LOCUS Exported 7035 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 RNA-dependent RNA polymerase. ACCESSION . VERSION . KEYWORDS pGBW-m4133914 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7035) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 7035) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7035 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 2804..2824 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 2825..2851 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 2904..3025 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2915..2934) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2969..2988 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 3140..3186 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 3216..3352 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 3397..3468 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(3694..4282) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3774..3793) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 5028..5888 /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(5246..5265) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 5901..6037 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 6189..6260 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 6265..6311 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 6329..6377 /note="synthetic polyadenylation signal" enhancer 6458..6761 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 6763..6965 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 6916..6936 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgtct gccgacgcac agtcgttcct gaatagagtt tgtggtgtat ccgcggcccg 61 gctaactcca tgcggaaccg ggacgagcac agatgtggtg taccgagctt ttgatatcta 121 taacgacaaa gtcgctggct tcgccaagtt tcttaagacc aactgttgca ggttccaaga 181 gaaagacgaa gatgataatc tcatagactc atactttgtg gttaagcgtc acacttttag 241 taattatcag catgaggaaa caatttataa cttgctgaag gactgccctg ccgtagcaaa 301 acacgacttc tttaaattcc gcatcgatgg cgatatggtc ccgcatatta gcagacagcg 361 gttaaccaag tacactatgg cagacctggt gtacgccctg cgacacttcg acgagggaaa 421 ctgtgacacg ttaaaagaaa tcctcgtgac ttataattgt tgcgatgacg attatttcaa 481 caagaaagat tggtacgatt ttgtcgagaa tcccgatata ctcagagtgt acgctaacct 541 gggggaaagg gttcgccagg cactattgaa gacagtccaa ttttgcgacg cgatgcggaa 601 tgccggcatt gtgggtgtcc tgacacttga taaccaggac ctgaatggca actggtatga 661 ctttggagat ttcattcaga ccacgcccgg gtccggagta cctgtcgttg actcttacta 721 ctcacttctg atgccaatcc taaccttaac ccgagctctc acagctgaga gtcacgtgga 781 caccgacctc acaaaaccgt atatcaagtg ggatttgctg aagtacgatt tcacggaaga 841 gaggttaaag ctctttgatc gttatttcaa atactgggac cagacttatc atcccaactg 901 tgtgaattgt ctggacgaca gatgcatact tcattgtgcc aatttcaatg tactgtttag 961 cacagtgttc ccaccaactt cgtttggtcc tctagtgcgc aaaatctttg tggatggggt 1021 gcccttcgtt gtttccacag gctatcactt tcgtgagctc ggcgtagttc acaaccaaga 1081 tgtcaacctg cattcttcca ggttaagttt taaggaactt ctggtgtatg ccgcagaccc 1141 tgcgatgcac gctgcaagcg gtaatttgtt gctggataag cggaccacgt gcttcagcgt 1201 ggcggcttta acaaataacg ttgccttcca aaccgtcaag cctggcaact tcaataaaga 1261 tttctacgat ttcgccgtca gtaaagggtt ttttaaggag ggatcaagcg tagaattgaa 1321 acattttttc tttgctcagg acggtaatgc cgctatctcg gactacgact actaccggta 1381 taacctacca actatgtgcg acattcggca gctgttgttc gtggtggagg tcgttgataa 1441 atactttgat tgctacgatg gcggatgcat taacgccaat caagtcatcg tgaacaatct 1501 ggacaagagc gccggcttcc cctttaacaa atggggaaag gcaaggctat attatgactc 1561 tatgtcctat gaggatcagg acgctctctt cgcgtatacc aagcgcaacg ttatccccac 1621 aattacccag atgaacctca aatatgcaat cagcgcaaag aacagagcga ggacagttgc 1681 tggggtgagt atatgttcga caatgactaa caggcagttt caccagaagc tgttgaaatc 1741 tattgccgct acacgagggg ctaccgtcgt gattggcacc tccaaattct acggagggtg 1801 gcataatatg ttaaaaactg tgtactccga cgtggaaaac cctcacctga tgggttggga 1861 ttatcctaag tgcgaccggg cgatgccaaa catgctgaga attatggcgt cattagtcct 1921 ggcgcgaaag cacaccacgt gctgtagtct gtcacatcgc ttttacagat tggctaatga 1981 atgcgcccaa gtgcttagcg agatggtgat gtgtgggggt tcactttacg tgaagcccgg 2041 aggaacgtcc tcaggcgacg ccacaactgc ctacgcaaat tcagtattca acatatgtca 2101 ggccgttaca gcaaacgtga atgctctgct gtctactgat gggaacaaga tcgccgataa 2161 gtacgtccgg aacctgcaac acaggctgta cgagtgtctg tatcgtaatc gagatgtaga 2221 caccgacttc gtgaacgaat tttacgcata cttacggaaa catttctcca tgatgatcct 2281 ctccgacgat gcagttgtgt gctttaattc aacttacgcc tctcagggct tggtggccag 2341 tatcaaaaat tttaagtcgg ttctttatta tcagaataac gtgtttatga gcgaggctaa 2401 gtgttggacc gagactgatc tgaccaaagg cccgcacgag ttctgttccc agcacaccat 2461 gctggtcaag cagggtgatg actatgtcta tttaccatac ccagatccga gtaggatctt 2521 gggagccggt tgttttgtgg atgacattgt gaagactgac ggtacactca tgattgagcg 2581 cttcgtcagt ttggcaatcg acgcttaccc ccttaccaaa catccaaatc aggaatatgc 2641 ggatgtgttt cacctgtacc ttcaatacat aaggaaactt catgatgaac taactggaca 2701 tatgctcgac atgtattctg taatgcttac aaatgacaat acctcacggt actgggaacc 2761 ggagttttat gaagccatgt acaccccaca tactgtcctg caagaaaacc tttactttca 2821 gtcgtacccc tatgacgtac cagattatgc gtaagctagc ttgactgact gagatacagc 2881 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag 2941 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac 3001 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt 3061 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg 3121 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt 3181 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg 3241 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc 3301 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca 3361 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg 3421 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt 3481 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg 3541 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg 3601 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc 3661 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 3721 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 3781 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 3841 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 3901 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 3961 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 4021 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 4081 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 4141 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 4201 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 4261 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 4321 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 4381 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 4441 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 4501 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct 4561 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc 4621 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa 4681 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga 4741 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggtgtaccg agaacgatcc 4801 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag 4861 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta 4921 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag 4981 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac 5041 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc 5101 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca 5161 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc 5221 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg 5281 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac 5341 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca 5401 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg 5461 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac 5521 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg 5581 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt 5641 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg 5701 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg 5761 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc 5821 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc 5881 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg 5941 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact 6001 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga 6061 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt 6121 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg 6181 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac 6241 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag 6301 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga 6361 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta 6421 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 6481 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 6541 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 6601 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 6661 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 6721 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt 6781 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 6841 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 6901 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 6961 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca 7021 cccgccagcg gccgc //