LOCUS Exported 5118 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Bacterial expression of Nsp3 Y domain (residues 1844-1945). ACCESSION . VERSION . KEYWORDS pMCSG53_Nsp3_Y_3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5118) TITLE Savchenko Lab Plasmids REFERENCE 2 (bases 1 to 5118) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5118 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 1..25 /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 40..62 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 73..90 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 115..135 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 514..531 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(584..602) /label=T7 Term /note="T7 terminator, reverse primer" terminator 598..645 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind complement(769..788) /label=F1ori-R /note="F1 origin, reverse primer" promoter 955..1058 /gene="bla" /label=AmpR promoter CDS 1059..1919 /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(1277..1296) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2090..2678 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2579..2598 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2832..2849 /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(2929..2948) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" CDS complement(3104..4186) /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 4148..4167 /label=LacI-R /note="LacI, reverse primer" promoter complement(4187..4264) /gene="lacI" /label=lacI promoter tRNA 4404..4479 /gene="ileX" /product="E. coli tRNA-Ile recognizing AUA" /label=ileX primer_bind 4628..4647 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" tRNA 4749..4825 /gene="argU" /product="E. coli tRNA-Arg recognizing AGA and AGG" /label=argU primer_bind 5100..1 /label=T7 /note="T7 promoter, forward primer" promoter 5100..5118 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" ORIGIN 1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga 61 gatatacata tgcaccatca tcatcatcat tcttctggtg tagatctggg taccgagaac 121 ctgtacttcc aatccaatgc cgacatcgaa gttaccggtg atagctgcaa caactatatg 181 ctgacctaca acaaagtgga gaacatgacc ccgagagatt taggtgcctg catcgattgt 241 tctgcacgcc atattaatgc acaagtggca aaaagccaca acattgccct gatctggaac 301 gtgaaggact tcatgagcct gagtgaacaa ctgcgtaaac agattcgttc tgcagccaag 361 aagaacaatc tgccgttcaa actgacctgt gcaacaacac gtcaggtggt taatgttgtt 421 accaccaaga ttgccctgaa aggtggttaa cattggaagt ggataacgga tccgaattcg 481 agctccgtcg acaagcttgc ggccgcactc gagcaccacc accaccacca ctgagatccg 541 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta 601 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact 661 atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg 721 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt 781 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc 841 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg 901 gtgatggttc acattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga 961 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 1021 ccctgataaa tgcttcaata atttgaaaaa ggaagagtat gagtattcaa catttccgtg 1081 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc 1141 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg 1201 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga 1261 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc 1321 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag 1381 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga 1441 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg 1501 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga 1561 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt 1621 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact 1681 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt 1741 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg 1801 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta 1861 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac 1921 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta 1981 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt 2041 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 2101 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 2161 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 2221 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 2281 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 2341 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 2401 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 2461 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg 2521 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 2581 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 2641 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 2701 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg 2761 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa 2821 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc 2881 tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc 2941 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg tagtcatgcc 3001 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc 3061 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca 3121 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 3181 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat 3241 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca 3301 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg 3361 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg 3421 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc 3481 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc 3541 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca 3601 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg 3661 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga 3721 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca 3781 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg 3841 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca 3901 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg 3961 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca 4021 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt 4081 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag 4141 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga 4201 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg 4261 tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt 4321 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcggatt gcgacacgga 4381 gttactttat aatccgctac catggcccct tagctcagtg gttagagcag gcgactcata 4441 atcgcttggt cgctggttca agtccagcag gggccaccag atatagcaaa ggctgacgag 4501 aaatcgtcag cctttttgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc 4561 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt 4621 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggcgagat 4681 caaaagccat tgactcagca agggttgacc gtataattca cgcgattaca ccgcattgcg 4741 gtatcaacgc gcccttagct cagttggata gagcaacgac cttctaagtc gtgggccgca 4801 ggttcgaatc ctgcagggcg cgccattaca attcaatcag ttacgccttc tttatatcct 4861 ccataattcc agagtgggac atatttggga cattatcacc aaaaatgtcg tctattttcc 4921 tcgcatgctc tgtcaaatga ttaggcgcaa ggtgagcata cctacgaacc atttctatgg 4981 actcccatcc gcccatttcc tgaagcactg ataatgggac gcctgactga atccaccggt 5041 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc ccgcgaaatt 5101 aatacgactc actatagg //