagctcc
     2821 agcttctgga ggacgacaag gaaaacaggt attggatatt caggtcctgg ggccgtgtgg
     2881 gtacggtgat cggtagcaac aaactggaac agatgccgtc caaggaggat gccattgagc
     2941 acttcatgaa attatatgaa gaaaaaaccg ggaacgcttg gcactccaaa aatttcacga
     3001 agtatcccaa aaagttctac cccctggaga ttgactatgg ccaggatgaa gaggcagtga
     3061 agaagctgac agtaaatcct ggcaccaagt ccaagctccc caagccagtt caggacctca
     3121 tcaagatgat ctttgatgtg gaaagtatga agaaagccat ggtggagtat gagatcgacc
     3181 ttcagaagat gcccttgggg aagctgagca aaaggcagat ccaggccgca tactccatcc
     3241 tcagtgaggt ccagcaggcg gtgtctcagg gcagcagcga ctctcagatc ctggatctct
     3301 caaatcgctt ttacaccctg atcccccacg actttgggat gaagaagcct ccgctcctga
     3361 acaatgcaga cagtgtgcag gccaaggcgg aaatgcttga caacctgctg gacatcgagg
     3421 tggcctacag tctgctcagg ggagggtctg atgatagcag caaggatccc atcgatgtca
     3481 actatgagaa gctcaaaact gacattaagg tggttgacag agattctgaa gaagccgaga
     3541 tcatcaggaa gtatgttaag aacactcatg caaccacaca caatgcgtat gacttggaag
     3601 tcatcgatat ctttaagata gagcgtgaag gcgaatgcca gcgttacaag ccctttaagc
     3661 agcttcataa ccgaagattg ctgtggcacg ggtccaggac caccaacttt gctgggatcc
     3721 tgtcccaggg tcttcggata gccccgcctg aagcgcccgt gacaggctac atgtttggta
     3781 aagggatcta tttcgctgac atggtctcca agagtgccaa ctactgccat acgtctcagg
     3841 gagacccaat aggcttaatc ctgttgggag aagttgccct tggaaacatg tatgaactga
     3901 agcacgcttc acatatcagc aagttaccca agggcaagca cagtgtcaaa ggtttgggca
     3961 aaactacccc tgatccttca gctaacatta gtctggatgg tgtagacgtt cctcttggga
     4021 ccgggatttc atctggtgtg aatgacacct ctctactata taacgagtac attgtctatg
     4081 atattgctca ggtaaatctg aagtatctgc tgaaactgaa attcaatttt aagacctccc
     4141 tgtggtaact cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa
     4201 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     4261 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg
     4321 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4381 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4441 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4501 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4561 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4621 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4681 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4741 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     4801 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4861 aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     4921 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     4981 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     5041 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     5101 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
     5161 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     5221 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     5281 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     5341 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     5401 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     5461 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     5521 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     5581 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     5641 taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     5701 actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgagtttt
     5761 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5821 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5881 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5941 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6001 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6061 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6121 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6181 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6241 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6301 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6361 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6421 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6481 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6541 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     6601 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga
     6661 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     6721 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     6781 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     6841 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     6901 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     6961 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     7021 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     7081 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     7141 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     7201 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     7261 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     7321 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     7381 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     7441 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat gccggcgata
     7501 atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg
     7561 gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag
     7621 cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata
     7681 aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg
     7741 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
     7801 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     7861 ctgcattaat gaatcggcca acgcgcgggg agaggc
//
