LOCUS Exported 4500 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp7. ACCESSION . VERSION . KEYWORDS pGBW-m4133989 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4500) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4500) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4500 /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..316 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" polyA_signal 369..490 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(380..399) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 434..453 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 605..651 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 681..817 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 862..933 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1159..1747) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1239..1258) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2493..3353 /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(2711..2730) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 3366..3502 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 3654..3725 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 3730..3776 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 3794..3842 /note="synthetic polyadenylation signal" enhancer 3923..4226 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4228..4430 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4381..4401 /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 gaaagaaacc gcggcagcta agttcgagag 301 gcagcacatg gacagttaag ctagcttgac tgactgagat acagcgtacc ttcagctcac 361 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa 421 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa 481 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg 541 ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt 601 atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg 661 ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg 721 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact 781 tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg 841 atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc 901 agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga 961 aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta 1021 cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat 1081 tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 1141 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 1201 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 1261 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 1321 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 1381 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 1441 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 1501 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 1561 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 1621 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 1681 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 1741 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 1801 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 1861 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 1921 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 1981 tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc 2041 gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg 2101 ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag 2161 atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg 2221 aggaatatcg aaatcggggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct 2281 cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa 2341 gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac 2401 ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct 2461 atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc 2521 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga 2581 aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca 2641 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt 2701 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg 2761 gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc 2821 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata 2881 acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt 2941 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag 3001 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta 3061 aactattaac tggcgaacta cttactctag cttcccggca acagttgata gactggatgg 3121 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg 3181 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag 3241 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg 3301 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc 3361 taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca 3421 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc 3481 cttaccagaa atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga 3541 atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg 3601 aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg 3661 gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac 3721 ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa 3781 tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta 3841 ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc 3901 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac 3961 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa 4021 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag 4081 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc 4141 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct 4201 acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg 4261 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt 4321 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga 4381 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga 4441 accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc //