LOCUS       sequence-342182-        8049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of PARP1 lacking Zn1 domain.
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
FEATURES             Location/Qualifiers
     source          1..8049
                     /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             4308..4325
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4378..4396)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4392..4439
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4476..4931
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4563..4582)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4773..4794
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5023..5838)
                     /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(5023..5838)
                     /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     5746..5765
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5830..5849
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5960..6548
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6449..6468
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6702..6719
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6734..6873
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6799..6818)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6959..6981
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6975..7166)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7975..8049,1..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 tatgaaaggc caggatggaa
     1561 ttggtagcaa ggcagagaag actctgggtg actttgcagc agagtatgcc aagtccaaca
     1621 gaagtacgtg caaggggtgt atggagaaga tagaaaaggg ccaggtgcgc ctgtccaaga
     1681 agatggtgga cccggagaag ccacagctag gcatgattga ccgctggtac catccaggct
     1741 gctttgtcaa gaacagggag gagctgggtt tccggcccga gtacagtgcg agtcagctca
     1801 agggcttcag cctccttgct acagaggata aggaagccct gaagaagcag ctcccaggag
     1861 tcaagagtga aggaaagaga aaaggcgatg aggtggatgg agtggatgaa gtggcgaaga
     1921 agaaatctaa aaaagaaaaa gacaaggata gtaagcttga aaaagcccta aaggctcaga
     1981 acgacctgat ctggaacatc aaggacgagc taaagaaagt gtgttcaact aatgacctga
     2041 aggagctact catcttcaac aagcagcaag tgccttctgg ggagtcggcg atcttggacc
     2101 gagtagctga tggcatggtg ttcggtgccc tccttccctg cgaggaatgc tcgggtcagc
     2161 tggtcttcaa gagcgatgcc tattactgca ctggggacgt cactgcctgg accaagtgta
     2221 tggtcaagac acagacaccc aaccggaagg agtgggtaac cccaaaggaa ttccgagaaa
     2281 tctcttacct caagaaattg aaggttaaaa agcaggaccg tatattcccc ccagaaacca
     2341 gcgcctccgt ggcggccacg cctccgccct ccacagcctc ggctcctgct gctgtgaact
     2401 cctctgcttc agcagataag ccattatcca acatgaagat cctgactctc gggaagctgt
     2461 cccggaacaa ggatgaagtg aaggccatga ttgagaaact cggggggaag ttgacgggga
     2521 cggccaacaa ggcttccctg tgcatcagca ccaaaaagga ggtggaaaag atgaataaga
     2581 agatggagga agtaaaggaa gccaacatcc gagttgtgtc tgaggacttc ctccaggacg
     2641 tctccgcctc caccaagagc cttcaggagt tgttcttagc gcacatcttg tccccttggg
     2701 gggcagaggt gaaggcagag cctgttgaag ttgtggcccc aagagggaag tcaggggctg
     2761 cgctctccaa aaaaagcaag ggccaggtca aggaggaagg tatcaacaaa tctgaaaaga
     2821 gaatgaaatt aactcttaaa ggaggagcag ctgtggatcc tgattctgga ctggaacact
     2881 ctgcgcatgt cctggagaaa ggtgggaagg tcttcagtgc cacccttggc ctggtggaca
     2941 tcgttaaagg aaccaactcc tactacaagc tccagcttct ggaggacgac aaggaaaaca
     3001 ggtattggat attcaggtcc tggggccgtg tgggtacggt gatcggtagc aacaaactgg
     3061 aacagatgcc gtccaaggag gatgccattg agcacttcat gaaattatat gaagaaaaaa
     3121 ccgggaacgc ttggcactcc aaaaatttca cgaagtatcc caaaaagttc taccccctgg
     3181 agattgacta tggccaggat gaagaggcag tgaagaagct gacagtaaat cctggcacca
     3241 agtccaagct ccccaagcca gttcaggacc tcatcaagat gatctttgat gtggaaagta
     3301 tgaagaaagc catggtggag tatgagatcg accttcagaa gatgcccttg gggaagctga
     3361 gcaaaaggca gatccaggcc gcatactcca tcctcagtga ggtccagcag gcggtgtctc
     3421 agggcagcag cgactctcag atcctggatc tctcaaatcg cttttacacc ctgatccccc
     3481 acgactttgg gatgaagaag cctccgctcc tgaacaatgc agacagtgtg caggccaagg
     3541 cggaaatgct tgacaacctg ctggacatcg aggtggccta cagtctgctc aggggagggt
     3601 ctgatgatag cagcaaggat cccatcgatg tcaactatga gaagctcaaa actgacatta
     3661 aggtggttga cagagattct gaagaagccg agatcatcag gaagtatgtt aagaacactc
     3721 atgcaaccac acacaatgcg tatgacttgg aagtcatcga tatctttaag atagagcgtg
     3781 aaggcgaatg ccagcgttac aagcccttta agcagcttca taaccgaaga ttgctgtggc
     3841 acgggtccag gaccaccaac tttgctggga tcctgtccca gggtcttcgg atagccccgc
     3901 ctgaagcgcc cgtgacaggc tacatgtttg gtaaagggat ctatttcgct gacatggtct
     3961 ccaagagtgc caactactgc catacgtctc agggagaccc aataggctta atcctgttgg
     4021 gagaagttgc ccttggaaac atgtatgaac tgaagcacgc ttcacatatc agcaagttac
     4081 ccaagggcaa gcacagtgtc aaaggtttgg gcaaaactac ccctgatcct tcagctaaca
     4141 ttagtctgga tggtgtagac gttcctcttg ggaccgggat ttcatctggt gtgaatgaca
     4201 cctctctact atataacgag tacattgtct atgatattgc tcaggtaaat ctgaagtatc
     4261 tgctgaaact gaaattcaat tttaagacct ccctgtggta actcgagcac caccaccacc
     4321 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     4381 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     4441 tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag
     4501 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4561 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4621 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4681 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4741 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4801 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4861 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4921 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4981 ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag
     5041 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     5101 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     5161 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     5221 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     5281 caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     5341 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     5401 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     5461 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     5521 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     5581 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     5641 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     5701 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     5761 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc
     5821 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     5881 tgttcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5941 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6001 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6061 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     6121 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6181 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6241 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6301 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6361 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6421 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6481 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6541 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6601 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     6661 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     6721 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     6781 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     6841 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     6901 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     6961 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     7021 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     7081 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     7141 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     7201 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     7261 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     7321 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     7381 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     7441 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     7501 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     7561 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     7621 atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     7681 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     7741 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     7801 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     7861 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     7921 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     7981 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     8041 gggagaggc
//
