LOCUS Exported 5715 bp ds-DNA circular SYN 23-SEP-2021 DEFINITION Bacterial expression of isolated PARP1 WGR domain. ACCESSION . VERSION . KEYWORDS pET28_6xHis-PARP1-WGR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5715) TITLE PARP and ALC1 plasmids REFERENCE 2 (bases 1 to 5715) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 23, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5715 /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" CDS 580..597 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(650..668) /label=T7 Term /note="T7 terminator, reverse primer" terminator 664..711 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 748..1203 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(835..854) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1045..1066 /label=F1ori-F /note="F1 origin, forward primer" CDS complement(1295..2110) /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 2018..2037 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 2102..2121 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin 2232..2820 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2721..2740 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2974..2991 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3006..3145 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3071..3090) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3231..3253 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3247..3438) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(4247..5329) /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 5291..5310 /label=LacI-R /note="LacI, reverse primer" promoter complement(5330..5407) /gene="lacI" /label=lacI promoter primer_bind 5613..5632 /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 aagagaatga aattaactct taaaggagga 181 gcagctgtgg atcctgattc tggactggaa cactctgcgc atgtcctgga gaaaggtggg 241 aaggtcttca gtgccaccct tggcctggtg gacatcgtta aaggaaccaa ctcctactac 301 aagctccagc ttctggagga cgacaaggaa aacaggtatt ggatattcag gtcctggggc 361 cgtgtgggta cggtgatcgg tagcaacaaa ctggaacaga tgccgtccaa ggaggatgcc 421 attgagcact tcatgaaatt atatgaagaa aaaaccggga acgcttggca ctccaaaaat 481 ttcacgaagt atcccaaaaa gttctacccc ctggagattg actatggcca ggatgaagag 541 gcagtgaaga agctgacagt aaatcctggc taactcgagc accaccacca ccaccactga 601 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa 661 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga 721 ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg 781 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 841 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 901 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 961 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga 1021 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc 1081 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa 1141 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa 1201 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 1261 acattcaaat atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat 1321 gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa agccgtttct 1381 gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt 1441 ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa 1501 ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt 1561 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac 1621 tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat 1681 cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca 1741 gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt 1801 tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga 1861 tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat 1921 cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat 1981 acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat 2041 ataaatcagc atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa 2101 tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg 2161 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 2221 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 2281 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 2341 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta 2401 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 2461 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 2521 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 2581 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 2641 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 2701 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 2761 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 2821 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 2881 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 2941 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa 3001 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt 3061 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc 3121 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg 3181 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg 3241 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc 3301 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt 3361 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt 3421 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat 3481 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg 3541 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa 3601 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag 3661 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt 3721 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga 3781 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc 3841 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac 3901 ccgtggggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg 3961 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc 4021 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg 4081 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat 4141 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga 4201 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt 4261 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 4321 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct 4381 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc 4441 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt 4501 cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa 4561 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga 4621 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt 4681 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac 4741 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca 4801 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt 4861 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt 4921 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt 4981 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg 5041 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg 5101 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg 5161 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt 5221 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat 5281 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc 5341 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga 5401 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt 5461 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg 5521 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg 5581 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca 5641 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc 5701 tcgatcccgc gaaat //