LOCUS Exported 6852 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp7. ACCESSION . VERSION . KEYWORDS pGBW-m4252917 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6852) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6852) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6852 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 29..77 /note="synthetic polyadenylation signal" terminator 95..141 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" misc_feature 146..217 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 369..505 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" CDS complement(518..1378) /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 1141..1160 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" polyA_signal 1644..1692 /note="synthetic polyadenylation signal" CDS complement(1719..2747) /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(2751..3960) /label=UbC promoter /note="human ubiquitin C promoter" primer_bind complement(2895..2914) /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" rep_origin 4460..5048 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4949..4968 /label=pBR322ori-F /note="pBR322 origin, forward primer" misc_feature 5274..5345 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" terminator 5390..5526 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" terminator 5556..5602 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 5717..5838 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5754..5773) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5808..5827 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" CDS complement(5907..5951) /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS complement(5952..5972) /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" promoter complement(6293..6495) /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind complement(6322..6342) /label=CMV-F /note="Human CMV immediate early promoter, forward primer" enhancer 6497..6800 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 tctggctaca ctcgatcagc agttagataa taaaatcgct atccatcgaa gatggatgtg 61 tgttggtttt ttgtgtgtgt aacgcaacga ttgatagcat aaccccttgg ggcctctaaa 121 cgggtcttga ggggtttttt gaattgagag gtagctgaag ctgctgttat gaccactaga 181 gggaagaaga tacctgtgga gctaatggtc caagatgtcg ggagccgcga tcgctgccaa 241 aaaggtaagc tgacgatagc tgacgagatt cgatgcccga cgagcaaatg agcgccacgg 301 ctgttcgcgt aagctcgaaa cgacggagat tcgatagagc cagagtcgag tttaaacgat 361 tcgggatctt aaggataaat ttctggtaag gaggacacgt atggaagtgg gcaagttggg 421 gaagccgtat ccgttgctga atctggcata tgtgggagta taagacgcgc agcgtcgcat 481 caggcatttt tttctgcgcc aatgcaccta gaatcggtta ccaatgctta atcagtgagg 541 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 601 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 661 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 721 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tatcaactgt tgccgggaag 781 ctagagtaag tagttcgcca gttaatagtt tacgcaaggt tgttgccatt gctacaggca 841 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 901 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccaa 961 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 1021 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgaa tactcaacca 1081 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 1141 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaag cgttcttcgg 1201 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgcg 1261 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 1321 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatgc 1381 gaaagcctcc tgctgatcct gtctcttgat agatgtttgc aaaagctagg actcgattat 1441 acgttgaccg atcagactat caatatctag gtttaaacag atcggtacgc tgccgtgacc 1501 aggcttgagt acaatatctg acgattggac ttcctgggtc ccaagctgga ttccgactgg 1561 ctgactgcca agcaacgata tggatcgtcg agactcgcac tgagaggatc gttctcggta 1621 caccaggcgc gcctagcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat 1681 agcgatttta ttatctaaga ctaggcttac atactagctt attcctttgc cctcggacga 1741 gtgctggggc gtcggtttcc actatcggcg agtacttcta cacagccatc ggtccagacg 1801 gcggcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt 1861 gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac caagctctga 1921 tagagttggt caagaccaat gcggagcata tacgcccgga gccggggcga tcctgcaagc 1981 tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc 2041 cagaagaaga tgttggcgac ctcgtattgg gaatccccga acatcgcctc gctccagtca 2101 atgaccgctg ttatgcggcc attgtccgtc aggacattgt tggagccgaa atccgcgtgc 2161 acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca tcagctcatc gagagcctgc 2221 gcgacggacg cactgacggt gtcgtccatc acagtttgcc agtgatacac atggggatca 2281 gcaatcgcgc atatgaaatc acgccatgta gtgtattgac cgattccttg cggtccgaat 2341 gggccgaacc cgctcgtctg gctaagatcg gcggcagcga tggcatccat tgcctccgcg 2401 accggctgca gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt 2461 gcacggcggg agatgcaata ggtcaggctc