LOCUS Exported 7896 bp ds-DNA circular SYN 15-SEP-2021 DEFINITION Bacterial expression of PARP1 lacking BRCT domain. ACCESSION . VERSION . KEYWORDS pET28_6xHis-PARP1delBRCT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7896) TITLE HOLD-Conaway Lab Plasmids REFERENCE 2 (bases 1 to 7896) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 15, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7896 /organism="synthetic DNA construct" /mol_type="other DNA" 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 4155..4172 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(4225..4243) /label=T7 Term /note="T7 terminator, reverse primer" terminator 4239..4286 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 4323..4778 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4410..4429) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4620..4641 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(4870..5685) /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 5593..5612 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 5677..5696 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 5807..6395 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6296..6315 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6549..6566 /label=L4440 /note="L4440 vector, forward primer" misc_feature 6581..6720 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(6646..6665) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6806..6828 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(6822..7013) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(7822..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 tatgatggcg gagtcttctg 1561 ataagctgta tcgagtcgag tacgccaaga gcgggcgcgc ctcttgcaag aaatgcagcg 1621 agagcatccc caaggactcg ctccggatgg ccatcatggt gcagtcgccc atgtttgatg 1681 gaaaagtccc acactggtac cacttctcct gcttctggaa ggtgggccac tccatccggc 1741 accctgacgt tgaggtggat gggttctctg agcttcggtg ggatgaccag cagaaagtca 1801 agaagacagc ggaagctgga ggagtgacag gcaaaggcca ggatggaatt ggtagcaagg 1861 cagagaagac tctgggtgac tttgcagcag agtatgccaa gtccaacaga agtacgtgca 1921 aggggtgtat ggagaagata gaaaagggcc aggtgcgcct gtccaagaag atggtggacc 1981 cggagaagcc acagctaggc atgattgacc gctggtacca tccaggctgc tttgtcaaga 2041 acagggagga gctgggtttc cggcccgagt acagtgcgag tcagctcaag ggcttcagcc 2101 tccttgctac agaggataag gaagccctga agaagcagct cccaggagtc aagagtgaag 2161 gaaagagaaa aggcgatgag gtggatggag tggatgaagt ggcgaagaag aaatctaaaa 2221 aagaaaaaga caaggatagt aagcttgaaa aagccctaaa ggctcagaac gacctgatct 2281 ggaacatcaa ggacgagcta aagaaagtgt gttcaactaa tgacctgaag gagctactca 2341 tcttcaacaa gcagcaagtg ccttctgggg agtcggcgat cttggaccga gtagctgatg 2401 gcatggtgtt cggtgccctc cttccctgcg aggaatgctc gggtcagctg gtcttcaaga 2461 gcgatgccta ttactgcact ggggacgtca ctgcctggac caagtgtatg gtcaagacac 2521 agacacccaa ccggaaggag tgggtaaccc caaaggaatt ccgagaaatc tcttacctca 2581 agaaattgaa ggttaaaaag caggaccgta tattcccccc agaaaccagc gcctccgtgg 2641 cggccacgcc tccgccctcc acaaagagaa tgaaattaac tcttaaagga ggagcagctg 2701 tggatcctga ttctggactg gaacactctg cgcatgtcct ggagaaaggt gggaaggtct 2761 tcagtgccac ccttggcctg gtggacatcg ttaaaggaac caactcctac tacaagctcc 2821 agcttctgga ggacgacaag gaaaacaggt attggatatt caggtcctgg ggccgtgtgg 2881 gtacggtgat cggtagcaac aaactggaac agatgccgtc caaggaggat gccattgagc 2941 acttcatgaa attatatgaa gaaaaaaccg ggaacgcttg gcactccaaa aatttcacga 3001 agtatcccaa aaagttctac cccctggaga ttgactatgg ccaggatgaa gaggcagtga 3061 agaagctgac agtaaatcct ggcaccaagt ccaagctccc caagccagtt caggacctca 3121 tcaagatgat ctttgatgtg gaaagtatga agaaagccat ggtggagtat gagatcgacc 3181 ttcagaagat gcccttgggg aagctgagca aaaggcagat ccaggccgca tactccatcc 3241 tcagtgaggt ccagcaggcg gtgtctcagg gcagcagcga ctctcagatc ctggatctct 3301 caaatcgctt ttacaccctg atcccccacg actttgggat gaagaagcct ccgctcctga 3361 acaatgcaga cagtgtgcag gccaaggcgg aaatgcttga caacctgctg gacatcgagg 3421 tggcctacag tctgctcagg ggagggtctg atgatagcag caaggatccc atcgatgtca 3481 actatgagaa gctcaaaact gacattaagg tggttgacag agattctgaa gaagccgaga 3541 tcatcaggaa gtatgttaag aacactcatg caaccacaca caatgcgtat gacttggaag 3601 tcatcgatat ctttaagata gagcgtgaag gcgaatgcca gcgttacaag ccctttaagc 3661 agcttcataa ccgaagattg ctgtggcacg ggtccaggac caccaacttt gctgggatcc 3721 tgtcccaggg tcttcggata gccccgcctg aagcgcccgt gacaggctac atgtttggta 3781 aagggatcta tttcgctgac atggtctcca agagtgccaa ctactgccat acgtctcagg 3841 gagacccaat aggcttaatc ctgttgggag aagttgccct tggaaacatg tatgaactga 3901 agcacgcttc acatatcagc aagttaccca agggcaagca cagtgtcaaa ggtttgggca 3961 aaactacccc tgatccttca gctaacatta gtctggatgg tgtagacgtt cctcttggga 4021 ccgggatttc atctggtgtg aatgacacct ctctactata taacgagtac attgtctatg 4081 atattgctca ggtaaatctg aagtatctgc tgaaactgaa attcaatttt aagacctccc 4141 tgtggtaact cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa 4201 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct 4261 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg 4321 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 4381 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 4441 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt 4501 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg 4561 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag 4621 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt 4681 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt 4741 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat 4801 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg 4861 aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg 4921 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag 4981 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc 5041 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg 5101 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc 5161 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat 5221 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac 5281 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga 5341 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa 5401 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt 5461 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg 5521 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga 5581 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt 5641 taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt 5701 actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgagtttt 5761 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt 5821 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 5881 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga 5941 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag 6001 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata 6061 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 6121 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga 6181 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 6241 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 6301 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 6361 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 6421 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt 6481 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga 6541 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc 6601 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga 6661 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg 6721 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc 6781 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca 6841 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca 6901 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc 6961 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc 7021 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg 7081 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa 7141 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc 7201 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc 7261 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa 7321 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac 7381 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc 7441 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat gccggcgata 7501 atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg 7561 gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag 7621 cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata 7681 aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg 7741 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa 7801 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 7861 ctgcattaat gaatcggcca acgcgcgggg agaggc //