LOCUS Exported 5637 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp6. ACCESSION . VERSION . KEYWORDS pGBW-m4134163 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5637) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5637) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5637 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 878..898 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 932..1009 /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 1037..1057 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" polyA_signal 1506..1627 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1517..1536) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1571..1590 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 1742..1788 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 1818..1954 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 1999..2070 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(2296..2884) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2376..2395) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 3630..4490 /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(3848..3867) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 4503..4639 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 4791..4862 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 4867..4913 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 4931..4979 /note="synthetic polyadenylation signal" enhancer 5060..5363 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 5365..5567 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5518..5538 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgagc gccgttaagc ggacaatcaa agggacccat cactggttgc tccttactat 61 actgacgtct ttactggtct tggtacagag tacacaatgg tcgcttttct tttttctgta 121 cgagaatgca ttcctcccat ttgcgatggg aatcattgct atgtccgcct tcgctatgat 181 gttcgtgaag cacaaacatg ccttcctttg cctgtttctc ctaccctcac tggcaaccgt 241 ggcctatttt aacatggtgt acatgcctgc gagctgggtc atgcgaatta tgacttggct 301 ggacatggtt gatacctccc tatctggctt caagttaaag gactgtgtga tgtatgcatc 361 agctgtagtg ctgctaatcc tgatgacagc tcgcacagtc tacgatgacg gtgccaggag 421 agtgtggact ctgatgaatg tcctcacctt agtttataaa gtatactatg gcaacgctct 481 cgatcaggcc attagtatgt gggcattgat aatctccgtg acgagtaact acagcggagt 541 ggtaaccact gtcatgtttt tggcacgcgg gatagtgttc atgtgtgtcg aatattgccc 601 gatctttttt atcacaggga atactctgca atgcattatg ctggtttact gtttcttggg 661 ctatttctgc acctgttatt ttggtctatt ctgcctgctt aataggtact ttcgtttgac 721 actaggagtc tatgattact tggtgagcac gcaggagttt cggtatatga acagtcaggg 781 cctgcttccc ccaaagaact ctattgacgc gtttaaactc aatataaagc tgttgggcgt 841 gggtggaaaa ccttgcatca aggtggccac tgtgcaggaa aacctttact ttcagtcggg 901 acggcgaatc ccggggctga taaatccctg gaaaaggaga tggaagaaga actttattgc 961 agtgtctgcg gctaaccggt tcaaaaagat cagttcaagc ggcgccctag attacgacat 1021 tcctacgacc gcctccgaga atctctattt ccagggtgaa tttggcttgg cacaacatga 1081 cgaggccgtc gataacaagt ttaataaaga acagcagaac gctttctatg agatccttca 1141 cttaccaaat ctgaacgaag agcaacgcaa cgctttcatt cagagcctga aagacgatcc 1201 ctcgcagtct gcaaatctcc tggccgaagc caagaagctg aatgacgcac aggctccaaa 1261 agttgataac aagtttaaca aggagcagca gaatgctttc tacgagatat tgcaccttcc 1321 taatctgaat gaggaacaaa gaaatgcatt tatccagtcc ctaaaagacg atccatcaca 1381 gagtgccaac ctgcttgcgg aagccaaaaa gctcaatgat gcccaggccc cgaaagtaga 1441 cgcgaaccat caataagcta gcttgactga ctgagataca gcgtaccttc agctcacaga 1501 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg 1561 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa 1621 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga 1681 ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc 1741 gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc 1801 cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct 1861 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc 1921 atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc 1981 tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga 2041 agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac 2101 ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc 2161 tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac 2221 gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc 2281 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg 2341 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg 2401 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt 2461 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt 2521 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg 2581 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact 2641 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt 2701 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct 2761 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac 2821 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc 2881 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg 2941 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta 3001 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca 3061 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc 3121 atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat 3181 ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct 3241 tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc 3301 gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg 3361 aatatcgaaa tcggggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga 3421 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc 3481 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta 3541 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc 3601 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta 3661 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag 3721 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca 3781 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta 3841 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc 3901 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc 3961 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca 4021 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc 4081 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca 4141 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac 4201 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg 4261 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg 4321 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg 4381 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac 4441 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag 4501 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat 4561 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt 4621 accagaaatt tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc 4681 tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat 4741 ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac 4801 cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc 4861 tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg 4921 ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta 4981 tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata 5041 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 5101 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 5161 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 5221 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 5281 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 5341 tattagtcat cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat 5401 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 5461 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc 5521 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc 5581 gtcagatcag atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc //