LOCUS Exported 7452 bp ds-DNA circular SYN 13-APR-2022 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp6. ACCESSION . VERSION . KEYWORDS pGBW-m4133604 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7452) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 7452) AUTHORS . TITLE Direct Submission JOURNAL Exported Apr 13, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7452 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 878..898 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 899..916 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 985..1106 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(996..1015) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1050..1069 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 1221..1267 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 1297..1433 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 1478..1549 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1775..2363) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1855..1874) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 2863..4072 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 3909..3928 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 4076..5104 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA KE" polyA_signal 5131..5179 /note="synthetic polyadenylation signal" CDS 5445..6305 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(5663..5682) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 6318..6454 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 6606..6677 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 6682..6728 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 6746..6794 /note="synthetic polyadenylation signal" enhancer 6875..7178 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 7180..7382 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 7333..7353 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgagc gccgttaagc ggacaatcaa agggacccat cactggttgc tccttactat 61 actgacgtct ttactggtct tggtacagag tacacaatgg tcgcttttct tttttctgta 121 cgagaatgca ttcctcccat ttgcgatggg aatcattgct atgtccgcct tcgctatgat 181 gttcgtgaag cacaaacatg ccttcctttg cctgtttctc ctaccctcac tggcaaccgt 241 ggcctatttt aacatggtgt acatgcctgc gagctgggtc atgcgaatta tgacttggct 301 ggacatggtt gatacctccc tatctggctt caagttaaag gactgtgtga tgtatgcatc 361 agctgtagtg ctgctaatcc tgatgacagc tcgcacagtc tacgatgacg gtgccaggag 421 agtgtggact ctgatgaatg tcctcacctt agtttataaa gtatactatg gcaacgctct 481 cgatcaggcc attagtatgt gggcattgat aatctccgtg acgagtaact acagcggagt 541 ggtaaccact gtcatgtttt tggcacgcgg gatagtgttc atgtgtgtcg aatattgccc 601 gatctttttt atcacaggga atactctgca atgcattatg ctggtttact gtttcttggg 661 ctatttctgc acctgttatt ttggtctatt ctgcctgctt aataggtact ttcgtttgac 721 actaggagtc tatgattact tggtgagcac gcaggagttt cggtatatga acagtcaggg 781 cctgcttccc ccaaagaact ctattgacgc gtttaaactc aatataaagc tgttgggcgt 841 gggtggaaaa ccttgcatca aggtggccac tgtgcaggaa aacctttact ttcagtcgca 901 ccatcaccat caccattaag gatccgcagg cctctgctag cttgactgac tgagatacag 961 cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa accacaacta 1021 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa 1081 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg 1141 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg 1201 gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg 1261 ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct 1321 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt 1381 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc 1441 agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg 1501 gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt 1561 taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt 1621 gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt 1681 gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg 1741 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 1801 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 1861 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 1921 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 1981 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 2041 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 2101 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 2161 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 2221 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 2281 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 2341 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 2401 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 2461 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 2521 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 2581 ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc 2641 ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct 2701 cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca 2761 atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg 2821 atctatccga gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt 2881 tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc 2941 gcaggagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga 3001 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc 3061 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg 3121 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc 3181 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga 3241 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc 3301 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt 3361 ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt 3421 cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg 3481 ggggcatggt gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc 3541 ttattcgggt gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga 3601 ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca 3661 gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt 3721 ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg 3781 caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg 3841 gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct 3901 taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga 3961 agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta 4021 gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatggg 4081 taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag 4141 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt 4201 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg 4261 ttatgtttat cggcactttg catccgccgc gctcccgatt ccggaagtgc ttgacattgg 4321 ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca 4381 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg caatggatgc 4441 catcgctgcc gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat 4501 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca 4561 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct 4621 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc 4681 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat 4741 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg 4801 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccccg 4861 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg 4921 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc 4981 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga ccgatggctg 5041 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga 5101 ataagctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg 5161 tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct 5221 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt 5281 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg 5341 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt 5401 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt 5461 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag 5521 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg 5581 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa 5641 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc 5701 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag 5761 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa 5821 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc 5881 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg 5941 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa 6001 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga 6061 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg 6121 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag 6181 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg 6241 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt 6301 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct 6361 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc 6421 atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta aactcgactc 6481 tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct 6541 cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct 6601 cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc 6661 agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg 6721 ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg 6781 atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg 6841 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc 6901 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca 6961 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg 7021 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg 7081 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt 7141 ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt tggcagtaca 7201 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg 7261 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact 7321 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag 7381 ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact cgacacaccc 7441 gccagcggcc gc //