LOCUS Exported 8055 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 surface glycoprotein (Spike protein). ACCESSION . VERSION . KEYWORDS pGBW-m4133326 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8055) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 8055) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8055 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 3824..3844 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 3845..3871 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 3924..4045 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3935..3954) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3989..4008 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 4160..4206 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 4236..4372 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 4417..4488 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(4714..5302) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4794..4813) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 6048..6908 /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(6266..6285) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 6921..7057 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 7209..7280 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 7285..7331 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 7349..7397 /note="synthetic polyadenylation signal" enhancer 7478..7781 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 7783..7985 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 7936..7956 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgttc gtctttcttg tgttactccc cctggtttcg tcccagtgcg tgaacttgac 61 tacgcgaacc caactgccgc cagcatatac aaatagtttc acccgtggag tatactaccc 121 tgataaagtg tttagatcaa gcgtactaca ctctactcag gacctgtttc tccccttctt 181 ctccaatgtc acatggtttc atgcgatcca cgttagcggc accaacggga caaagcgctt 241 tgacaaccct gtgctgccat tcaatgatgg tgtgtatttt gcttctaccg agaagagcaa 301 cataattcgg ggctggatct tcggaactac cctcgacagt aaaactcagt cattgctgat 361 tgtgaataac gccacaaatg tcgttatcaa ggtctgtgaa tttcaattct gcaacgatcc 421 attccttggg gtgtattacc ataaaaataa taagtcctgg atggagagcg aatttagggt 481 atactcgtct gccaacaatt gtactttcga gtatgtgtct cagcccttct taatggatct 541 ggaaggtaaa cagggcaact ttaagaacct ccgcgagttt gtattcaaaa atatagacgg 601 atactttaag atttattcca agcacacacc aatcaatttg gtgcgggatc tcccgcaagg 661 gttcagcgct ctggaacctc tagtggacct ccccatcggc attaacatca caagattcca 721 gacgctgtta gccctgcata gatcatacct tactcccggt gacagtagtt ctgggtggac 781 cgcaggcgcg gctgcatatt acgtgggata tttgcagcct cggacgtttc tgctgaaata 841 caacgagaac ggcaccatta cagatgccgt ggattgcgcc cttgaccccc taagcgaaac 901 taagtgtacg cttaaaagct tcaccgtcga gaagggaatt taccagactt ccaactttag 961 ggtccagccg acagaatcaa tagttaggtt cccaaatatc accaatttgt gcccttttgg 1021 ggaagtcttc aatgctacac gatttgcttc ggtttatgcc tggaacagga agcgtatctc 1081 aaattgcgtc gcggactaca gcgtgctcta taatagtgca agtttctcga cctttaaatg 1141 ttatggcgtg tccccgacca aactaaatga cctctgtttt acaaacgttt acgccgattc 1201 ttttgtgatt cgcggtgatg aggtacgcca aatcgcaccg ggccagactg gcaagattgc 1261 cgattacaac tacaagttgc ctgacgactt caccggatgt gttatagcct ggaatagcaa 1321 taatctggat tctaaggtcg gaggtaacta taactacctg tatagactgt tccgtaagtc 1381 aaacctgaaa ccctttgagc gggacatctc cacagagatt taccaagcag ggtctacccc 1441 ctgcaatggg gttgaaggat tcaattgcta ctttccatta cagagctacg ggttccagcc 1501 tacgaacggg gttggttacc agccttatag ggtagtggtg ctgtcgttcg aactgttgca 1561 cgctccagct acagtgtgcg ggcctaaaaa atcaactaac ctggttaaaa ataagtgtgt 1621 taattttaac ttcaatggct taaccggaac tggagtgctc acggagtcaa acaagaaatt 1681 tctaccgttt cagcaattcg ggagggatat cgcagacact acggacgcgg tcagagatcc 1741 acagaccctt gagatccttg acataacacc ctgctctttc ggaggtgtct ccgtaattac 1801 accaggcact aatacctcta atcaggtggc cgtgctttat caggatgtta actgtacaga 1861 ggtcccggtg gctatccatg ccgaccaatt gacacctacg tggcgcgtct attctaccgg 1921 ctccaacgtc ttccagaccc gggctggatg cttgataggc gcagaacacg taaacaactc 1981 ttacgaatgt gatattccca ttggagccgg gatctgcgca agctaccaga cccaaacaaa 2041 cagtcccaga cgggctcgta gcgtggcctc tcagagcata atagcctata ccatgtccct 2101 tggcgcggag aatagtgtgg catactcaaa taatagcatc gctatcccca ctaactttac 2161 tatttccgtg acgactgaga ttctgccagt gtctatgact aagacatccg tggattgcac 2221 aatgtatatt tgtggggatt ccacagagtg tagcaacttg ttgctgcaat atggatcatt 2281 ctgtacgcag ctgaatcgcg ccctcaccgg catcgctgtg gaacaggaca agaacactca 2341 agaggtcttt gctcaggtga agcaaatcta caagacccca cctataaaag acttcggggg 2401 ttttaacttc tcgcagattc tccccgaccc cagcaagcct tcaaaaagga gctttattga 2461 ggacctgttg ttcaacaagg ttactttagc cgatgccggt tttatcaaac agtacggcga 2521 ctgtcttggc gatatagcag cccgggatct gatttgcgca caaaaattca atggccttac 2581 cgtactgcca ccccttctca ccgacgaaat gatcgcgcag tacacctccg cactgctggc 2641 tgggaccatc acttcgggat ggacctttgg ggctggtgct gctttgcaaa ttcccttcgc 2701 catgcagatg gcctatcgat tcaatggcat aggggtaaca cagaacgtat tgtatgaaaa 2761 tcaaaagcta atcgctaacc agtttaattc agccattggt aagatccagg attcactctc 2821 cagtacagcc tctgccctgg gaaagctcca ggacgtggtc aaccagaacg cgcaggcatt 2881 gaatacactc gtcaaacagc tgagtagcaa tttcggcgca atcagtagtg tcttaaacga 2941 tatactgagt aggcttgata aagtagaggc cgaggtccag attgacagac taatcacggg 3001 gcgtcttcag agtttacaga cctatgtgac ccaacagctg atccgagcag