LOCUS Exported 6993 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp11. ACCESSION . VERSION . KEYWORDS pGBW-m4252914 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6993) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6993) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6993 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 47..67 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 416..433 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 526..647 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(537..556) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 591..610 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 762..808 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 838..974 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 1019..1090 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1316..1904) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1396..1415) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 2404..3613 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 3450..3469 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 3617..4645 /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 4672..4720 /note="synthetic polyadenylation signal" CDS 4986..5846 /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(5204..5223) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 5859..5995 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 6147..6218 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 6223..6269 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 6287..6335 /note="synthetic polyadenylation signal" enhancer 6416..6719 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 6721..6923 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 6874..6894 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgagt gccgatgctc agtcctttct caacggtttc gcagtggaaa acctttactt 61 tcagtcgatg agtgacaaaa tcattcatct aaccgatgac tcctttgata cagacgtgtt 121 gaaggccgat ggtgcgatac tggtagattt ctgggctgaa tggtgtggcc cgtgcaagat 181 gatcgcaccc attctggacg agatcgccga cgagtatcaa ggaaaattaa ctgtcgctaa 241 gcttaatata gatcagaacc cagggacggc acctaaatac ggcatcaggg gaatacctac 301 cctcctgctc tttaagaatg gggaagttgc cgcgacaaaa gtgggagccc tgtctaaggg 361 ccagctaaaa gagttcttgg acgctaacct ggcaggttca ggctcggggc acatgcacca 421 tcaccatcac catagcagcg ggctcgtgcc acgcggctaa ggatccgcag gcctctgcta 481 gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg 541 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg 601 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt 661 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa 721 acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc 781 ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca 841 ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag 901 attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag 961 aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca 1021 ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct 1081 tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca 1141 gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag 1201 ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt 1261 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 1321 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 1381 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 1441 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 1501 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 1561 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 1621 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 1681 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 1741 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 1801 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 1861 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 1921 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 1981 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 2041 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 2101 ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc 2161 caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct 2221 cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc 2281 ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt 2341 caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg 2401 cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg 2461 ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct tccgcccgga cgctcaggac 2521 agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt 2581 aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga 2641 ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg 2701 atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg 2761 gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg 2821 ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc 2881 gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag 2941 cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt 3001 gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgagccc 3061 ttcgctaatg cgggaaagct cttattcggg tgagatgggc tgggcaccat ctggggaccc 3121 tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag 3181 ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt 3241 cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccaccagc cggtaggtgt 3301 gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa 3361 tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc 3421 ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg 3481 gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc 3541 aatatgtaat tttcagtgtt agacttgtaa attgtccgct aaattctggc cgtttttggc 3601 ttttttgtta gacaacatgg gtaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt 3661 tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc 3721 tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc 3781 cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatccgccg cgctcccgat 3841 tccggaagtg cttgacattg gggagttcag cgagagcctg acctattgca tctcccgccg 3901 tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc 3961 ggtcgcggag gcaatggatg ccatcgctgc cgccgatctt agccagacga gcgggttcgg 4021 cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat 4081 tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt 4141 cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct 4201 cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt 4261 cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt 4321 ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc 4381 ggagcttgca ggatcgcccc ggctccgggc gtatatgctc cgcattggtc ttgaccaact 4441 ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga 4501 cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc 4561 cgccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag 4621 cactcgtccg agggcaaagg aataagctag tatgtaagcc tagtcttaga taataaaatc 4681 gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg 4741 gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt 4801 cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag 4861 cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa 4921 cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct 4981 ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc 5041 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg 5101 gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc 5161 gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat 5221 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg 5281 acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag 5341 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa 5401 cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc 5461 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca 5521 cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc 5581 tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc 5641 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg 5701 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta 5761 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag 5821 gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc 5881 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc 5941 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc 6001 cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa 6061 cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc 6121 tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt 6181 ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac 6241 ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca 6301 acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc 6361 agatctagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta 6421 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt 6481 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg 6541 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta 6601 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga 6661 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc 6721 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc 6781 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact 6841 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt 6901 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc 6961 taccatccac tcgacacacc cgccagcggc cgc //