LOCUS Exported 5603 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Bacterial expression of Nsp3 Macrodomain 2 and Macrodomain 3 (residues 413-676). ACCESSION . VERSION . KEYWORDS pMCSG53_Nsp3_Unique SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5603) TITLE Savchenko Lab Plasmids REFERENCE 2 (bases 1 to 5603) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5603 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 477..496 /label=LacI-R /note="LacI, reverse primer" promoter complement(516..593) /gene="lacI" /label=lacI promoter tRNA 733..808 /gene="ileX" /product="E. coli tRNA-Ile recognizing AUA" /label=ileX primer_bind 957..976 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" tRNA 1078..1154 /gene="argU" /product="E. coli tRNA-Arg recognizing AGA and AGG" /label=argU primer_bind 1429..1448 /label=T7 /note="T7 promoter, forward primer" promoter 1429..1447 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 1448..1472 /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 1487..1509 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 1520..1537 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 1562..1582 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 2446..2463 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(2516..2534) /label=T7 Term /note="T7 terminator, reverse primer" terminator 2530..2577 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind complement(2701..2720) /label=F1ori-R /note="F1 origin, reverse primer" promoter 2887..2990 /gene="bla" /label=AmpR promoter CDS 2991..3851 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(3209..3228) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4022..4610 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4511..4530 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4764..4781 /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(4861..4880) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" CDS complement(5036..515) /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 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg 61 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca 121 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg 181 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg 241 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg 301 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc 361 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc 421 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca 481 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct 541 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc 601 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc 661 gttgagcacc gccgccgcaa ggaatggtgc atgcggattg cgacacggag ttactttata 721 atccgctacc atggcccctt agctcagtgg ttagagcagg cgactcataa tcgcttggtc 781 gctggttcaa gtccagcagg ggccaccaga tatagcaaag gctgacgaga aatcgtcagc 841 ctttttgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata 901 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg 961 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggcgagatc aaaagccatt 1021 gactcagcaa gggttgaccg tataattcac gcgattacac cgcattgcgg tatcaacgcg 1081 cccttagctc agttggatag agcaacgacc ttctaagtcg tgggccgcag gttcgaatcc 1141 tgcagggcgc gccattacaa ttcaatcagt tacgccttct ttatatcctc cataattcca 1201 gagtgggaca tatttgggac attatcacca aaaatgtcgt ctattttcct cgcatgctct 1261 gtcaaatgat taggcgcaag gtgagcatac ctacgaacca tttctatgga ctcccatccg 1321 cccatttcct gaagcactga taatgggacg cctgactgaa tccaccggtg atgccggcca 1381 cgatgcgtcc ggcgtagagg atcgagatcg atctcgatcc cgcgaaatta atacgactca 1441 ctatagggga attgtgagcg gataacaatt cccctctaga aataattttg tttaacttta 1501 agaaggagat atacatatgc accatcatca tcatcattct tctggtgtag atctgggtac 1561 cgagaacctg tacttccaat ccaatatgaa aataaaggct tgtgttgaag aggtaacaac 1621 cacccttgag gaaaccaagt tcttgaccga aaacctgttg ctgtatatcg acattaacgg 1681 caacctgcac ccggacagcg cgacgttggt gagcgatatc gacattacct ttctcaaaaa 1741 agacgctccg tatatcgttg gcgatgttgt gcaggagggc gttctgaccg cggttgtcat 1801 cccgaccaaa aaggctggtg gtacgactga gatgctagcg aaggcgctgc gcaaggtgcc 1861 tacagataat tacatcacca cgtatccggg tcagggtctg aatggctaca ccgtcgagga 1921 ggccaaaacc gtgttgaaaa agtgcaaatc cgcgttttac atcctgccga gcatcatcag 1981 caatgaaaaa caggagatcc tgggtacggt gtcttggaat ctgcgtgaaa tgctggcgca 2041 cgccgaagag actcgcaagc tgatgccggt ttgtgtagaa acgaaagcca ttgtcagcac 2101 cattcaacgt aagtataaag gtattaaaat tcaagagggt gtggttgatt acggcgcaag 2161 attctacttc tataccagca aaaccaccgt cgctagcttg attaacacct taaacgatct 2221 gaacgaaact ctcgtgacca tgccactggg ctacgttacc catggtctga acttggagga 2281 ggctgcgcgt tatatgcgta gcctgaaagt gccggcaacc gtgagcgttt catctccgga 2341 cgcagttact gcgtataacg gctacctgac gtcctcgtcc taaattggaa gtggataacg 2401 gatccgaatt cgagctccgt cgacaagctt gcggccgcac tcgagcacca ccaccaccac 2461 cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct 2521 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg 2581 aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg 2641 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg 2701 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc 2761 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa 2821 aacttgatta gggtgatggt tcacattaac gcttacaatt taggtggcac ttttcgggga 2881 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc 2941 atgagacaat aaccctgata aatgcttcaa taatttgaaa aaggaagagt atgagtattc 3001 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc 3061 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt 3121 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt 3181 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg 3241 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact 3301 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg 3361 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga 3421 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg 3481 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa 3541 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac 3601 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc 3661 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca 3721 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga 3781 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta 3841 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc 3901 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc 3961 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt 4021 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac 4081 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct 4141 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact 4201 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg 4261 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata 4321 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga 4381 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag 4441 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg 4501 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac 4561 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca 4621 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg 4681 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc 4741 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga 4801 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag 4861 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac 4921 tgtagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat 4981 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact 5041 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 5101 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg 5161 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc 5221 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg 5281 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg 5341 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag 5401 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc 5461 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc 5521 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct 5581 ggtgacccaa tgcgaccaga tgc //