LOCUS Exported 5640 bp ds-DNA circular SYN 23-SEP-2021 DEFINITION Bacterial expression of isolated PARP1 Zn2 domain. ACCESSION . VERSION . KEYWORDS pET28_6xHis-PARP1-Zn2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5640) TITLE PARP and ALC1 plasmids REFERENCE 2 (bases 1 to 5640) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 23, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5640 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..20 /label=T7 /note="T7 promoter, forward primer" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 20..44 /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 59..81 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 100..117 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 127..144 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" primer_bind complement(575..593) /label=T7 Term /note="T7 terminator, reverse primer" terminator 589..636 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 673..1128 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(760..779) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 970..991 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(1220..2035) /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 1943..1962 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 2027..2046 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 2157..2745 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2646..2665 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2899..2916 /label=L4440 /note="L4440 vector, forward primer" misc_feature 2931..3070 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(2996..3015) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3156..3178 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3172..3363) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4172..5254) /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 5216..5235 /label=LacI-R /note="LacI, reverse primer" promoter complement(5255..5332) /gene="lacI" /label=lacI promoter primer_bind 5538..5557 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt 61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc 121 agcggcctgg tgccgcgcgg cagccatatg aaaggccagg atggaattgg tagcaaggca 181 gagaagactc tgggtgactt tgcagcagag tatgccaagt ccaacagaag tacgtgcaag 241 gggtgtatgg agaagataga aaagggccag gtgcgcctgt ccaagaagat ggtggacccg 301 gagaagccac agctaggcat gattgaccgc tggtaccatc caggctgctt tgtcaagaac 361 agggaggagc tgggtttccg gcccgagtac agtgcgagtc agctcaaggg cttcagcctc 421 cttgctacag aggataagga agccctgaag aagcagctcc caggagtcaa gagtgaagga 481 aagagaaaag gcgatgaggt gtaactcgag caccaccacc actgagatcc ggctgctaac 541 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc 601 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga 661 ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc 721 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt 781 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag 841 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt 901 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt 961 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt 1021 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt 1081 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt 1141 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 1201 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat 1261 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa 1321 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc 1381 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga 1441 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc 1501 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac 1561 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac 1621 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat 1681 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag 1741 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca 1801 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac 1861 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg 1921 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca 1981 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac 2041 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa 2101 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 2161 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 2221 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 2281 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag 2341 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 2401 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 2461 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 2521 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 2581 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 2641 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 2701 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 2761 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 2821 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 2881 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg 2941 tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa 3001 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt 3061 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct 3121 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt 3181 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg 3241 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag 3301 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt 3361 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg 3421 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg 3481 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca 3541 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc 3601 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga 3661 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca 3721 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg 3781 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat 3841 gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc 3901 ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct 3961 ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag 4021 ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg 4081 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg 4141 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct 4201 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 4261 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg 4321 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat 4381 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc 4441 ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc 4501 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca 4561 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg 4621 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga 4681 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct 4741 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt 4801 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat 4861 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt 4921 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg 4981 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg 5041 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca 5101 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg 5161 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat 5221 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt 5281 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct 5341 cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg 5401 ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg 5461 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga 5521 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg 5581 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat //