LOCUS Exported 4614 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp9. ACCESSION . VERSION . KEYWORDS pGBW-m4134065 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4614) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4614) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4614 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 45..116 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 121..167 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 185..233 /note="synthetic polyadenylation signal" enhancer 314..617 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 619..821 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 772..792 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 1232..1252 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1256..1321 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" polyA_signal 1374..1495 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1385..1404) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1439..1458 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 1610..1656 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 1686..1822 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 1867..1938 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(2164..2752) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2244..2263) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 3498..4358 /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(3716..3735) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 4371..4507 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" ORIGIN 1 agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag 61 ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt 121 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt 181 tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac 241 tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat 301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 601 tcatcgctat taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 781 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga 841 tcagatcttt gtcgatccta ccatccactc gacacacccg ccagcggccg ccaccatgaa 901 tgagcttagt cctgttgcac tacgacagat gtcttgtgct gccggtacta cacaaactgc 961 ttgcactgat gacaatgcgt tagcttacta caacacaaca aagggaggta ggtttgtact 1021 tgcactgtta tccgatttac aggatttgaa atgggctaga ttccctaaga gtgatggaac 1081 tggtactatc tatacagaac tggaaccacc ttgtaggttt gttacagaca cacctaaagg 1141 tcctaaagtg aagtatttat actttattaa aggattaaac aacctaaata gaggtatggt 1201 acttggtagt ttagctgcca cagtacgtct agaaaacctt tactttcagt cgatggatta 1261 taaggaccac gatggagact acaaagatca tgacattgat tacaaggacg acgatgacaa 1321 ataagctagc ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata 1381 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga 1441 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa 1501 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag 1561 caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc 1621 cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc 1681 taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca 1741 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc 1801 cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga 1861 tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca 1921 aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag 1981 tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt 2041 cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta 2101 aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 2161 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 2221 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 2281 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 2341 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 2401 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 2461 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 2521 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 2581 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 2641 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 2701 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 2761 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 2821 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 2881 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 2941 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc 3001 gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg 3061 ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt 3121 ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga 3181 gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc 3241 ggggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg 3301 ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat 3361 attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt 3421 ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat 3481 cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg 3541 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg 3601 aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc 3661 ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat 3721 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact 3781 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca 3841 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact 3901 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg 3961 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg 4021 agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg 4081 aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg 4141 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag 4201 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc 4261 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga 4321 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc 4381 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc 4441 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta 4501 tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg 4561 agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtc //