LOCUS Exported 4596 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp9. ACCESSION . VERSION . KEYWORDS pGBW-m4133936 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4596) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4596) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4596 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 347..367 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 368..412 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" polyA_signal 465..586 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(476..495) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 530..549 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 701..747 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 777..913 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 958..1029 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1255..1843) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1335..1354) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2589..3449 /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(2807..2826) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 3462..3598 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 3750..3821 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 3826..3872 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 3890..3938 /note="synthetic polyadenylation signal" enhancer 4019..4322 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4324..4526 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4477..4497 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgaat aacgagttaa gcccagtagc gcttcgccag atgagttgcg ccgcaggtac 61 aacccaaact gcttgtacgg atgacaacgc cctcgcttac tataatacca caaagggagg 121 ccgattcgtt ctggccctgt tgtcggatct gcaagacttg aaatgggcac gttttcccaa 181 gtccgacggg actggcacca tttacacaga actagagccg ccatgcaggt tcgtgactga 241 tacccctaaa ggacccaagg tcaaatatct gtattttatc aaggggctta acaatctcaa 301 tagaggcatg gtgttaggat ctctagccgc aacggtgcgg ctgcaagaaa acctttactt 361 tcagtcgaaa gaaaccgcgg cagctaagtt cgagaggcag cacatggaca gttaagctag 421 cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat acattgatga 481 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga 541 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg 601 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa 661 cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc 721 tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat 781 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga 841 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga 901 aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat 961 tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt 1021 ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag 1081 ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt 1141 tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg 1201 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 1261 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 1321 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 1381 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 1441 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 1501 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 1561 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 1621 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 1681 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 1741 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 1801 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 1861 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 1921 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 1981 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 2041 tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc 2101 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc 2161 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc 2221 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc 2281 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc 2341 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc 2401 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc 2461 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt 2521 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag 2581 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt 2641 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt 2701 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt 2761 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg 2821 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga 2881 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa 2941 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga 3001 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa 3061 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca 3121 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta 3181 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac 3241 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc 3301 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag 3361 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga 3421 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa 3481 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac 3541 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga 3601 tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc 3661 gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt 3721 acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt 3781 cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa 3841 gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa 3901 ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta 3961 gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 4021 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 4081 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 4141 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 4201 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 4261 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 4321 tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat 4381 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 4441 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac 4501 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga 4561 tcctaccatc cactcgacac acccgccagc ggccgc //