LOCUS Exported 9962 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 S (surface glycoprotein). ACCESSION . VERSION . KEYWORDS pGBW-m4134154 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9962) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 9962) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9962 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 138..209 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 214..260 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 278..326 /note="synthetic polyadenylation signal" enhancer 407..710 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 712..914 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 865..885 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 4808..4828 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 4829..4855 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 4908..5029 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4919..4938) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 4973..4992 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 5144..5190 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 5220..5356 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 5401..5472 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(5698..6286) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5778..5797) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 6786..7995 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 7832..7851 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 7999..8598 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(7999..8018) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 8495..8515 /label=Puro-F /note="Puromycin resistance gene, forward primer" polyA_signal 8625..8673 /note="synthetic polyadenylation signal" CDS 8939..9799 /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(9157..9176) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 9812..9948 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" ORIGIN 1 taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg 61 cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca 121 gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt 181 ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga 241 ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc 301 catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt 361 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta 421 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga 481 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt 541 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta 601 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg 661 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt 721 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc 781 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat 841 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct 901 atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca 961 ctcgacacac ccgccagcgg ccgccaccat gttcgtcttt cttgtgttac tccccctggt 1021 ttcgtcccag tgcgtgaact tgactacgcg aacccaactg ccgccagcat atacaaatag 1081 tttcacccgt ggagtatact accctgataa agtgtttaga tcaagcgtac tacactctac 1141 tcaggacctg tttctcccct tcttctccaa tgtcacatgg tttcatgcga tccacgttag 1201 cggcaccaac gggacaaagc gctttgacaa ccctgtgctg ccattcaatg atggtgtgta 1261 ttttgcttct accgagaaga gcaacataat tcggggctgg atcttcggaa ctaccctcga 1321 cagtaaaact cagtcattgc tgattgtgaa taacgccaca aatgtcgtta tcaaggtctg 1381 tgaatttcaa ttctgcaacg atccattcct tggggtgtat taccataaaa ataataagtc 1441 ctggatggag agcgaattta gggtatactc gtctgccaac aattgtactt tcgagtatgt 1501 gtctcagccc ttcttaatgg atctggaagg taaacagggc aactttaaga acctccgcga 1561 gtttgtattc aaaaatatag acggatactt taagatttat tccaagcaca caccaatcaa 1621 tttggtgcgg gatctcccgc aagggttcag cgctctggaa cctctagtgg acctccccat 1681 cggcattaac atcacaagat tccagacgct gttagccctg catagatcat accttactcc 1741 cggtgacagt agttctgggt ggaccgcagg cgcggctgca tattacgtgg gatatttgca 1801 gcctcggacg tttctgctga aatacaacga gaacggcacc attacagatg ccgtggattg 1861 cgcccttgac cccctaagcg aaactaagtg tacgcttaaa agcttcaccg tcgagaaggg 1921 aatttaccag acttccaact ttagggtcca gccgacagaa tcaatagtta ggttcccaaa 1981 tatcaccaat ttgtgccctt ttggggaagt cttcaatgct acacgatttg cttcggttta 2041 tgcctggaac aggaagcgta tctcaaattg cgtcgcggac tacagcgtgc tctataatag 2101 tgcaagtttc tcgaccttta aatgttatgg cgtgtccccg accaaactaa atgacctctg 2161 ttttacaaac gtttacgccg attcttttgt gattcgcggt gatgaggtac gccaaatcgc 2221 accgggccag actggcaaga ttgccgatta caactacaag ttgcctgacg acttcaccgg 2281 atgtgttata gcctggaata gcaataatct ggattctaag gtcggaggta actataacta 2341 cctgtataga ctgttccgta agtcaaacct gaaacccttt gagcgggaca tctccacaga 2401 gatttaccaa gcagggtcta ccccctgcaa tggggttgaa ggattcaatt gctactttcc 2461 attacagagc tacgggttcc agcctacgaa cggggttggt taccagcctt atagggtagt 2521 ggtgctgtcg ttcgaactgt tgcacgctcc agctacagtg tgcgggccta aaaaatcaac 2581 taacctggtt aaaaataagt gtgttaattt taacttcaat ggcttaaccg gaactggagt 2641 gctcacggag tcaaacaaga aatttctacc gtttcagcaa ttcgggaggg atatcgcaga 2701 cactacggac gcggtcagag atccacagac ccttgagatc cttgacataa caccctgctc 2761 tttcggaggt gtctccgtaa ttacaccagg cactaatacc tctaatcagg tggccgtgct 2821 ttatcaggat gttaactgta cagaggtccc ggtggctatc catgccgacc aattgacacc 2881 tacgtggcgc gtctattcta ccggctccaa cgtcttccag acccgggctg gatgcttgat 2941 aggcgcagaa cacgtaaaca actcttacga atgtgatatt cccattggag ccgggatctg 3001 cgcaagctac cagacccaaa caaacagtcc cagacgggct cgtagcgtgg cctctcagag 3061 cataatagcc tataccatgt cccttggcgc ggagaatagt gtggcatact caaataatag 3121 catcgctatc cccactaact ttactatttc cgtgacgact gagattctgc cagtgtctat 3181 gactaagaca tccgtggatt gcacaatgta tatttgtggg gattccacag agtgtagcaa 3241 cttgttgctg caatatggat cattctgtac gcagctgaat cgcgccctca ccggcatcgc 3301 tgtggaacag gacaagaaca ctcaagaggt ctttgctcag gtgaagcaaa tctacaagac 3361 cccacctata aaagacttcg ggggttttaa cttctcgcag attctccccg accccagcaa 3421 gccttcaaaa aggagcttta ttgaggacct gttgttcaac aaggttactt tagccgatgc 3481 cggttttatc aaacagtacg gcgactgtct tggcgatata gcagcccggg atctgatttg 3541 cgcacaaaaa ttcaatggcc ttaccgtact gccacccctt ctcaccgacg aaatgatcgc 3601 gcagtacacc tccgcactgc tggctgggac catcacttcg ggatggacct ttggggctgg 3661 tgctgctttg caaattccct tcgccatgca gatggcctat cgattcaatg gcataggggt 3721 aacacagaac gtattgtatg aaaatcaaaa gctaatcgct aaccagttta attcagccat 3781 tggtaagatc caggattcac tctccagtac agcctctgcc ctgggaaagc tccaggacgt 3841 ggtcaaccag aacgcgcagg cattgaatac actcgtcaaa cagctgagta gcaatttcgg 3901 cgcaatcagt agtgtcttaa