tcgctgaact ccccaatgtc aagcacttcc 2521 ggaatcggga gcgcggcgga tgcaaagtgc cgataaacat aacgatcttt gtagaaacca 2581 tcggcgcagc tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca 2641 cgagattctt cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc 2701 agaaacttct cgacagacgt cgcggtgagt tcaggctttt tacccatgtt gtctaacaaa 2761 aaagccaaaa acggccagaa tttagcggac aatttacaag tctaacactg aaaattacat 2821 attgacccaa atgattacat ttcaaaaggt gcctaaaaaa cttcacaaaa cacactcgcc 2881 aaccccgagc gcatagttca aaaccggagc ttcagctact taagaagata ggtacataaa 2941 accgaccaaa gaaactgacg cctcacttat ccctcccctc accagaggtc cggcgcctgt 3001 cgattcagga gagcctaccc taggcccgaa ccctgcgtcc tgcgacggag aaaagcctac 3061 cgcacaccta ccggctggtg gccccaccct gcattataag ccaacagaac gggtgacgtc 3121 acgacacgac gagggcgcgc gctcccaaag gtacgggtgc actgcccaac ggcaccgcca 3181 taactgccgc ccccgcaaca gacgacaaac cgagttctcc agtcagtgac aaacttcacg 3241 tcagggtccc cagatggtgc ccagcccatc tcacccgaat aagagctttc ccgcattagc 3301 gaagggctca agaccttggg ttcttgccgc ccaccatgcc ccccaccttg tttcaacgac 3361 ctcacagccc gcctcacaag cgtcttccat tcaagactcg ggaacagccg ccattttgct 3421 gcgctccccc caacccccag ttcagggcaa ccttgctcgc ggacccagac tacagccctt 3481 ggcggtctct ccacacgctt ccgtcccacc gagcggcccg gcggccacga aagccccggc 3541 cagcccagca gcccgctact caccaagtga cgatcacagc gatccacaaa caagaaccgc 3601 gacccaaatc ccggctgcga cggaactagc tgtgccacac ccggcgcgtc cttatataat 3661 catcggcgtt caccgcccca cggagatccc tccgcagaat cgccgagaag ggactacttt 3721 tcctcgcctg ttccgctctc tggaaagaaa accagtgccc tagagtcacc caagtcccgt 3781 cctaaaatgt ccttctgctg atactggggt tctaaggccg agtcttatga gcagcgggcc 3841 gctgtcctga gcgtccgggc ggaaggatca ggacgctcct gcgcccttcg tctgacgtgg 3901 cagcgctcgc cgtgacgagg ggggcgcccg cgggaggcgc caaaacccgg cgcggaggcc 3961 aggcgcgccc cgatttcgat attcctcgga tctcggatag atcgggattg aggatgtagg 4021 ttgaacttct ctcgggtgat cccgatctcg gatattcggg attggagtaa tacctggcga 4081 taggcgctgc caagcaatag ccaaggcgca aggccggcca agagacttgc gatagttcgt 4141 tcgagaggta gcctgcaaga tggatcggac gaaaggtttg aagattttcc gacgagggcc 4201 ttggagagtt cggaaattta aatgcaacta tggatgaacg aaatagacag atcgctgaga 4261 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt 4321 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata 4381 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag 4441 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 4501 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 4561 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc 4621 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 4681 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 4741 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 4801 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 4861 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 4921 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 4981 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 5041 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg 5101 ctcacatgat ttaaatgaaa ctcgtaatcg gagaacgggt gcaatagcaa ggaccagatc 5161 gaacttgact gacgcaattc tgagcgtagt cgaaacccag acaagtcgat ctcgccaagt 5221 agctgacttg actggttaat taagtttcga tagcccaagg taaccaatga gattagggag 5281 ggaagaaagt ttgatgaact acttctgacc cctagtggtg tccagaaaag accattaaag 5341 gaatgtcttg atcgaaatcg agagatcgcc tgcaggatag ctgacgacct taaggataaa 5401 tttctggtaa ggaggacacg tatggaagtg ggcaagttgg ggaagccgta tccgttgctg 5461 aatctggcat atgtgggagt ataagacgcg cagcgtcgca tcaggcattt ttttctgcgc 5521 caatgccggt agatagcaat ccggcccgag gggcacaaaa aacccctcaa gacccgttta 5581 gaggccccaa ggggttatgc tacgataccg atagagatgg gccaatacca catttgtaga 5641 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa 5701 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag 5761 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 5821 actcatcaat gtatcttatc atgtctgtga gctgaaggta cgctgtatct cagtcagtca 5881 agctagcaga ggcctgcgga tccttaactg tccatgtgct gcctctcgaa cttagctgcc 5941 gcggtttctt tcgactgaaa gtaaaggttt tctaaggttg ccctgttgtc cagcatttct 6001 tcacaaagct tgtttatgtc tacagcaccc tgcatggaaa gcaaaacaga aagtagtgaa 6061 accatttttt caaaggcttc agtagtatct ttagctaaga gaatgtcatt gtgtaactgg 6121 acacattgag cccacaattt agatgatgat tctactctga gttgttgcaa aactgagagt 6181 aagactactg atgtgcactt tacatctgac attttcatgg tggcggccgc tggcgggtgt 6241 gtcgagtgga tggtaggatc gacaaagatc tgatctgacg gttcactaaa ccagctctgc 6301 ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt 6361 tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa 6421 tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact 6481 gccaaaaccg catcagcatg gtaatagcga tgactaatac gtagatgtac tgccaagtag 6541 gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga 6601 cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt 6661 accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa 6721 catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat 6781 ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa 6841 ttgattacta ga //