LOCUS       sequence-342183-        7986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of PARP1 lacking Zn2 domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7986)
  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 7986)
  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-PARP1delZn2
FEATURES             Location/Qualifiers
     source          1..7986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1..23
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             42..59
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             69..86
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2793..2810
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2863..2881)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2877..2924
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2961..3416
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3048..3067)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3258..3279
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3508..4323)
                     /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(3508..4323)
                     /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     4231..4250
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4315..4334
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4445..5033
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4934..4953
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5187..5204
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5219..5358
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5284..5303)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5444..5466
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5460..5651)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6460..7542)
                     /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     7504..7523
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7543..7620)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7826..7845
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     7929..7948
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7929..7947
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7948..7972
                     /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)."
ORIGIN
        1 tttgtttaac tttaagaagg agatatacca tgggcagcag ccatcatcat catcatcaca
       61 gcagcggcct ggtgccgcgc ggcagccata tgatggcgga gtcttctgat aagctgtatc
      121 gagtcgagta cgccaagagc gggcgcgcct cttgcaagaa atgcagcgag agcatcccca
      181 aggactcgct ccggatggcc atcatggtgc agtcgcccat gtttgatgga aaagtcccac
      241 actggtacca cttctcctgc ttctggaagg tgggccactc catccggcac cctgacgttg
      301 aggtggatgg gttctctgag cttcggtggg atgaccagca gaaagtcaag aagacagcgg
      361 aagctggagg agtgacaggc gatggagtgg atgaagtggc gaagaagaaa tctaaaaaag
      421 aaaaagacaa ggatagtaag cttgaaaaag ccctaaaggc tcagaacgac ctgatctgga
      481 acatcaagga cgagctaaag aaagtgtgtt caactaatga cctgaaggag ctactcatct
      541 tcaacaagca gcaagtgcct tctggggagt cggcgatctt ggaccgagta gctgatggca
      601 tggtgttcgg tgccctcctt ccctgcgagg aatgctcggg tcagctggtc ttcaagagcg
      661 atgcctatta ctgcactggg gacgtcactg cctggaccaa gtgtatggtc aagacacaga
      721 cacccaaccg gaaggagtgg gtaaccccaa aggaattccg agaaatctct tacctcaaga
      781 aattgaaggt taaaaagcag gaccgtatat tccccccaga aaccagcgcc tccgtggcgg
      841 ccacgcctcc gccctccaca gcctcggctc ctgctgctgt gaactcctct gcttcagcag
      901 ataagccatt atccaacatg aagatcctga ctctcgggaa gctgtcccgg aacaaggatg
      961 aagtgaaggc catgattgag aaactcgggg ggaagttgac ggggacggcc aacaaggctt
     1021 ccctgtgcat cagcaccaaa aaggaggtgg aaaagatgaa taagaagatg gaggaagtaa
     1081 aggaagccaa catccgagtt gtgtctgagg acttcctcca ggacgtctcc gcctccacca
     1141 agagccttca ggagttgttc ttagcgcaca tcttgtcccc ttggggggca gaggtgaagg
     1201 cagagcctgt tgaagttgtg gccccaagag ggaagtcagg ggctgcgctc tccaaaaaaa
     1261 gcaagggcca ggtcaaggag gaaggtatca acaaatctga aaagagaatg aaattaactc
     1321 ttaaaggagg agcagctgtg gatcctgatt ctggactgga acactctgcg catgtcctgg
     1381 agaaaggtgg gaaggtcttc agtgccaccc ttggcctggt ggacatcgtt aaaggaacca
     1441 actcctacta caagctccag cttctggagg acgacaagga aaacaggtat tggatattca
     1501 ggtcctgggg ccgtgtgggt acggtgatcg gtagcaacaa actggaacag atgccgtcca
     1561 aggaggatgc cattgagcac ttcatgaaat tatatgaaga aaaaaccggg aacgcttggc
     1621 actccaaaaa tttcacgaag tatcccaaaa agttctaccc cctggagatt gactatggcc
     1681 aggatgaaga ggcagtgaag aagctgacag taaatcctgg caccaagtcc aagctcccca
     1741 agccagttca ggacctcatc aagatgatct ttgatgtgga aagtatgaag aaagccatgg
     1801 tggagtatga gatcgacctt cagaagatgc ccttggggaa gctgagcaaa aggcagatcc
     1861 aggccgcata ctccatcctc agtgaggtcc agcaggcggt gtctcagggc agcagcgact
     1921 ctcagatcct ggatctctca aatcgctttt acaccctgat cccccacgac tttgggatga
     1981 agaagcctcc gctcctgaac aatgcagaca gtgtgcaggc caaggcggaa atgcttgaca
     2041 acctgctgga catcgaggtg gcctacagtc tgctcagggg agggtctgat gatagcagca
     2101 aggatcccat cgatgtcaac tatgagaagc tcaaaactga cattaaggtg gttgacagag
     2161 attctgaaga agccgagatc atcaggaagt atgttaagaa cactcatgca accacacaca
     2221 atgcgtatga cttggaagtc atcgatatct ttaagataga gcgtgaaggc gaatgccagc
     2281 gttacaagcc ctttaagcag cttcataacc gaagattgct gtggcacggg tccaggacca
     2341 ccaactttgc tgggatcctg tcccagggtc ttcggatagc cccgcctgaa gcgcccgtga
     2401 caggctacat gtttggtaaa gggatctatt tcgctgacat ggtctccaag agtgccaact
     2461 actgccatac gtctcaggga gacccaatag gcttaatcct gttgggagaa gttgcccttg
     2521 gaaacatgta tgaactgaag cacgcttcac atatcagcaa gttacccaag ggcaagcaca
     2581 gtgtcaaagg tttgggcaaa actacccctg atccttcagc taacattagt ctggatggtg
     2641 tagacgttcc tcttgggacc gggatttcat ctggtgtgaa tgacacctct ctactatata
     2701 acgagtacat tgtctatgat attgctcagg taaatctgaa gtatctgctg aaactgaaat
     2761 tcaattttaa gacctccctg tggtaactcg agcaccacca ccaccaccac tgagatccgg
     2821 ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
     2881 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     2941 tatccggatt ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3001 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3061 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3121 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3181 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3241 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3301 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3361 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     3421 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3481 aatatgtatc cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg
     3541 caatttattc atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga
     3601 aggagaaaac tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat
     3661 tccgactcgt ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc
     3721 aagtgagaaa tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat
     3781 ttctttccag acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc
     3841 aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt
     3901 aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc
     3961 aacaatattt tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg
     4021 gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg
     4081 aagaggcata aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc
     4141 aacgctacct ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg
     4201 atagattgtc gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc
     4261 agcatccatg ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct
     4321 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgaccaaaa
     4381 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4441 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4501 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4561 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     4621 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4681 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4741 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4801 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4861 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4921 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4981 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5041 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5101 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     5161 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     5221 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct
     5281 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     5341 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     5401 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     5461 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     5521 tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     5581 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     5641 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     5701 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     5761 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     5821 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     5881 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     5941 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     6001 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     6061 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggg
     6121 gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg
     6181 acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc
     6241 gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt
     6301 cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc
     6361 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt
     6421 gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6481 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6541 cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc
     6601 cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag
     6661 gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc
     6721 gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg
     6781 cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc
     6841 attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc
     6901 cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg
     6961 cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac
     7021 cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg
     7081 tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc
     7141 aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag
     7201 aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac
     7261 cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc
     7321 gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg
     7381 ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg
     7441 cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac
     7501 accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg
     7561 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
     7621 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
     7681 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
     7741 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     7801 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     7861 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
     7921 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga
     7981 aataat
//
