LOCUS       sequence-342728-        6357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of isolated PARP1 CAT domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6357)
  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 6357)
  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-PARP1-CAT
FEATURES             Location/Qualifiers
     source          1..6357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     957..976
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(996..1073)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1279..1298
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1382..1401
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1382..1400
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1401..1425
                     /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             1440..1462
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1481..1498
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1508..1525
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2603..2620
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2673..2691)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2687..2734
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2771..3226
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2858..2877)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3068..3089
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3318..4133)
                     /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(3318..4133)
                     /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     4041..4060
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4125..4144
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4255..4843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4744..4763
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4997..5014
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5029..5168
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5094..5113)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5254..5276
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5270..5461)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6270..6357,1..995))
                     /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 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
       61 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
      121 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
      181 cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
      241 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
      301 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
      361 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
      421 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
      481 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
      541 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
      601 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
      661 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
      721 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
      781 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
      841 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
      901 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
      961 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     1021 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc
     1081 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
     1141 gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
     1201 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     1261 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     1321 gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa
     1381 ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt
     1441 ttgtttaact ttaagaagga gatataccat gggcagcagc catcatcatc atcatcacag
     1501 cagcggcctg gtgccgcgcg gcagccatat gaccaagtcc aagctcccca agccagttca
     1561 ggacctcatc aagatgatct ttgatgtgga aagtatgaag aaagccatgg tggagtatga
     1621 gatcgacctt cagaagatgc ccttggggaa gctgagcaaa aggcagatcc aggccgcata
     1681 ctccatcctc agtgaggtcc agcaggcggt gtctcagggc agcagcgact ctcagatcct
     1741 ggatctctca aatcgctttt acaccctgat cccccacgac tttgggatga agaagcctcc
     1801 gctcctgaac aatgcagaca gtgtgcaggc caaggcggaa atgcttgaca acctgctgga
     1861 catcgaggtg gcctacagtc tgctcagggg agggtctgat gatagcagca aggatcccat
     1921 cgatgtcaac tatgagaagc tcaaaactga cattaaggtg gttgacagag attctgaaga
     1981 agccgagatc atcaggaagt atgttaagaa cactcatgca accacacaca atgcgtatga
     2041 cttggaagtc atcgatatct ttaagataga gcgtgaaggc gaatgccagc gttacaagcc
     2101 ctttaagcag cttcataacc gaagattgct gtggcacggg tccaggacca ccaactttgc
     2161 tgggatcctg tcccagggtc ttcggatagc cccgcctgaa gcgcccgtga caggctacat
     2221 gtttggtaaa gggatctatt tcgctgacat ggtctccaag agtgccaact actgccatac
     2281 gtctcaggga gacccaatag gcttaatcct gttgggagaa gttgcccttg gaaacatgta
     2341 tgaactgaag cacgcttcac atatcagcaa gttacccaag ggcaagcaca gtgtcaaagg
     2401 tttgggcaaa actacccctg atccttcagc taacattagt ctggatggtg tagacgttcc
     2461 tcttgggacc gggatttcat ctggtgtgaa tgacacctct ctactatata acgagtacat
     2521 tgtctatgat attgctcagg taaatctgaa gtatctgctg aaactgaaat tcaattttaa
     2581 gacctccctg tggtaactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa
     2641 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     2701 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt
     2761 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2821 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2881 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2941 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3001 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3061 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3121 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3181 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     3241 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3301 cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc
     3361 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     3421 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     3481 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     3541 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
     3601 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     3661 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
     3721 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     3781 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg
     3841 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     3901 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     3961 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
     4021 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     4081 ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct cataacaccc
     4141 cttgtattac tgtttatgta agcagacagt tttattgttc atgaccaaaa tcccttaacg
     4201 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4261 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4321 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4381 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     4441 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4501 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4561 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4621 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4681 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4741 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4801 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4861 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4921 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4981 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     5041 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
     5101 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     5161 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     5221 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     5281 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     5341 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     5401 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     5461 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
     5521 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
     5581 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
     5641 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
     5701 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
     5761 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
     5821 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
     5881 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc
     5941 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     6001 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     6061 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     6121 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     6181 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     6241 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6301 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattg
//
