LOCUS Exported 6840 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp7. ACCESSION . VERSION . KEYWORDS pGBW-m4134097 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6840) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 6840) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6840 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 81..384 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 386..588 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 539..559 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 915..935 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 936..962 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 1031..1152 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1042..1061) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1096..1115 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 1267..1313 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 1343..1479 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 1524..1595 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1821..2409) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1901..1920) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 2909..4118 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 3955..3974 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 4122..5150 /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" polyA_signal 5177..5225 /note="synthetic polyadenylation signal" CDS 5491..6351 /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(5709..5728) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 6364..6500 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 6652..6723 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 6728..6774 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 6792..6840 /note="synthetic polyadenylation signal" ORIGIN 1 atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat 61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg 121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata 181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta 241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc 301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 361 gtattagtca tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga 421 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg 481 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg 541 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac 601 cgtcagatca gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgcca 661 ccatgagcaa aatgtctgac gtcaagtgta catccgtagt tctgttatca gtgcttcaac 721 agctgcgggt ggagagttcg tcaaagctct gggcccagtg cgtgcagctg cacaatgata 781 tattgctcgc gaaagatact acggaagctt tcgagaagat ggtcagcttg ctatctgttc 841 tactgagcat gcaaggagca gtagacatca acaaactttg cgaagagatg ttagacaacc 901 gagccaccct gcaagaaaac ctttactttc agtcgtaccc ctatgacgta ccagattatg 961 cgtaaggatc cgcaggcctc tgctagcttg actgactgag atacagcgta ccttcagctc 1021 acagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa 1081 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc 1141 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg 1201 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg 1261 gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct 1321 cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg 1381 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca 1441 cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg 1501 cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg 1561 tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc 1621 gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac 1681 tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg 1741 attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 1801 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 1861 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 1921 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 1981 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 2041 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 2101 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 2161 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 2221 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc 2281 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 2341 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 2401 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 2461 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 2521 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 2581 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 2641 gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt 2701 ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg 2761 cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg 2821 agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc 2881 cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg 2941 ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg 3001 atccttccgc ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac 3061 cccagtatca gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt 3121 tctttccaga gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg 3181 gatctccgtg gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc 3241 tagttccgtc gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc 3301 acttggtgag tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg 3361 tgggacggaa gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt 3421 gccctgaact gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat 3481 ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc 3541 ggcaagaacc caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga 3601 tgggctgggc accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt 3661 ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct 3721 ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag 3781 ggtggggcca cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt 3841 cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat 3901 aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc 3961 tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac 4021 cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt 4081 ccgctaaatt ctggccgttt ttggcttttt tgttagacaa catgggtaaa aagcctgaac 4141 tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga 4201 tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat 4261 atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc 4321 actttgcatc cgccgcgctc ccgattccgg aagtgcttga cattggggag ttcagcgaga 4381 gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa 4441 ccgaactgcc cgctgttctg cagccggtcg cggaggcaat ggatgccatc gctgccgccg 4501 atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta 4561 catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga 4621 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg 4681 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga 4741 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc 4801 aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga 4861 cgcgctactt cgagcggagg catccggagc ttgcaggatc gccccggctc cgggcgtata 4921 tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg 4981 cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc 5041 gtacacaaat cgcccgcaga agcgccgccg tctggaccga tggctgtgta gaagtactcg 5101 ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaataa gctagtatgt 5161 aagcctagtc ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt 5221 gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg 5281 acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt 5341 ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct 5401 agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat 5461 caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt 5521 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa 5581 gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac 5641 agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt 5701 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt 5761 cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat 5821 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac 5881 actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg 5941 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc 6001 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa 6061 ctattaactg gcgaactact tactctagct tcccggcaac agttgataga ctggatggag 6121 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct 6181 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat 6241 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa 6301 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta accgattcta 6361 ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata 6421 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct 6481 taccagaaat ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat 6541 ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa 6601 tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga 6661 ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct 6721 ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc 6781 gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt //