LOCUS Exported 6131 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp11. ACCESSION . VERSION . KEYWORDS pGBW-m4252976 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6131) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6131) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6131 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 96..144 /note="synthetic polyadenylation signal" CDS complement(171..770) /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(254..274) /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind 751..770 /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" promoter complement(774..1983) /label=UbC promoter /note="human ubiquitin C promoter" primer_bind complement(918..937) /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" rep_origin 2483..3071 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2972..2991 /label=pBR322ori-F /note="pBR322 origin, forward primer" misc_feature 3297..3368 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" terminator 3413..3549 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" terminator 3579..3625 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 3740..3861 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3777..3796) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3831..3850 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" regulatory 3946..3955 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter complement(4024..4226) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(4053..4073) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 4228..4531 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" polyA_signal 4612..4660 /note="synthetic polyadenylation signal" terminator 4678..4724 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" misc_feature 4729..4800 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 4952..5088 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" CDS complement(5101..5961) /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 5724..5743 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 gattccgact ggctgactgc caagcaacga tatggatcgt cgagactcgc actgagagga 61 tcgttctcgg tacaccaggc gcgcctagcg ttacacacac aaaaaaccaa cacacatcca 121 tcttcgatgg atagcgattt tattatctaa gactaggctt acatactagc ctaggcaccg 181 ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg 241 gtgaagccga gccgctcgta gaaggggagg ttgcggggcg cggaggtctc caggaaggcg 301 ggcaccccgg cgcgctcggc ggcctccact ccggggagca cgacggcgct gcccagaccc 361 ttgccctggt ggtcgggcga gacgccgacg gtggccagga accacgcggg ctccttgggc 421 cggtgcggcg ccaggaggcc ctccatctgt tgctgcgcgg ccagccggga accgctcaac 481 tcggccatgc gcgggccgat ctcggcgaac accgcccccg cttcgacgct ctccggcgtg 541 gtccagaccg ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg 601 acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg 661 acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg 721 gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat gttgtctaac 781 aaaaaagcca aaaacggcca gaatttagcg gacaatttac aagtctaaca ctgaaaatta 841 catattgacc caaatgatta catttcaaaa ggtgcctaaa aaacttcaca aaacacactc 901 gccaaccccg agcgcatagt tcaaaaccgg agcttcagct acttaagaag ataggtacat 961 aaaaccgacc aaagaaactg acgcctcact tatccctccc ctcaccagag gtccggcgcc 1021 tgtcgattca ggagagccta ccctaggccc gaaccctgcg tcctgcgacg gagaaaagcc 1081 taccgcacac ctaccggctg gtggccccac cctgcattat aagccaacag aacgggtgac 1141 gtcacgacac gacgagggcg cgcgctccca aaggtacggg tgcactgccc aacggcaccg 1201 ccataactgc cgcccccgca acagacgaca aaccgagttc tccagtcagt gacaaacttc 1261 acgtcagggt ccccagatgg tgcccagccc atctcacccg aataagagct ttcccgcatt 1321 agcgaagggc tcaagacctt gggttcttgc cgcccaccat gccccccacc ttgtttcaac 1381 gacctcacag cccgcctcac aagcgtcttc cattcaagac tcgggaacag ccgccatttt 1441 gctgcgctcc ccccaacccc cagttcaggg caaccttgct cgcggaccca gactacagcc 1501 cttggcggtc tctccacacg cttccgtccc accgagcggc ccggcggcca cgaaagcccc 1561 ggccagccca gcagcccgct actcaccaag tgacgatcac agcgatccac aaacaagaac 1621 cgcgacccaa atcccggctg cgacggaact agctgtgcca cacccggcgc gtccttatat 1681 aatcatcggc gttcaccgcc ccacggagat ccctccgcag aatcgccgag aagggactac 1741 ttttcctcgc ctgttccgct ctctggaaag aaaaccagtg ccctagagtc acccaagtcc 1801 cgtcctaaaa tgtccttctg ctgatactgg ggttctaagg ccgagtctta tgagcagcgg 1861 gccgctgtcc tgagcgtccg ggcggaagga tcaggacgct cctgcgccct tcgtctgacg 1921 tggcagcgct cgccgtgacg aggggggcgc ccgcgggagg cgccaaaacc cggcgcggag 1981 gccaggcgcg ccccgatttc gatattcctc ggatctcgga tagatcggga ttgaggatgt 2041 aggttgaact tctctcgggt