LOCUS Exported 6639 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp11. ACCESSION . VERSION . KEYWORDS pGBW-m4252892 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6639) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6639) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6639 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 151..222 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 227..273 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 291..339 /note="synthetic polyadenylation signal" enhancer 420..723 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 725..927 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 878..898 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 996..1005 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1038..1058 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1059..1100 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" polyA_signal 1169..1290 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1180..1199) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1234..1253 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 1405..1451 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 1481..1617 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 1662..1733 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1959..2547) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2039..2058) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 3047..4256 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 4093..4112 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 4260..5288 /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 5315..5363 /note="synthetic polyadenylation signal" CDS 5629..6489 /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(5847..5866) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 6502..6638 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" ORIGIN 1 atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg 61 cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct 121 tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct 181 tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca 241 agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa 301 accaacacac atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt 361 agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc 421 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg 481 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa 541 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca 601 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac 661 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc 721 atgctgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga 781 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg 841 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta 901 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcag atctttgtcg 961 atcctaccat ccactcgaca cacccgccag cggccgccac catggctgat gcacaatcgt 1021 ttttaaacgg gtttgcggaa aacctttact ttcagtcggg gaaacctata cccaatccac 1081 ttctcggctt agattctacc taaggatccg caggcctctg ctagcttgac tgactgagat 1141 acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca 1201 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt 1261 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt 1321 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt 1381 attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga 1441 ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat 1501 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg 1561 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt 1621 cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt 1681 tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat 1741 tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg 1801 acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc 1861 tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa 1921 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 1981 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 2041 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 2101 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 2161 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 2221 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 2281 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 2341 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga 2401 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 2461 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 2521 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 2581 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 2641 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 2701 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 2761 ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa 2821 aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa 2881 gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac 2941 tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat 3001 cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg 3061 gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac gtcagacgaa 3121 gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat 3181 aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg 3241 actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc 3301 tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg 3361 cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg 3421 tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt 3481 ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct 3541 gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc 3601 tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag 3661 gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta atgcgggaaa 3721 gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg aagtttgtca 3781 ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg 3841 ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc 3901 tgttggctta taatgcaggg tggggccacc agccggtagg tgtgcggtag gcttttctcc 3961 gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag gcgccggacc 4021 tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta tgtacctatc 4081 ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt 4141 gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt aattttcagt 4201 gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg ttagacaaca 4261 tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg 4321 acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg 4381 atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag 4441 atcgttatgt ttatcggcac tttgcatccg ccgcgctccc gattccggaa gtgcttgaca 4501 ttggggagtt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt 4561 tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggcaatgg 4621 atgccatcgc tgccgccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag 4681 gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt 4741 atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg 4801 agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg 4861 gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg 4921 cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg 4981 cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc 5041 cccggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg 5101 acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg 5161 gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgccgccgtc tggaccgatg 5221 gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa 5281 aggaataagc tagtatgtaa gcctagtctt agataataaa atcgctatcc atcgaagatg 5341 gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc gagaacgatc 5401 ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca 5461 gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt 5521 accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta 5581 gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa 5641 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac 5701 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac 5761 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt 5821 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc 5881 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca 5941 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc 6001 ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag 6061 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa 6121 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg 6181 gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag 6241 ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg 6301 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt 6361 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt 6421 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag 6481 cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc 6541 gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac 6601 ttccatacgt gtcctcctta ccagaaattt atccttaag //