LOCUS Exported 6858 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp7. ACCESSION . VERSION . KEYWORDS pGBW-m4252881 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6858) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6858) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6858 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 221..809 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 710..729 /label=pBR322ori-F /note="pBR322 origin, forward primer" misc_feature 1035..1106 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" terminator 1151..1287 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" terminator 1317..1363 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 1478..1599 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1515..1534) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1569..1588 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" CDS complement(1668..1712) /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS complement(1713..1733) /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" promoter complement(2060..2262) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(2089..2109) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 2264..2567 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" polyA_signal 2648..2696 /note="synthetic polyadenylation signal" terminator 2714..2760 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" misc_feature 2765..2836 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 2988..3124 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" CDS complement(3137..3997) /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 3760..3779 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" polyA_signal 4263..4311 /note="synthetic polyadenylation signal" CDS complement(4338..5366) /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" promoter complement(5370..6579) /label=UbC promoter /note="human ubiquitin C promoter" primer_bind complement(5514..5533) /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" ORIGIN 1 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc 61 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct 121 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc 181 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 241 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 301 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 361 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 421 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt 481 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa 541 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 601 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc 661 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct 721 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat 781 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc 841 tggccttttg ctggcctttt gctcacatga tttaaatgaa actcgtaatc ggagaacggg 901 tgcaatagca aggaccagat cgaacttgac tgacgcaatt ctgagcgtag tcgaaaccca 961 gacaagtcga tctcgccaag tagctgactt gactggttaa ttaagtttcg atagcccaag 1021 gtaaccaatg agattaggga gggaagaaag tttgatgaac tacttctgac ccctagtggt 1081 gtccagaaaa gaccattaaa ggaatgtctt gatcgaaatc gagagatcgc ctgcaggata 1141 gctgacgacc ttaaggataa atttctggta aggaggacac gtatggaagt gggcaagttg 1201 gggaagccgt atccgttgct gaatctggca tatgtgggag tataagacgc gcagcgtcgc 1261 atcaggcatt tttttctgcg ccaatgccgg tagatagcaa tccggcccga ggggcacaaa 1321 aaacccctca agacccgttt agaggcccca aggggttatg ctacgatacc gatagagatg 1381 ggccaatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct 1441 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa 1501 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 1561 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgtg agctgaaggt 1621 acgctgtatc tcagtcagtc aagctagcag aggcctgcgg atccttaact gtccatgtgc 1681 tgcctctcga acttagctgc cgcggtttct ttcgactgaa agtaaaggtt ttcttgcagg 1741 gtggctcggt tgtctaacat ctcttcgcaa agtttgttga tgtctactgc tccttgcatg 1801 ctcagtagaa cagatagcaa gctgaccatc ttctcgaaag cttccgtagt atctttcgcg 1861 agcaatatat cattgtgcag ctgcacgcac tgggcccaga gctttgacga actctccacc 1921 cgcagctgtt gaagcactga taacagaact acggatgtac acttgacgtc agacattttg 1981 ctcatggtgg cggccgctgg cgggtgtgtc gagtggatgg taggatcgac aaagatctga 2041 tctgacggtt cactaaacca gctctgctta tatagacctc ccaccgtaca cgcctaccgc 2101 ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt 2161 gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa 2221 accgctatcc acgcccattg atgtactgcc aaaaccgcat cagcatggta atagcgatga 2281 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat 2341 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca 2401 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga 2461 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt 2521 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat 2581 atgggctatg aactaatgac cccgtaattg attactagat ctggctacac tcgatcagca 2641 gttagataat aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta 2701 acgcaacgat tgatagcata accccttggg gcctctaaac gggtcttgag gggttttttg 2761 aattgagagg tagctgaagc tgctgttatg accactagag ggaagaagat acctgtggag 2821 ctaatggtcc aagatgtcgg gagccgcgat cgctgccaaa aaggtaagct gacgatagct 2881 gacgagattc gatgcccgac gagcaaatga gcgccacggc tgttcgcgta agctcgaaac 2941 gacggagatt cgatagagcc agagtcgagt ttaaacgatt cgggatctta aggataaatt 3001 tctggtaagg aggacacgta tggaagtggg caagttgggg aagccgtatc cgttgctgaa 3061 tctggcatat gtgggagtat aagacgcgca gcgtcgcatc aggcattttt ttctgcgcca 3121 atgcacctag aatcggttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 3181 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 3241 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 3301 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 3361 tatccgcctc catccagtct atcaactgtt gccgggaagc tagagtaagt agttcgccag 3421 ttaatagttt acgcaaggtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt 3481 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca 3541 tgttgtgcaa aaaagcggtt agctccttcg gtcctccaat cgttgtcaga agtaagttgg 3601 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat 3661 ccgtaagatg cttttctgtg actggtgaat actcaaccaa gtcattctga gaatagtgta 3721 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca 3781 gaactttaaa agtgctcatc attggaaagc gttcttcggg gcgaaaactc tcaaggatct 3841 taccgctgtt gagatccagt tcgatgtaac ccactcgcgc acccaactga tcttcagcat 3901 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa 3961 agggaataag ggcgacacgg aaatgttgaa tactcatgcg aaagcctcct gctgatcctg 4021 tctcttgata gatgtttgca aaagctagga ctcgattata cgttgaccga tcagactatc 4081 aatatctagg tttaaacaga tcggtacgct gccgtgacca ggcttgagta caatatctga 4141 cgattggact tcctgggtcc caagctggat tccgactggc tgactgccaa gcaacgatat 4201 ggatcgtcga gactcgcact gagaggatcg ttctcggtac accaggcgcg cctagcgtta 4261 cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat tatctaagac 4321 taggcttaca tactagctta ttcctttgcc ctcggacgag tgctggggcg tcggtttcca 4381 ctatcggcga gtacttctac acagccatcg gtccagacgg cggcgcttct gcgggcgatt 4441 tgtgtacgcc cgacagtccc ggctccggat cggacgattg cgtcgcatcg accctgcgcc 4501 caagctgcat catcgaaatt gccgtcaacc aagctctgat agagttggtc aagaccaatg 4561 cggagcatat acgcccggag ccggggcgat cctgcaagct ccggatgcct ccgctcgaag 4621 tagcgcgtct gctgctccat acaagccaac cacggcctcc agaagaagat gttggcgacc 4681 tcgtattggg aatccccgaa catcgcctcg ctccagtcaa tgaccgctgt tatgcggcca 4741 ttgtccgtca ggacattgtt ggagccgaaa tccgcgtgca cgaggtgccg gacttcgggg 4801 cagtcctcgg cccaaagcat cagctcatcg agagcctgcg cgacggacgc actgacggtg 4861 tcgtccatca cagtttgcca gtgatacaca tggggatcag caatcgcgca tatgaaatca 4921 cgccatgtag tgtattgacc gattccttgc ggtccgaatg ggccgaaccc gctcgtctgg 4981 ctaagatcgg cggcagcgat ggcatccatt gcctccgcga ccggctgcag aacagcgggc 5041 agttcggttt caggcaggtc ttgcaacgtg acaccctgtg cacggcggga gatgcaatag 5101 gtcaggctct cgctgaactc cccaatgtca agcacttccg gaatcgggag cgcggcggat 5161 gcaaagtgcc gataaacata acgatctttg tagaaaccat cggcgcagct atttacccgc 5221 aggacatatc cacgccctcc tacatcgaag ctgaaagcac gagattcttc gccctccgag 5281 agctgcatca ggtcggagac gctgtcgaac ttttcgatca gaaacttctc gacagacgtc 5341 gcggtgagtt caggcttttt acccatgttg tctaacaaaa aagccaaaaa cggccagaat 5401 ttagcggaca atttacaagt ctaacactga aaattacata ttgacccaaa tgattacatt 5461 tcaaaaggtg cctaaaaaac ttcacaaaac acactcgcca accccgagcg catagttcaa 5521 aaccggagct tcagctactt aagaagatag gtacataaaa ccgaccaaag aaactgacgc 5581 ctcacttatc cctcccctca ccagaggtcc ggcgcctgtc gattcaggag agcctaccct 5641 aggcccgaac cctgcgtcct gcgacggaga aaagcctacc gcacacctac cggctggtgg 5701 ccccaccctg cattataagc caacagaacg ggtgacgtca cgacacgacg agggcgcgcg 5761 ctcccaaagg tacgggtgca ctgcccaacg gcaccgccat aactgccgcc cccgcaacag 5821 acgacaaacc gagttctcca gtcagtgaca aacttcacgt cagggtcccc agatggtgcc 5881 cagcccatct cacccgaata agagctttcc cgcattagcg aagggctcaa gaccttgggt 5941 tcttgccgcc caccatgccc cccaccttgt ttcaacgacc tcacagcccg cctcacaagc 6001 gtcttccatt caagactcgg gaacagccgc cattttgctg cgctcccccc aacccccagt 6061 tcagggcaac cttgctcgcg gacccagact acagcccttg gcggtctctc cacacgcttc 6121 cgtcccaccg agcggcccgg cggccacgaa agccccggcc agcccagcag cccgctactc 6181 accaagtgac gatcacagcg atccacaaac aagaaccgcg acccaaatcc cggctgcgac 6241 ggaactagct gtgccacacc cggcgcgtcc ttatataatc atcggcgttc accgccccac 6301 ggagatccct ccgcagaatc gccgagaagg gactactttt cctcgcctgt tccgctctct 6361 ggaaagaaaa ccagtgccct agagtcaccc aagtcccgtc ctaaaatgtc cttctgctga 6421 tactggggtt ctaaggccga gtcttatgag cagcgggccg ctgtcctgag cgtccgggcg 6481 gaaggatcag gacgctcctg cgcccttcgt ctgacgtggc agcgctcgcc gtgacgaggg 6541 gggcgcccgc gggaggcgcc aaaacccggc gcggaggcca ggcgcgcccc gatttcgata 6601 ttcctcggat ctcggataga tcgggattga ggatgtaggt tgaacttctc tcgggtgatc 6661 ccgatctcgg atattcggga ttggagtaat acctggcgat aggcgctgcc aagcaatagc 6721 caaggcgcaa ggccggccaa gagacttgcg atagttcgtt cgagaggtag cctgcaagat 6781 ggatcggacg aaaggtttga agattttccg acgagggcct tggagagttc ggaaatttaa 6841 atgcaactat ggatgaac //