gatcccgatc tcggatattc gggattggag taatacctgg 2101 cgataggcgc tgccaagcaa tagccaaggc gcaaggccgg ccaagagact tgcgatagtt 2161 cgttcgagag gtagcctgca agatggatcg gacgaaaggt ttgaagattt tccgacgagg 2221 gccttggaga gttcggaaat ttaaatgcaa ctatggatga acgaaataga cagatcgctg 2281 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 2341 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 2401 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 2461 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 2521 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 2581 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt 2641 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 2701 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 2761 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 2821 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 2881 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 2941 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 3001 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 3061 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 3121 ttgctcacat gatttaaatg aaactcgtaa tcggagaacg ggtgcaatag caaggaccag 3181 atcgaacttg actgacgcaa ttctgagcgt agtcgaaacc cagacaagtc gatctcgcca 3241 agtagctgac ttgactggtt aattaagttt cgatagccca aggtaaccaa tgagattagg 3301 gagggaagaa agtttgatga actacttctg acccctagtg gtgtccagaa aagaccatta 3361 aaggaatgtc ttgatcgaaa tcgagagatc gcctgcagga tagctgacga ccttaaggat 3421 aaatttctgg taaggaggac acgtatggaa gtgggcaagt tggggaagcc gtatccgttg 3481 ctgaatctgg catatgtggg agtataagac gcgcagcgtc gcatcaggca tttttttctg 3541 cgccaatgcc ggtagatagc aatccggccc gaggggcaca aaaaacccct caagacccgt 3601 ttagaggccc caaggggtta tgctacgata ccgatagaga tgggccaata ccacatttgt 3661 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat 3721 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa 3781 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 3841 caaactcatc aatgtatctt atcatgtctg tgagctgaag gtacgctgta tctcagtcag 3901 tcaagctagc ttacgcaaac ccgtttaaaa acgattgtgc atcagccatg gtggcggccg 3961 ctggcgggtg tgtcgagtgg atggtaggat cgacaaagat ctgatctgac ggttcactaa 4021 accagctctg cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg 4081 ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc 4141 attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc 4201 attgatgtac tgccaaaacc gcatcagcat ggtaatagcg atgactaata cgtagatgta 4261 ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt 4321 accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa 4381 gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt 4441 tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc 4501 aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa 4561 tgaccccgta attgattact agatctggct acactcgatc agcagttaga taataaaatc 4621 gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcaa cgattgatag 4681 cataacccct tggggcctct aaacgggtct tgaggggttt tttgaattga gaggtagctg 4741 aagctgctgt tatgaccact agagggaaga agatacctgt ggagctaatg gtccaagatg 4801 tcgggagccg cgatcgctgc caaaaaggta agctgacgat agctgacgag attcgatgcc 4861 cgacgagcaa atgagcgcca cggctgttcg cgtaagctcg aaacgacgga gattcgatag 4921 agccagagtc gagtttaaac gattcgggat cttaaggata aatttctggt aaggaggaca 4981 cgtatggaag tgggcaagtt ggggaagccg tatccgttgc tgaatctggc atatgtggga 5041 gtataagacg cgcagcgtcg catcaggcat ttttttctgc gccaatgcac ctagaatcgg 5101 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 5161 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 5221 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 5281 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 5341 gtctatcaac tgttgccggg aagctagagt aagtagttcg ccagttaata gtttacgcaa 5401 ggttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 5461 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 5521 ggttagctcc ttcggtcctc caatcgttgt cagaagtaag ttggccgcag tgttatcact 5581 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 5641 tgtgactggt gaatactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 5701 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 5761 catcattgga aagcgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 5821 cagttcgatg taacccactc gcgcacccaa ctgatcttca gcatctttta ctttcaccag 5881 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 5941 acggaaatgt tgaatactca tgcgaaagcc tcctgctgat cctgtctctt gatagatgtt 6001 tgcaaaagct aggactcgat tatacgttga ccgatcagac tatcaatatc taggtttaaa 6061 cagatcggta cgctgccgtg accaggcttg agtacaatat ctgacgattg gacttcctgg 6121 gtcccaagct g //