LOCUS Exported 5061 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 ORF3a protein. ACCESSION . VERSION . KEYWORDS pGBW-m4133717 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5061) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5061) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5061 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..304 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 306..508 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 459..479 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 577..586 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1408..1428 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1429..1455 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 1508..1629 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1519..1538) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1573..1592 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 1744..1790 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 1820..1956 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 2001..2072 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(2298..2886) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2378..2397) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 3632..4492 /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(3850..3869) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 4505..4641 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 4793..4864 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 4869..4915 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 4933..4981 /note="synthetic polyadenylation signal" ORIGIN 1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc 541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatggattt gtttatgaga 601 atcttcacaa ttggaactgt aactttgaag caaggtgaaa tcaaggatgc tactccttca 661 gattttgttc gcgctactgc aacgataccg atacaagcct cactcccttt cggatggctt 721 attgttggcg ttgcacttct tgctgttttt cagagcgctt ccaaaatcat aaccctcaaa 781 aagagatggc aactagcact ctccaagggt gttcactttg tttgcaactt gctgttgttg 841 tttgtaacag tttactcaca ccttttgctc gttgctgctg gccttgaagc cccttttctc 901 tatctttatg ctttagtcta cttcttgcag agtataaact ttgtaagaat aataatgagg 961 ctttggcttt gctggaaatg ccgttccaaa aacccattac tttatgatgc caactatttt 1021 ctttgctggc atactaattg ttacgactat tgtatacctt acaatagtgt aacttcttca 1081 attgtcatta cttcaggtga tggcacaaca agtcctattt ctgaacatga ctaccagatt 1141 ggtggttata ctgaaaaatg ggaatctgga gtaaaagact gtgttgtatt acacagttac 1201 ttcacttcag actattacca gctgtactca actcaattga gtacagacac tggtgttgaa 1261 catgttacct tcttcatcta caataaaatt gttgatgagc ctgaagaaca tgtccaaatt 1321 cacacaatcg acggttcatc cggagttgtt aatccagtaa tggaaccaat ttatgatgaa 1381 ccgacgacga ctactagcgt gcctttggaa aacctttact ttcagtcgta cccctatgac 1441 gtaccagatt atgcgtaagc tagcttgact gactgagata cagcgtacct tcagctcaca 1501 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa 1561 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat 1621 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg 1681 gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc tctatcggta 1741 tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg tgcccctcgg 1801 gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt 1861 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt 1921 ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc ctgcaggcga 1981 tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca ctaggggtca 2041 gaagtagttc atcaaacttt cttccctccc taatctcatt ggttaccttg ggctatcgaa 2101 acttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg tttcgactac 2161 gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg ttctccgatt 2221 acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 2281 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 2341 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 2401 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 2461 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 2521 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 2581 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 2641 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 2701 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 2761 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 2821 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 2881 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 2941 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 3001 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 3061 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 3121 gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg 3181 atccatcttg caggctacct ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc 3241 cttggctatt gcttggcagc gcctatcgcc aggtattact ccaatcccga atatccgaga 3301 tcgggatcac ccgagagaag ttcaacctac atcctcaatc ccgatctatc cgagatccga 3361 ggaatatcga aatcggggcg cgcctggtgt accgagaacg atcctctcag tgcgagtctc 3421 gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg acccaggaag 3481 tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc 3541 tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg caaacatcta 3601 tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc gtgtcgccct 3661 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa 3721 agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac tggatctcaa 3781 cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga tgagcacttt 3841 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg 3901 tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca cagaaaagca 3961 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa 4021 cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa ccgctttttt 4081 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc 4141 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa ccttgcgtaa 4201 actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag actggatgga 4261 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc 4321 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga 4381 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga 4441 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aaccgattct 4501 aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat 4561 atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc 4621 ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa 4681 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga 4741 atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg 4801 accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc 4861 tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat 4921 cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat 4981 tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca 5041 tagcccatat atggagttcc g //