acgatatact gagtaggctt gataaagtag aggccgaggt 3961 ccagattgac agactaatca cggggcgtct tcagagttta cagacctatg tgacccaaca 4021 gctgatccga gcagcggaaa ttcgagcaag cgcgaacctg gccgctacta aaatgagtga 4081 gtgcgtgctg ggccaaagta aacgagtcga cttttgcggc aaagggtatc atctcatgtc 4141 tttccctcag tccgcccctc acggtgttgt tttcctccac gtgacatacg ttcccgcgca 4201 ggaaaagaat tttactacgg cacccgctat ttgccatgac ggaaaggctc attttccacg 4261 ggagggagtg ttcgtttcaa acgggacaca ctggtttgta actcagcgga acttttatga 4321 gcctcaaatc atcacgacag acaatacctt cgtgagtggc aactgcgatg tagtcatagg 4381 cattgtgaat aacactgttt atgatccact tcagccggaa ctcgacagct tcaaagagga 4441 attggataag tactttaaaa atcataccag tccagacgta gatctcggcg atatatcggg 4501 cattaacgcc agtgtggtca atatacaaaa ggagattgac agactgaatg aagtagctaa 4561 gaacctcaac gaatctctga ttgacctgca agagctaggg aagtacgagc agtatatcaa 4621 atggccatgg tacatatggc tgggttttat cgcgggtctt attgccatag tcatggtgac 4681 gattatgttg tgctgtatga cctcctgctg ttcttgtttg aaaggctgct gctcctgtgg 4741 tagctgttgc aaattcgatg aagatgactc tgaacccgtg ttgaaaggcg ttaagctaca 4801 ctacacagaa aacctttact ttcagtcgta cccctatgac gtaccagatt atgcgtaagc 4861 tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga 4921 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg 4981 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa 5041 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta 5101 aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg 5161 gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg 5221 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc 5281 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc 5341 agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga 5401 cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt 5461 cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt 5521 cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca 5581 agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat 5641 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 5701 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 5761 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 5821 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 5881 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 5941 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 6001 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 6061 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 6121 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt 6181 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 6241 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 6301 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 6361 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 6421 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc 6481 acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc 6541 tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct 6601 ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc 6661 gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag 6721 ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg 6781 cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag 6841 cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg 6901 acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt 6961 ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc 7021 gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc 7081 cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt 7141 gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg 7201 ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg tgtggagaga 7261 ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg ggttgggggg 7321 agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt gaggcgggct 7381 gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa ggtcttgagc 7441 ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc atctggggac 7501 cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg cgggggcggc 7561 agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt 7621 gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacct gccggtaggt 7681 gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta ggctctcctg 7741 aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca gtttctttgg 7801 tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg 7861 gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta atcatttggg 7921 tcaatatgta attttcagtg ttagacttgt aaattgtccg ctaaattctg gccgtttttg 7981 gcttttttgt tagacaacat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac 8041 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc 8101 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc 8161 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg 8221 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc 8281 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaggg cctcctggcg 8341 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac 8401 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggccgc cgagcgcgcc 8461 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc 8521 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc 8581 cgcaagcccg gtgcctaggc tagtatgtaa gcctagtctt agataataaa atcgctatcc 8641 atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc 8701 gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag 8761 tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc 8821 acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa 8881 tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat 8941 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt 9001 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg 9061 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga 9121 agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg 9181 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt 9241 tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg 9301 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg 9361 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga 9421 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc 9481 tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc 9541 ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc 9601 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg 9661 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac 9721 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc 9781 actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc 9841 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca 9901 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga tcccgaatcg 9961 tt //