LOCUS Exported 4497 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp7. ACCESSION . VERSION . KEYWORDS pGBW-m4134316 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4497) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4497) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4497 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 251..271 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 272..313 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" polyA_signal 366..487 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(377..396) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 431..450 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 602..648 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 678..814 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 859..930 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1156..1744) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1236..1255) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2490..3350 /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(2708..2727) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 3363..3499 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 3651..3722 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 3727..3773 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 3791..3839 /note="synthetic polyadenylation signal" enhancer 3920..4223 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4225..4427 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4378..4398 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgaaa atgtcagatg taaagtgcac atcagtagtc ttactctcag ttttgcaaca 61 actcagagta gaatcatcat ctaaattgtg ggctcaatgt gtccagttac acaatgacat 121 tctcttagct aaagatacta ctgaagcctt tgaaaaaatg gtttcactac tttctgtttt 181 gctttccatg cagggtgctg tagacataaa caagctttgt gaagaaatgc tggacaacag 241 ggcaacctta gaaaaccttt actttcagtc ggggaaacct atacccaatc cacttctcgg 301 cttagattct acctaagcta gcttgactga ctgagataca gcgtaccttc agctcacaga 361 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg 421 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa 481 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga 541 ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc 601 gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc 661 cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct 721 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc 781 atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc 841 tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga 901 agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac 961 ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc 1021 tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac 1081 gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc 1141 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg 1201 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg 1261 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt 1321 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt 1381 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg 1441 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact 1501 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt 1561 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct 1621 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac 1681 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc 1741 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg 1801 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta 1861 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca 1921 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc 1981 atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat 2041 ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct 2101 tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc 2161 gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg 2221 aatatcgaaa tcggggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga 2281 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc 2341 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta 2401 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc 2461 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta 2521 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag 2581 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca 2641 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta 2701 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc 2761 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc 2821 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca 2881 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc 2941 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca 3001 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac 3061 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg 3121 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg 3181 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg 3241 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac 3301 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag 3361 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat 3421 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt 3481 accagaaatt tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc 3541 tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat 3601 ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac 3661 cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc 3721 tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg 3781 ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta 3841 tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata 3901 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 3961 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 4021 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 4081 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 4141 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 4201 tattagtcat cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat 4261 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 4321 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc 4381 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc 4441 gtcagatcag atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc //