LOCUS Exported 6236 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp11. ACCESSION . VERSION . KEYWORDS pGBW-m4252799 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6236) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6236) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6236 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 92..140 /note="synthetic polyadenylation signal" terminator 158..204 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" misc_feature 209..280 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 432..568 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" CDS complement(581..1441) /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 1204..1223 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" polyA_signal 1707..1755 /note="synthetic polyadenylation signal" CDS complement(1782..2381) /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(1865..1885) /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind 2362..2381 /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" promoter complement(2385..3594) /label=UbC promoter /note="human ubiquitin C promoter" primer_bind complement(2529..2548) /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" rep_origin 4094..4682 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4583..4602 /label=pBR322ori-F /note="pBR322 origin, forward primer" misc_feature 4908..4979 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" terminator 5024..5160 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" terminator 5190..5236 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 5351..5472 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5388..5407) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5442..5461 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" CDS complement(5525..5602) /codon_start=1 /product="calmodulin-binding peptide" /label=CBP /note="derived from skeletal muscle myosin light chain kinase; binds calmodulin with nanomolar affinity in the presence of calcium" /translation="KRRWKKNFIAVSAANRFKKISSSGAL" CDS complement(5603..5623) /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" promoter complement(5740..5942) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(5769..5789) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 5944..11 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact 61 agatctggct acactcgatc agcagttaga taataaaatc gctatccatc gaagatggat 121 gtgtgttggt tttttgtgtg tgtaacgcaa cgattgatag cataacccct tggggcctct 181 aaacgggtct tgaggggttt tttgaattga gaggtagctg aagctgctgt tatgaccact 241 agagggaaga agatacctgt ggagctaatg gtccaagatg tcgggagccg cgatcgctgc 301 caaaaaggta agctgacgat agctgacgag attcgatgcc cgacgagcaa atgagcgcca 361 cggctgttcg cgtaagctcg aaacgacgga gattcgatag agccagagtc gagtttaaac 421 gattcgggat cttaaggata aatttctggt aaggaggaca cgtatggaag tgggcaagtt 481 ggggaagccg tatccgttgc tgaatctggc atatgtggga gtataagacg cgcagcgtcg 541 catcaggcat ttttttctgc gccaatgcac ctagaatcgg ttaccaatgc ttaatcagtg 601 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 661 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 721 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 781 agcgcagaag tggtcctgca actttatccg cctccatcca gtctatcaac tgttgccggg 841 aagctagagt aagtagttcg ccagttaata gtttacgcaa ggttgttgcc attgctacag 901 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 961 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 1021 caatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 1081 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gaatactcaa 1141 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 1201 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aagcgttctt 1261 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 1321 gcgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 1381 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 1441 tgcgaaagcc tcctgctgat cctgtctctt gatagatgtt tgcaaaagct aggactcgat 1501 tatacgttga ccgatcagac tatcaatatc taggtttaaa cagatcggta cgctgccgtg 1561 accaggcttg agtacaatat ctgacgattg gacttcctgg gtcccaagct ggattccgac 1621 tggctgactg ccaagcaacg atatggatcg tcgagactcg cactgagagg atcgttctcg 1681 gtacaccagg cgcgcctagc gttacacaca caaaaaacca acacacatcc atcttcgatg 1741 gatagcgatt ttattatcta agactaggct tacatactag cctaggcacc gggcttgcgg 1801 gtcatgcacc aggtgcgcgg tccttcgggc acctcgacgt cggcggtgac ggtgaagccg 1861 agccgctcgt agaaggggag gttgcggggc gcggaggtct ccaggaaggc gggcaccccg 1921 gcgcgctcgg cggcctccac tccggggagc acgacggcgc tgcccagacc cttgccctgg 1981 tggtcgggcg agacgccgac ggtggccagg aaccacgcgg gctccttggg ccggtgcggc 2041 gccaggaggc cctccatctg ttgctgcgcg gccagccggg aaccgctcaa ctcggccatg 2101 