LOCUS Exported 7352 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 N (Nucleocapsid). ACCESSION . VERSION . KEYWORDS pGBW-m4134905 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7352) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 7352) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7352 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 1314..1435 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1325..1344) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1379..1398 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 1550..1596 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 1626..1762 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 1807..1878 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(2104..2692) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2184..2203) /label=pBR322ori-F /note="pBR322 origin, forward primer" promoter 3192..4401 /label=UbC promoter /note="human ubiquitin C promoter" primer_bind 4238..4257 /label=hUBCpro-F /note="Human Ubiquitin C (UbC) promoter, forward primer" CDS 4405..5004 /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(4405..4424) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4901..4921 /label=Puro-F /note="Puromycin resistance gene, forward primer" polyA_signal 5031..5079 /note="synthetic polyadenylation signal" CDS 5345..6205 /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(5563..5582) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 6218..6354 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 6506..6577 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 6582..6628 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 6646..6694 /note="synthetic polyadenylation signal" enhancer 6775..7078 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 7080..7282 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 7233..7253 /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 gcttttggca 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 ctaagctagc ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata 1321 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga 1381 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa 1441 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag 1501 caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc 1561 cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc 1621 taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca 1681 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc 1741 cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga 1801 tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca 1861 aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag 1921 tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt 1981 cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta 2041 aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 2101 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 2161 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 2221 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 2281 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 2341 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 2401 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 2461 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 2521 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 2581 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 2641 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 2701 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 2761 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 2821 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 2881 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc 2941 gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg 3001 ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt 3061 ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga 3121 gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc 3181 ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc ccctcgtcac 3241 ggcgagcgct gccacgtcag acgaagggcg caggagcgtc ctgatccttc cgcccggacg 3301 ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag 3361 gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga 3421 acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt 3481 gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg 3541 ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt gagtagcggg 3601 ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg 3661 gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga actgggggtt 3721 ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac gcttgtgagg 3781 cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga acccaaggtc 3841 ttgagccctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg ggcaccatct 3901 ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg 3961 ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag cgcgcgccct 4021 cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg ccacctgccg 4081 gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct agggtaggct 4141 ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag gcgtcagttt 4201 ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt ttgaactatg 4261 cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga aatgtaatca 4321 tttgggtcaa tatgtaattt tcagtgttag acttgtaaat tgtccgctaa attctggccg 4381 tttttggctt ttttgttaga caacatgacc gagtacaagc ccacggtgcg cctcgccacc 4441 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 4501 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 4561 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg 4621 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 4681 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggagggcctc 4741 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 4801 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggccgccgag 4861 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 4921 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 4981 atgacccgca agcccggtgc ctaggctagt atgtaagcct agtcttagat aataaaatcg 5041 ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg 5101 tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg cttggcagtc 5161 agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt gtactcaagc 5221 ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg atcggtcaac 5281 gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag caggaggctt 5341 tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct 5401 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg 5461 tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg 5521 ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt 5581 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga 5641 cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga 5701 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac 5761 gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg 5821 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac 5881 gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac tacttactct 5941 agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag gaccacttct 6001 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg 6061 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat 6121 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg 6181 tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc 6241 tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct 6301 tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaagatccc 6361 gaatcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac 6421 agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct 6481 ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc 6541 cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc 6601 cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa 6661 cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca 6721 gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac 6781 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc 6841 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt 6901 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac 6961 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac 7021 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct 7081 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc 7141 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt 7201 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg 7261 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct 7321 accatccact cgacacaccc gccagcggcc gc //