LOCUS Exported 4935 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp9. ACCESSION . VERSION . KEYWORDS pGBW-m4133355 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4935) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4935) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4935 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 341..361 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 710..727 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 804..925 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(815..834) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 869..888 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 1040..1086 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 1116..1252 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 1297..1368 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1594..2182) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1674..1693) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2928..3788 /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(3146..3165) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 3801..3937 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 4089..4160 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 4165..4211 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 4229..4277 /note="synthetic polyadenylation signal" enhancer 4358..4661 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4663..4865 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4816..4836 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgaat gagcttagtc ctgttgcact acgacagatg tcttgtgctg ccggtactac 61 acaaactgct tgcactgatg acaatgcgtt agcttactac aacacaacaa agggaggtag 121 gtttgtactt gcactgttat ccgatttaca ggatttgaaa tgggctagat tccctaagag 181 tgatggaact ggtactatct atacagaact ggaaccacct tgtaggtttg ttacagacac 241 acctaaaggt cctaaagtga agtatttata ctttattaaa ggattaaaca acctaaatag 301 aggtatggta cttggtagtt tagctgccac agtacgtcta gaaaaccttt actttcagtc 361 gatgagtgac aaaatcattc atctaaccga tgactccttt gatacagacg tgttgaaggc 421 cgatggtgcg atactggtag atttctgggc tgaatggtgt ggcccgtgca agatgatcgc 481 acccattctg gacgagatcg ccgacgagta tcaaggaaaa ttaactgtcg ctaagcttaa 541 tatagatcag aacccaggga cggcacctaa atacggcatc aggggaatac ctaccctcct 601 gctctttaag aatggggaag ttgccgcgac aaaagtggga gccctgtcta agggccagct 661 aaaagagttc ttggacgcta acctggcagg ttcaggctcg gggcacatgc accatcacca 721 tcaccatagc agcgggctcg tgccacgcgg ctaagctagc ttgactgact gagatacagc 781 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag 841 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac 901 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt 961 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg 1021 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt 1081 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg 1141 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc 1201 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca 1261 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg 1321 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt 1381 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg 1441 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg 1501 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc 1561 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 1621 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 1681 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 1741 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 1801 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 1861 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 1921 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 1981 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 2041 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 2101 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 2161 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 2221 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 2281 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 2341 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 2401 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct 2461 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc 2521 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa 2581 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga 2641 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggtgtaccg agaacgatcc 2701 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag 2761 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta 2821 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag 2881 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac 2941 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc 3001 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca 3061 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc 3121 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg 3181 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac 3241 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca 3301 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg 3361 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac 3421 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg 3481 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt 3541 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg 3601 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg 3661 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc 3721 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc 3781 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg 3841 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact 3901 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga 3961 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt 4021 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg 4081 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac 4141 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag 4201 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga 4261 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta 4321 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 4381 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 4441 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 4501 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 4561 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 4621 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt 4681 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 4741 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 4801 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 4861 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca 4921 cccgccagcg gccgc //