LOCUS       sequence-342731-        7857 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of PARP1 lacking Zn3 domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7857)
  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 7857)
  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-PARP1delZn3
FEATURES             Location/Qualifiers
     source          1..7857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     535..554
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(574..651)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     857..876
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     960..979
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        960..978
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    979..1003
                     /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             1018..1040
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1059..1076
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1086..1103
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             3681..3698
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3751..3769)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3765..3812
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3849..4304
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3936..3955)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4146..4167
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4396..5211)
                     /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(4396..5211)
                     /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     5119..5138
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5203..5222
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5333..5921
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5822..5841
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6075..6092
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6107..6246
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6172..6191)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6332..6354
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6348..6539)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7348..7857,1..573))
                     /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 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
       61 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
      121 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
      181 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
      241 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
      301 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
      361 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
      421 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
      481 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
      541 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
      601 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
      661 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
      721 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
      781 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
      841 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
      901 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
      961 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
     1021 gtttaacttt aagaaggaga tataccatgg gcagcagcca tcatcatcat catcacagca
     1081 gcggcctggt gccgcgcggc agccatatga tggcggagtc ttctgataag ctgtatcgag
     1141 tcgagtacgc caagagcggg cgcgcctctt gcaagaaatg cagcgagagc atccccaagg
     1201 actcgctccg gatggccatc atggtgcagt cgcccatgtt tgatggaaaa gtcccacact
     1261 ggtaccactt ctcctgcttc tggaaggtgg gccactccat ccggcaccct gacgttgagg
     1321 tggatgggtt ctctgagctt cggtgggatg accagcagaa agtcaagaag acagcggaag
     1381 ctggaggagt gacaggcaaa ggccaggatg gaattggtag caaggcagag aagactctgg
     1441 gtgactttgc agcagagtat gccaagtcca acagaagtac gtgcaagggg tgtatggaga
     1501 agatagaaaa gggccaggtg cgcctgtcca agaagatggt ggacccggag aagccacagc
     1561 taggcatgat tgaccgctgg taccatccag gctgctttgt caagaacagg gaggagctgg
     1621 gtttccggcc cgagtacagt gcgagtcagc tcaagggctt cagcctcctt gctacagagg
     1681 ataaggaagc cctgaagaag cagctcccag gagtcaagag tgaaggaaag agaaaaggcg
     1741 atgaggtggc ctcggctcct gctgctgtga actcctctgc ttcagcagat aagccattat
     1801 ccaacatgaa gatcctgact ctcgggaagc tgtcccggaa caaggatgaa gtgaaggcca
     1861 tgattgagaa actcgggggg aagttgacgg ggacggccaa caaggcttcc ctgtgcatca
     1921 gcaccaaaaa ggaggtggaa aagatgaata agaagatgga ggaagtaaag gaagccaaca
     1981 tccgagttgt gtctgaggac ttcctccagg acgtctccgc ctccaccaag agccttcagg
     2041 agttgttctt agcgcacatc ttgtcccctt ggggggcaga ggtgaaggca gagcctgttg
     2101 aagttgtggc cccaagaggg aagtcagggg ctgcgctctc caaaaaaagc aagggccagg
     2161 tcaaggagga aggtatcaac aaatctgaaa agagaatgaa attaactctt aaaggaggag
     2221 cagctgtgga tcctgattct ggactggaac actctgcgca tgtcctggag aaaggtggga
     2281 aggtcttcag tgccaccctt ggcctggtgg acatcgttaa aggaaccaac tcctactaca
     2341 agctccagct tctggaggac gacaaggaaa acaggtattg gatattcagg tcctggggcc
     2401 gtgtgggtac ggtgatcggt agcaacaaac tggaacagat gccttccaag gaggatgcca
     2461 ttgagcactt catgaaatta tatgaagaaa aaaccgggaa cgcttggcac tccaaaaatt
     2521 tcacgaagta tcccaaaaag ttctaccccc tggagattga ctatggccag gatgaagagg
     2581 cagtgaagaa gctgacagta aatcctggca ccaagtccaa gctccccaag ccagttcagg
     2641 acctcatcaa gatgatcttt gatgtggaaa gtatgaagaa agccatggtg gagtatgaga
     2701 tcgaccttca gaagatgccc ttggggaagc tgagcaaaag gcagatccag gccgcatact
     2761 ccatcctcag tgaggtccag caggcggtgt ctcagggcag cagcgactct cagatcctgg
     2821 atctctcaaa tcgcttttac accctgatcc cccacgactt tgggatgaag aagcctccgc
     2881 tcctgaacaa tgcagacagt gtgcaggcca aggcggaaat gcttgacaac ctgctggaca
     2941 tcgaggtggc ctacagtctg ctcaggggag ggtctgatga tagcagcaag gatcccatcg
     3001 atgtcaacta tgagaagctc aaaactgaca ttaaggtggt tgacagagat tctgaagaag
     3061 ccgagatcat caggaagtat gttaagaaca ctcatgcaac cacacacaat gcgtatgact
     3121 tggaagtcat cgatatcttt aagatagagc gtgaaggcga atgccagcgt tacaagccct
     3181 ttaagcagct tcataaccga agattgctgt ggcacgggtc caggaccacc aactttgctg
     3241 ggatcctgtc ccagggtctt cggatagccc cgcctgaagc gcccgtgaca ggctacatgt
     3301 ttggtaaagg gatctatttc gctgacatgg tctccaagag tgccaactac tgccatacgt
     3361 ctcagggaga cccaataggc ttaatcctgt tgggagaagt tgcccttgga aacatgtatg
     3421 aactgaagca cgcttcacat atcagcaagt tacccaaggg caagcacagt gtcaaaggtt
     3481 tgggcaaaac tacccctgat ccttcagcta acattagtct ggatggtgta gacgttcctc
     3541 ttgggaccgg gatttcatct ggtgtgaatg acacctctct actatataac gagtacattg
     3601 tctatgatat tgctcaggta aatctgaagt atctgctgaa actgaaattc aattttaaga
     3661 cctccctgtg gtaactcgag caccaccacc accaccactg agatccggct gctaacaaag
     3721 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     3781 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg
     3841 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3901 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3961 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4021 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4081 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4141 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4201 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4261 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     4321 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4381 ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     4441 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     4501 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     4561 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     4621 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     4681 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     4741 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     4801 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     4861 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     4921 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     4981 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     5041 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     5101 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     5161 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct
     5221 tgtattactg tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg
     5281 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5341 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5401 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5461 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5521 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5581 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5641 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5701 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5761 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5821 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5881 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5941 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6001 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     6061 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     6121 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
     6181 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     6241 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     6301 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     6361 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     6421 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     6481 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     6541 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
     6601 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
     6661 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
     6721 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
     6781 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
     6841 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
     6901 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
     6961 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg
     7021 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga
     7081 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag
     7141 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc
     7201 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
     7261 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     7321 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     7381 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     7441 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     7501 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     7561 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     7621 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     7681 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     7741 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     7801 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagac
//