cgcgggccga tctcggcgaa caccgccccc gcttcgacgc tctccggcgt ggtccagacc 2161 gccaccgcgg cgccgtcgtc cgcgacccac accttgccga tgtcgagccc gacgcgcgtg 2221 aggaagagtt cttgcagctc ggtgacccgc tcgatgtggc ggtccggatc gacggtgtgg 2281 cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc gtacggccct ggggacgtcg 2341 tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca tgttgtctaa caaaaaagcc 2401 aaaaacggcc agaatttagc ggacaattta caagtctaac actgaaaatt acatattgac 2461 ccaaatgatt acatttcaaa aggtgcctaa aaaacttcac aaaacacact cgccaacccc 2521 gagcgcatag ttcaaaaccg gagcttcagc tacttaagaa gataggtaca taaaaccgac 2581 caaagaaact gacgcctcac ttatccctcc cctcaccaga ggtccggcgc ctgtcgattc 2641 aggagagcct accctaggcc cgaaccctgc gtcctgcgac ggagaaaagc ctaccgcaca 2701 cctaccggct ggtggcccca ccctgcatta taagccaaca gaacgggtga cgtcacgaca 2761 cgacgagggc gcgcgctccc aaaggtacgg gtgcactgcc caacggcacc gccataactg 2821 ccgcccccgc aacagacgac aaaccgagtt ctccagtcag tgacaaactt cacgtcaggg 2881 tccccagatg gtgcccagcc catctcaccc gaataagagc tttcccgcat tagcgaaggg 2941 ctcaagacct tgggttcttg ccgcccacca tgccccccac cttgtttcaa cgacctcaca 3001 gcccgcctca caagcgtctt ccattcaaga ctcgggaaca gccgccattt tgctgcgctc 3061 cccccaaccc ccagttcagg gcaaccttgc tcgcggaccc agactacagc ccttggcggt 3121 ctctccacac gcttccgtcc caccgagcgg cccggcggcc acgaaagccc cggccagccc 3181 agcagcccgc tactcaccaa gtgacgatca cagcgatcca caaacaagaa ccgcgaccca 3241 aatcccggct gcgacggaac tagctgtgcc acacccggcg cgtccttata taatcatcgg 3301 cgttcaccgc cccacggaga tccctccgca gaatcgccga gaagggacta cttttcctcg 3361 cctgttccgc tctctggaaa gaaaaccagt gccctagagt cacccaagtc ccgtcctaaa 3421 atgtccttct gctgatactg gggttctaag gccgagtctt atgagcagcg ggccgctgtc 3481 ctgagcgtcc gggcggaagg atcaggacgc tcctgcgccc ttcgtctgac gtggcagcgc 3541 tcgccgtgac gaggggggcg cccgcgggag gcgccaaaac ccggcgcgga ggccaggcgc 3601 gccccgattt cgatattcct cggatctcgg atagatcggg attgaggatg taggttgaac 3661 ttctctcggg tgatcccgat ctcggatatt cgggattgga gtaatacctg gcgataggcg 3721 ctgccaagca atagccaagg cgcaaggccg gccaagagac ttgcgatagt tcgttcgaga 3781 ggtagcctgc aagatggatc ggacgaaagg tttgaagatt ttccgacgag ggccttggag 3841 agttcggaaa tttaaatgca actatggatg aacgaaatag acagatcgct gagataggtg 3901 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg 3961 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca 4021 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga 4081 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 4141 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 4201 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt 4261 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 4321 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 4381 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 4441 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 4501 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 4561 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 4621 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 4681 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 4741 tgatttaaat gaaactcgta atcggagaac gggtgcaata gcaaggacca gatcgaactt 4801 gactgacgca attctgagcg tagtcgaaac ccagacaagt cgatctcgcc aagtagctga 4861 cttgactggt taattaagtt tcgatagccc aaggtaacca atgagattag ggagggaaga 4921 aagtttgatg aactacttct gacccctagt ggtgtccaga aaagaccatt aaaggaatgt 4981 cttgatcgaa atcgagagat cgcctgcagg atagctgacg accttaagga taaatttctg 5041 gtaaggagga cacgtatgga agtgggcaag ttggggaagc cgtatccgtt gctgaatctg 5101 gcatatgtgg gagtataaga cgcgcagcgt cgcatcaggc atttttttct gcgccaatgc 5161 cggtagatag caatccggcc cgaggggcac aaaaaacccc tcaagacccg tttagaggcc 5221 ccaaggggtt atgctacgat accgatagag atgggccaat accacatttg tagaggtttt 5281 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat 5341 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 5401 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 5461 caatgtatct tatcatgtct gtgagctgaa ggtacgctgt atctcagtca gtcaagctag 5521 cttagagggc accagacgag gatatctttt tgaatctgtt ggctgcactc accgcgataa 5581 aattcttttt ccatcgcctc ttcgactgaa agtaaaggtt ttccactgcg aaaccgttga 5641 gaaaggactg agcatcggca ctcatggtgg cggccgctgg cgggtgtgtc gagtggatgg 5701 taggatcgac aaagatctga tctgacggtt cactaaacca gctctgctta tatagacctc 5761 ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg 5821 aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact 5881 tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat 5941 cagcatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa 6001 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg 6061 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc 6121 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat 6181 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaa //