LOCUS Exported 6299 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 ORF10 protein. ACCESSION . VERSION . KEYWORDS pGBW-m4252818 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6299) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6299) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6299 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 31..79 /note="synthetic polyadenylation signal" terminator 97..143 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" misc_feature 148..219 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 371..507 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" CDS complement(520..1380) /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 1143..1162 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" polyA_signal 1646..1694 /note="synthetic polyadenylation signal" CDS complement(1721..2320) /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(1804..1824) /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind 2301..2320 /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" promoter complement(2324..3533) /label=UbC promoter /note="human ubiquitin C promoter" primer_bind complement(2468..2487) /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" rep_origin 4033..4621 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4522..4541 /label=pBR322ori-F /note="pBR322 origin, forward primer" misc_feature 4847..4918 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" terminator 4963..5099 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" terminator 5129..5175 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 5290..5411 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5327..5346) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5381..5400 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" CDS complement(5464..5529) /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS complement(5533..5553) /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" regulatory 5664..5673 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter complement(5742..5944) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(5771..5791) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 5946..6249 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 gatctggcta cactcgatca gcagttagat aataaaatcg ctatccatcg aagatggatg 61 tgtgttggtt ttttgtgtgt gtaacgcaac gattgatagc ataacccctt ggggcctcta 121 aacgggtctt gaggggtttt ttgaattgag aggtagctga agctgctgtt atgaccacta 181 gagggaagaa gatacctgtg gagctaatgg tccaagatgt cgggagccgc gatcgctgcc 241 aaaaaggtaa gctgacgata gctgacgaga ttcgatgccc gacgagcaaa tgagcgccac 301 ggctgttcgc gtaagctcga aacgacggag attcgataga gccagagtcg agtttaaacg 361 attcgggatc ttaaggataa atttctggta aggaggacac gtatggaagt gggcaagttg 421 gggaagccgt atccgttgct gaatctggca tatgtgggag tataagacgc gcagcgtcgc 481 atcaggcatt tttttctgcg ccaatgcacc tagaatcggt taccaatgct taatcagtga 541 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 601 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 661 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 721 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctatcaact gttgccggga 781 agctagagta agtagttcgc cagttaatag tttacgcaag gttgttgcca ttgctacagg 841 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 901 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 961 aatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 1021 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg aatactcaac 1081 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 1141 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa agcgttcttc 1201 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 1261 cgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 1321 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 1381 gcgaaagcct cctgctgatc ctgtctcttg atagatgttt gcaaaagcta ggactcgatt 1441 atacgttgac cgatcagact atcaatatct aggtttaaac agatcggtac gctgccgtga 1501 ccaggcttga gtacaatatc tgacgattgg acttcctggg tcccaagctg gattccgact 1561 ggctgactgc caagcaacga tatggatcgt cgagactcgc actgagagga tcgttctcgg 1621 tacaccaggc gcgcctagcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg 1681 atagcgattt tattatctaa gactaggctt acatactagc ctaggcaccg ggcttgcggg 1741 tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga 1801 gccgctcgta gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg 1861 cgcgctcggc ggcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt 1921 ggtcgggcga gacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg 1981 ccaggaggcc ctccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc 2041 gcgggccgat ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg 2101 ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga 2161 ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg acggtgtggc 2221 gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt 2281 cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat gttgtctaac aaaaaagcca 2341 aaaacggcca gaatttagcg gacaatttac aagtctaaca ctgaaaatta catattgacc 2401 caaatgatta catttcaaaa ggtgcctaaa aaacttcaca aaacacactc gccaaccccg 2461 agcgcatagt tcaaaaccgg agcttcagct acttaagaag ataggtacat aaaaccgacc 2521 aaagaaactg acgcctcact tatccctccc ctcaccagag gtccggcgcc tgtcgattca 2581 ggagagccta ccctaggccc gaaccctgcg tcctgcgacg gagaaaagcc taccgcacac 2641 ctaccggctg gtggccccac cctgcattat aagccaacag aacgggtgac gtcacgacac 2701 gacgagggcg cgcgctccca aaggtacggg tgcactgccc aacggcaccg ccataactgc 2761 cgcccccgca acagacgaca aaccgagttc tccagtcagt gacaaacttc acgtcagggt 2821 ccccagatgg tgcccagccc atctcacccg aataagagct ttcccgcatt agcgaagggc 2881 tcaagacctt gggttcttgc cgcccaccat gccccccacc ttgtttcaac gacctcacag 2941 cccgcctcac aagcgtcttc cattcaagac tcgggaacag ccgccatttt gctgcgctcc 3001 ccccaacccc cagttcaggg caaccttgct cgcggaccca gactacagcc cttggcggtc 3061 tctccacacg cttccgtccc accgagcggc ccggcggcca cgaaagcccc ggccagccca 3121 gcagcccgct actcaccaag tgacgatcac agcgatccac aaacaagaac cgcgacccaa 3181 atcccggctg cgacggaact agctgtgcca cacccggcgc gtccttatat aatcatcggc 3241 gttcaccgcc ccacggagat ccctccgcag aatcgccgag aagggactac ttttcctcgc 3301 ctgttccgct ctctggaaag aaaaccagtg ccctagagtc acccaagtcc cgtcctaaaa 3361 tgtccttctg ctgatactgg ggttctaagg ccgagtctta tgagcagcgg gccgctgtcc 3421 tgagcgtccg ggcggaagga tcaggacgct cctgcgccct tcgtctgacg tggcagcgct 3481 cgccgtgacg aggggggcgc ccgcgggagg cgccaaaacc cggcgcggag gccaggcgcg 3541 ccccgatttc gatattcctc ggatctcgga tagatcggga ttgaggatgt aggttgaact 3601 tctctcgggt gatcccgatc tcggatattc gggattggag taatacctgg cgataggcgc 3661 tgccaagcaa tagccaaggc gcaaggccgg ccaagagact tgcgatagtt cgttcgagag 3721 gtagcctgca agatggatcg gacgaaaggt ttgaagattt tccgacgagg gccttggaga 3781 gttcggaaat ttaaatgcaa ctatggatga acgaaataga cagatcgctg agataggtgc 3841 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga 3901 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat 3961 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat 4021 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa 4081 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa 4141 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt 4201 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt 4261 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata 4321 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt 4381 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac 4441 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga 4501 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg 4561 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa 4621 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat 4681 gatttaaatg aaactcgtaa tcggagaacg ggtgcaatag caaggaccag atcgaacttg 4741 actgacgcaa ttctgagcgt agtcgaaacc cagacaagtc gatctcgcca agtagctgac 4801 ttgactggtt aattaagttt cgatagccca aggtaaccaa tgagattagg gagggaagaa 4861 agtttgatga actacttctg acccctagtg gtgtccagaa aagaccatta aaggaatgtc 4921 ttgatcgaaa tcgagagatc gcctgcagga tagctgacga ccttaaggat aaatttctgg 4981 taaggaggac acgtatggaa gtgggcaagt tggggaagcc gtatccgttg ctgaatctgg 5041 catatgtggg agtataagac gcgcagcgtc gcatcaggca tttttttctg cgccaatgcc 5101 ggtagatagc aatccggccc gaggggcaca aaaaacccct caagacccgt ttagaggccc 5161 caaggggtta tgctacgata ccgatagaga tgggccaata ccacatttgt agaggtttta 5221 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt 5281 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 5341 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 5401 aatgtatctt atcatgtctg tgagctgaag gtacgctgta tctcagtcag tcaagctagc 5461 ttatttgtca tcgtcgtcct tgtaatcaat gtcatgatct ttgtagtctc catcgtggtc 5521 cttataatcc atcgactgaa agtaaaggtt ttcagttagg ttaaagttca cgacgtctac 5581 ctgagcaatg taattgcgag agttcatcct acataaaagg agtgaataga tcgtgaatgg 5641 aaaggcgaaa acatttatgt accccatggt ggcggccgct ggcgggtgtg tcgagtggat 5701 ggtaggatcg acaaagatct gatctgacgg ttcactaaac cagctctgct tatatagacc 5761 tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt 5821 ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga 5881 cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc 5941 atcagcatgg taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat 6001 aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg 6061 ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac 6121 tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt 6181 attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt 6241 tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat tgattacta //