cggaaattcg 3061 agcaagcgcg aacctggccg ctactaaaat gagtgagtgc gtgctgggcc aaagtaaacg 3121 agtcgacttt tgcggcaaag ggtatcatct catgtctttc cctcagtccg cccctcacgg 3181 tgttgttttc ctccacgtga catacgttcc cgcgcaggaa aagaatttta ctacggcacc 3241 cgctatttgc catgacggaa aggctcattt tccacgggag ggagtgttcg tttcaaacgg 3301 gacacactgg tttgtaactc agcggaactt ttatgagcct caaatcatca cgacagacaa 3361 taccttcgtg agtggcaact gcgatgtagt cataggcatt gtgaataaca ctgtttatga 3421 tccacttcag ccggaactcg acagcttcaa agaggaattg gataagtact ttaaaaatca 3481 taccagtcca gacgtagatc tcggcgatat atcgggcatt aacgccagtg tggtcaatat 3541 acaaaaggag attgacagac tgaatgaagt agctaagaac ctcaacgaat ctctgattga 3601 cctgcaagag ctagggaagt acgagcagta tatcaaatgg ccatggtaca tatggctggg 3661 ttttatcgcg ggtcttattg ccatagtcat ggtgacgatt atgttgtgct gtatgacctc 3721 ctgctgttct tgtttgaaag gctgctgctc ctgtggtagc tgttgcaaat tcgatgaaga 3781 tgactctgaa cccgtgttga aaggcgttaa gctacactac acagaaaacc tttactttca 3841 gtcgtacccc tatgacgtac cagattatgc gtaagctagc ttgactgact gagatacagc 3901 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag 3961 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac 4021 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt 4081 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg 4141 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt 4201 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg 4261 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc 4321 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca 4381 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg 4441 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt 4501 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg 4561 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg 4621 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc 4681 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 4741 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 4801 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 4861 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 4921 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 4981 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 5041 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 5101 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 5161 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 5221 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 5281 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 5341 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 5401 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 5461 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 5521 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct 5581 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc 5641 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa 5701 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga 5761 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggtgtaccg agaacgatcc 5821 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag 5881 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta 5941 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag 6001 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac 6061 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc 6121 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca 6181 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc 6241 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg 6301 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac 6361 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca 6421 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg 6481 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac 6541 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg 6601 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt 6661 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg 6721 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg 6781 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc 6841 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc 6901 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg 6961 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact 7021 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga 7081 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt 7141 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg 7201 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac 7261 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag 7321 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga 7381 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta 7441 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 7501 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 7561 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 7621 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 7681 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 7741 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt 7801 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 7861 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 7921 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 7981 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca 8041 cccgccagcg gccgc //