LOCUS Exported 4506 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp7. ACCESSION . VERSION . KEYWORDS pGBW-m4134438 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4506) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4506) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4506 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 257..277 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 278..322 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" polyA_signal 375..496 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(386..405) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 440..459 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 611..657 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 687..823 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 868..939 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1165..1753) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1245..1264) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2499..3359 /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(2717..2736) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 3372..3508 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 3660..3731 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 3736..3782 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 3800..3848 /note="synthetic polyadenylation signal" enhancer 3929..4232 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4234..4436 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4387..4407 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgagc aaaatgtctg acgtcaagtg tacatccgta gttctgttat cagtgcttca 61 acagctgcgg gtggagagtt cgtcaaagct ctgggcccag tgcgtgcagc tgcacaatga 121 tatattgctc gcgaaagata ctacggaagc tttcgagaag atggtcagct tgctatctgt 181 tctactgagc atgcaaggag cagtagacat caacaaactt tgcgaagaga tgttagacaa 241 ccgagccacc ctgcaagaaa acctttactt tcagtcgaaa gaaaccgcgg cagctaagtt 301 cgagaggcag cacatggaca gttaagctag cttgactgac tgagatacag cgtaccttca 361 gctcacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg 421 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag 481 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga 541 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg gcccatctct 601 atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgtgc 661 ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc 721 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg 781 cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc agctatcctg 841 caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg gacaccacta 901 ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt taccttgggc 961 tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt 1021 cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc 1081 tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg 1141 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 1201 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 1261 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 1321 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 1381 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 1441 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 1501 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 1561 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 1621 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 1681 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 1741 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga 1801 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct 1861 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga 1921 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 1981 catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt 2041 tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc 2101 cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata 2161 tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga 2221 gatccgagga atatcgaaat cggggcgcgc ctggtgtacc gagaacgatc ctctcagtgc 2281 gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc 2341 caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt 2401 ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa 2461 acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa catttccgtg 2521 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc 2581 tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac atcgaactgg 2641 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt ccaatgatga 2701 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc 2761 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag 2821 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga 2881 gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg 2941 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga 3001 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct 3061 tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag ttgatagact 3121 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt 3181 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg 3241 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta 3301 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac 3361 cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact 3421 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt 3481 gtcctcctta ccagaaattt atccttaaga tcccgaatcg tttaaactcg actctggctc 3541 tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg 3601 gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg cagcgatcgc ggctcccgac 3661 atcttggacc attagctcca caggtatctt cttccctcta gtggtcataa cagcagcttc 3721 agctacctct caattcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc 3781 tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca tccatcttcg atggatagcg 3841 attttattat ctaactgctg atcgagtgta gccagatcta gtaatcaatt acggggtcat 3901 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg 3961 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa 4021 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact 4081 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta 4141 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt 4201 acatctacgt attagtcatc gctattacca tgctgatgcg gttttggcag tacatcaatg 4261 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg 4321 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc 4381 cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt 4441 tagtgaaccg tcagatcaga tctttgtcga tcctaccatc cactcgacac acccgccagc 4501 ggccgc //