LOCUS       sequence-342733-        7545 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of PARP1 lacking Zn2 and BRCT domains.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7545)
  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 7545)
  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-PARP1delZn2delBRCT
FEATURES             Location/Qualifiers
     source          1..7545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     973..992
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1012..1089)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1295..1314
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1398..1417
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1398..1416
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1417..1441
                     /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             1456..1478
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1497..1514
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1524..1541
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             3807..3824
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3877..3895)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3891..3938
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3975..4430
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4062..4081)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4272..4293
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4522..5337)
                     /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(4522..5337)
                     /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     5245..5264
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5329..5348
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5459..6047
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5948..5967
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6201..6218
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6233..6372
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6298..6317)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6458..6480
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6474..6665)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7474..7545,1..1011))
                     /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 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
       61 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
      121 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
      181 ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
      241 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
      301 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
      361 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
      421 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
      481 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
      541 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
      601 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
      661 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
      721 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
      781 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
      841 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
      901 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
      961 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     1021 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     1081 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     1141 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     1201 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     1261 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     1321 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     1381 tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc
     1441 ccctctagaa ataattttgt ttaactttaa gaaggagata taccatgggc agcagccatc
     1501 atcatcatca tcacagcagc ggcctggtgc cgcgcggcag ccatatgatg gcggagtctt
     1561 ctgataagct gtatcgagtc gagtacgcca agagcgggcg cgcctcttgc aagaaatgca
     1621 gcgagagcat ccccaaggac tcgctccgga tggccatcat ggtgcagtcg cccatgtttg
     1681 atggaaaagt cccacactgg taccacttct cctgcttctg gaaggtgggc cactccatcc
     1741 ggcaccctga cgttgaggtg gatgggttct ctgagcttcg gtgggatgac cagcagaaag
     1801 tcaagaagac agcggaagct ggaggagtga caggcgatgg agtggatgaa gtggcgaaga
     1861 agaaatctaa aaaagaaaaa gacaaggata gtaagcttga aaaagcccta aaggctcaga
     1921 acgacctgat ctggaacatc aaggacgagc taaagaaagt gtgttcaact aatgacctga
     1981 aggagctact catcttcaac aagcagcaag tgccttctgg ggagtcggcg atcttggacc
     2041 gagtagctga tggcatggtg ttcggtgccc tccttccctg cgaggaatgc tcgggtcagc
     2101 tggtcttcaa gagcgatgcc tattactgca ctggggacgt cactgcctgg accaagtgta
     2161 tggtcaagac acagacaccc aaccggaagg agtgggtaac cccaaaggaa ttccgagaaa
     2221 tctcttacct caagaaattg aaggttaaaa agcaggaccg tatattcccc ccagaaacca
     2281 gcgcctccgt ggcggccacg cctccgccct ccacaaagag aatgaaatta actcttaaag
     2341 gaggagcagc tgtggatcct gattctggac tggaacactc tgcgcatgtc ctggagaaag
     2401 gtgggaaggt cttcagtgcc acccttggcc tggtggacat cgttaaagga accaactcct
     2461 actacaagct ccagcttctg gaggacgaca aggaaaacag gtattggata ttcaggtcct
     2521 ggggccgtgt gggtacggtg atcggtagca acaaactgga acagatgccg tccaaggagg
     2581 atgccattga gcacttcatg aaattatatg aagaaaaaac cgggaacgct tggcactcca
     2641 aaaatttcac gaagtatccc aaaaagttct accccctgga gattgactat ggccaggatg
     2701 aagaggcagt gaagaagctg acagtaaatc ctggcaccaa gtccaagctc cccaagccag
     2761 ttcaggacct catcaagatg atctttgatg tggaaagtat gaagaaagcc atggtggagt
     2821 atgagatcga ccttcagaag atgcccttgg ggaagctgag caaaaggcag atccaggccg
     2881 catactccat cctcagtgag gtccagcagg cggtgtctca gggcagcagc gactctcaga
     2941 tcctggatct ctcaaatcgc ttttacaccc tgatccccca cgactttggg atgaagaagc
     3001 ctccgctcct gaacaatgca gacagtgtgc aggccaaggc ggaaatgctt gacaacctgc
     3061 tggacatcga ggtggcctac agtctgctca ggggagggtc tgatgatagc agcaaggatc
     3121 ccatcgatgt caactatgag aagctcaaaa ctgacattaa ggtggttgac agagattctg
     3181 aagaagccga gatcatcagg aagtatgtta agaacactca tgcaaccaca cacaatgcgt
     3241 atgacttgga agtcatcgat atctttaaga tagagcgtga aggcgaatgc cagcgttaca
     3301 agccctttaa gcagcttcat aaccgaagat tgctgtggca cgggtccagg accaccaact
     3361 ttgctgggat cctgtcccag ggtcttcgga tagccccgcc tgaagcgccc gtgacaggct
     3421 acatgtttgg taaagggatc tatttcgctg acatggtctc caagagtgcc aactactgcc
     3481 atacgtctca gggagaccca ataggcttaa tcctgttggg agaagttgcc cttggaaaca
     3541 tgtatgaact gaagcacgct tcacatatca gcaagttacc caagggcaag cacagtgtca
     3601 aaggtttggg caaaactacc cctgatcctt cagctaacat tagtctggat ggtgtagacg
     3661 ttcctcttgg gaccgggatt tcatctggtg tgaatgacac ctctctacta tataacgagt
     3721 acattgtcta tgatattgct caggtaaatc tgaagtatct gctgaaactg aaattcaatt
     3781 ttaagacctc cctgtggtaa ctcgagcacc accaccacca ccactgagat ccggctgcta
     3841 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     3901 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
     3961 gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4021 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4081 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4141 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4201 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4261 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4321 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4381 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     4441 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4501 tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     4561 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     4621 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     4681 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     4741 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     4801 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     4861 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     4921 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     4981 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     5041 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     5101 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     5161 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     5221 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     5281 catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac
     5341 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt
     5401 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5461 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5521 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5581 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5641 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5701 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5761 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5821 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5881 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5941 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6001 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6061 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6121 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     6181 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     6241 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
     6301 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     6361 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     6421 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     6481 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     6541 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     6601 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     6661 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     6721 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     6781 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     6841 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     6901 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
     6961 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
     7021 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
     7081 cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg tggggccgcc
     7141 atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc agtgacgaag
     7201 gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat catcgtcgcg
     7261 ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac ctgtcctacg
     7321 agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc ccgcgcccac
     7381 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
     7441 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     7501 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggaga
//
