LOCUS Exported 7400 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 N (nucleocapsid). ACCESSION . VERSION . KEYWORDS pGBW-m4134375 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7400) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 7400) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7400 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 1262..1282 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1283..1309 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 1362..1483 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1373..1392) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1427..1446 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 1598..1644 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 1674..1810 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 1855..1926 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(2152..2740) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2232..2251) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 3240..4449 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 4286..4305 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 4453..5052 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(4453..4472) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4949..4969 /label=Puro-F /note="Puromycin resistance gene, forward primer" polyA_signal 5079..5127 /note="synthetic polyadenylation signal" CDS 5393..6253 /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(5611..5630) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 6266..6402 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 6554..6625 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 6630..6676 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 6694..6742 /note="synthetic polyadenylation signal" enhancer 6823..7126 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 7128..7330 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 7281..7301 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgtct gataatggac cccaaaatca gcgaaatgca ccccgcatta cgtttggtgg 61 accctcagat tcaactggca gtaaccagaa tggagaacgc agtggggcgc gatcaaaaca 121 acgtcggccc caaggtttac ccaataatac tgcgtcttgg ttcaccgctc tcactcaaca 181 tggcaaggaa gaccttaaat tccctcgagg acaaggcgtt ccaattaaca ccaatagcag 241 tccagatgac caaattggct actaccgaag agctaccaga cgaattcgtg gtggtgacgg 301 taaaatgaaa gatctcagtc caagatggta tttctactac ctaggaactg ggccagaagc 361 tggacttccc tatggtgcta acaaagacgg catcatatgg gttgcaactg agggagcctt 421 gaatacacca aaagatcaca ttggcacccg caatcctgct aacaatgctg caatcgtgct 481 acaacttcct caaggaacaa cattgccaaa aggcttctac gcagaaggga gcagaggcgg 541 cagtcaagcc tcttctcgtt cctcatcacg tagtcgcaac agttcaagaa attcaactcc 601 aggcagcagt aggggaactt ctcctgctag aatggctggc aatggcggtg atgctgctct 661 tgctttgctg ctgcttgaca gattgaacca gcttgagagc aaaatgtctg gtaaaggcca 721 acaacaacaa ggccaaactg tcactaagaa atctgctgct gaggcttcta agaagcctcg 781 gcaaaaacgt actgccacta aagcatacaa tgtaacacaa gctttcggca gacgtggtcc 841 agaacaaacc caaggaaatt ttggggacca ggaactaatc agacaaggaa ctgattacaa 901 acattggccg caaattgcac aatttgcccc cagcgcttca gcgttcttcg gaatgtcgcg 961 cattggcatg gaagtcacac cttcgggaac gtggttgacc tacacaggtg ccatcaaatt 1021 ggatgacaaa gatccaaatt tcaaagatca agtcattttg ctgaataagc atattgacgc 1081 atacaaaaca ttcccaccaa cagagcctaa aaaggacaaa aagaagaagg ctgatgaaac 1141 tcaagcctta ccgcagagac agaagaaaca gcaaactgtg actcttcttc ctgctgcaga 1201 tttggatgat ttctccaaac aattgcaaca atccatgagc agtgctgact caactcaggc 1261 cgaaaacctt tactttcagt cgtaccccta tgacgtacca gattatgcgt aagctagctt 1321 gactgactga gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt 1381 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc 1441 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat 1501 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct 1561 ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct 1621 aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg 1681 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc 1741 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat 1801 ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc 1861 tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc 1921 tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta 1981 cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg 2041 atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc 2101 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 2161 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 2221 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 2281 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 2341 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 2401 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 2461 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 2521 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 2581 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 2641 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 2701 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 2761 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 2821 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 2881 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 2941 ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag 3001 gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa 3061 cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat 3121 cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac 3181 ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg 3241 gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc ctcgtcacgg cgagcgctgc 3301 cacgtcagac gaagggcgca ggagcgtcct gatccttccg cccggacgct caggacagcg 3361 gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga cattttagga 3421 cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa 3481 aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga 3541 ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg atttgggtcg 3601 cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct gctgggctgg 3661 ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga gagaccgcca 3721 agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg ggggagcgca 3781 gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg 3841 tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt gagcccttcg 3901 ctaatgcggg aaagctctta ttcgggtgag atgggctggg caccatctgg ggaccctgac 3961 gtgaagtttg tcactgactg gagaactcgg tttgtcgtct gttgcggggg cggcagttat 4021 ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg 4081 acgtcacccg ttctgttggc ttataatgca gggtggggcc acctgccggt aggtgtgcgg 4141 taggcttttc tccgtcgcag gacgcagggt tcgggcctag ggtaggctct cctgaatcga 4201 caggcgccgg acctctggtg aggggaggga taagtgaggc gtcagtttct ttggtcggtt 4261 ttatgtacct atcttcttaa gtagctgaag ctccggtttt gaactatgcg ctcggggttg 4321 gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt tgggtcaata 4381 tgtaattttc agtgttagac ttgtaaattg tccgctaaat tctggccgtt tttggctttt 4441 ttgttagaca acatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc 4501 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc 4561 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc 4621 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg 4681 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc 4741 gagttgagcg gttcccggct ggccgcgcag caacagatgg agggcctcct ggcgccgcac 4801 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc 4861 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg ccgccgagcg cgccggggtg 4921 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc 4981 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag 5041 cccggtgcct aggctagtat gtaagcctag tcttagataa taaaatcgct atccatcgaa 5101 gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc gcgcctggtg taccgagaac 5161 gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag ccagtcggaa 5221 tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct ggtcacggca 5281 gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt ataatcgagt 5341 cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc gcatgagtat 5401 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc 5461 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg 5521 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg 5581 ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga 5641 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta 5701 ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc 5761 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga ttggaggacc 5821 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg 5881 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc 5941 aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag cttcccggca 6001 acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct 6061 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat 6121 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg 6181 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat 6241 taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct 6301 gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc 6361 ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga atcgtttaaa 6421 ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag ccgtggcgct 6481 catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt ttggcagcga 6541 tcgcggctcc cgacatcttg gaccattagc tccacaggta tcttcttccc tctagtggtc 6601 ataacagcag cttcagctac ctctcaattc aaaaaacccc tcaagacccg tttagaggcc 6661 ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa aaaaccaaca cacatccatc 6721 ttcgatggat agcgatttta ttatctaact gctgatcgag tgtagccaga tctagtaatc 6781 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt 6841 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta 6901 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg 6961 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga 7021 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt 7081 tcctacttgg cagtacatct acgtattagt catcgctatt accatgctga tgcggttttg 7141 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc 7201 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg 7261 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat 7321 aagcagagct ggtttagtga accgtcagat cagatctttg tcgatcctac catccactcg 7381 acacacccgc cagcggccgc //