LOCUS Exported 6576 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp11. ACCESSION . VERSION . KEYWORDS pGBW-m4252982 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6576) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6576) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6576 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 670..1879 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 1716..1735 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 1883..2911 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA KE" polyA_signal 2938..2986 /note="synthetic polyadenylation signal" CDS 3252..4112 /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(3470..3489) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 4125..4261 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 4413..4484 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 4489..4535 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 4553..4601 /note="synthetic polyadenylation signal" enhancer 4682..4985 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4987..5189 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5140..5160 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 5258..5267 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" polyA_signal 5368..5489 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5379..5398) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 5433..5452 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 5604..5650 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 5680..5816 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 5861..5932 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(6158..170) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6238..6257) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 61 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 121 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 181 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 241 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 301 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 361 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga 421 actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct 481 acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg 541 cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga 601 gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg 661 ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg 721 cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg cccggacgct 781 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga 841 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac 901 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga 961 acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg 1021 atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct 1081 gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga 1141 gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg 1201 ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg 1261 ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt 1321 gagcccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg caccatctgg 1381 ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg 1441 cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg 1501 tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc accagccggt 1561 aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct 1621 cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct 1681 ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg 1741 ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt 1801 tgggtcaata tgtaattttc agtgttagac ttgtaaattg tccgctaaat tctggccgtt 1861 tttggctttt ttgttagaca acatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga 1921 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga 1981 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag 2041 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat ccgccgcgct 2101 cccgattccg gaagtgcttg acattgggga gttcagcgag agcctgacct attgcatctc 2161 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct 2221 gcagccggtc gcggaggcaa tggatgccat cgctgccgcc gatcttagcc agacgagcgg 2281 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg 2341 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc 2401 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg 2461 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac 2521 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat 2581 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag 2641 gcatccggag cttgcaggat cgccccggct ccgggcgtat atgctccgca ttggtcttga 2701 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg 2761 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag 2821 aagcgccgcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg 2881 ccccagcact cgtccgaggg caaaggaata agctagtatg taagcctagt cttagataat 2941 aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg 3001 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt 3061 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta 3121 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc 3181 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag 3241 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 3301 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 3361 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 3421 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg 3481 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 3541 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag 3601 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 3661 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 3721 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 3781 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac 3841 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac 3901 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 3961 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 4021 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 4081 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa 4141 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga 4201 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta 4261 agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta 4321 cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag 4381 cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat 4441 cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct 4501 caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa 4561 aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt 4621 gtagccagat ctagtaatca attacggggt cattagttca tagcccatat atggagttcc 4681 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat 4741 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc 4801 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc 4861 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt 4921 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta 4981 ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg 5041 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac 5101 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg 5161 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt 5221 cgatcctacc atccactcga cacacccgcc agcggccgcc accatggctg atgcacaatc 5281 gtttttaaac gggtttgcgt aaggatccgc aggcctctgc tagcttgact gactgagata 5341 cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa 5401 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg 5461 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc 5521 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta 5581 ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag 5641 gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg 5701 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg 5761 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc 5821 gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt 5881 ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt 5941 ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga 6001 cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct 6061 attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa 6121 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg 6181 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa 6241 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc 6301 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac 6361 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac 6421 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg 6481 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt 6541 atgtaggcgg tgctacagag ttcttgaagt ggtggc //