LOCUS Exported 5811 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 3'-to-5' exonuclease. ACCESSION . VERSION . KEYWORDS pGBW-m4134059 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5811) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 5811) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5811 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 1589..1609 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1610..1627 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 1680..1801 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1691..1710) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1745..1764 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 1916..1962 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 1992..2128 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 2173..2244 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(2470..3058) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2550..2569) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 3804..4664 /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(4022..4041) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 4677..4813 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 4965..5036 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 5041..5087 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 5105..5153 /note="synthetic polyadenylation signal" enhancer 5234..5537 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 5539..5741 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5692..5712 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 5810..8 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" ORIGIN 1 caccatggcc gaaaatgtca ctggcttatt taaggattgc agtaaagtga tcacagggtt 61 gcacccgacc caagcgccta cgcatctttc agtagacacc aagttcaaaa ctgagggact 121 ctgtgtggac ataccaggta ttcccaagga tatgacatac agacgcctga tctcgatgat 181 gggcttcaag atgaactatc aggttaacgg ttaccccaat atgtttatta cccgagagga 241 agctatccgg cacgtgcgtg cctggatagg gtttgacgtc gagggctgtc atgcaactag 301 ggaagctgta ggaacaaatc tgccactgca actcggcttc tctaccggag tcaacctagt 361 tgccgtgcct accgggtatg tggatacacc aaacaatact gattttagca gagtctccgc 421 aaaaccccct cccggggacc agttcaaaca cttgatcccc ctgatgtaca agggtttacc 481 atggaacgtg gtgcggatta aaatagttca gatgctcagc gacacactga agaatttaag 541 tgatagagtt gtgtttgtac tttgggccca tggcttcgag ctgacctcta tgaagtattt 601 cgttaaaatt ggacccgaac gaacgtgctg cctgtgtgac cggagggcga cgtgtttctc 661 aactgcttcc gatacctatg cctgctggca ccatagcatc ggctttgatt acgtgtacaa 721 cccttttatg attgacgtgc agcagtgggg attcactggg aacctacagt cgaatcacga 781 tctttactgt caggtccatg ggaacgcaca cgttgctagc tgcgacgcaa tcatgacacg 841 ttgccttgct gtccacgagt gttttgtgaa acgagtggac tggactattg agtatcctat 901 cattggcgac gaactgaaga taaatgcggc ttgtaggaag gtgcagcata tggtcgtcaa 961 agccgccctc ttagctgata agttcccggt cctgcacgat attggcaatc caaaggccat 1021 caaatgcgta ccccaggcag atgttgagtg gaagttttac gacgcgcaac catgctccga 1081 taaggcctat aaaatcgaag agctcttcta cagttatgcc acccattctg acaagtttac 1141 cgacggtgtg tgtttgtttt ggaattgcaa cgtggatcgc taccctgcaa actctatcgt 1201 ttgtcgcttc gacacacgcg tattgtccaa tctgaatcta cccggatgcg acggaggttc 1261 actgtatgtg aacaagcacg ccttccacac ccccgcattc gataaatccg cctttgtcaa 1321 ccttaaacaa ttacctttct tttactacag cgactctcct tgcgaatccc atggaaagca 1381 agtagtgagt gatatcgatt acgtcccact caaaagtgca acatgtatta cgagatgcaa 1441 tctcggtggt gcagtctgta gacaccatgc caatgaatat cggctgtatt tggacgccta 1501 taacatgatg atttcagctg gcttctctct ttgggtttac aagcagtttg acacgtataa 1561 tctgtggaac acctttacaa ggctgcaaga aaacctttac tttcagtcgc accatcacca 1621 tcaccattaa gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat 1681 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat 1741 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt 1801 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt 1861 ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca 1921 taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt 1981 gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact 2041 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt 2101 gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga 2161 tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt 2221 tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt 2281 aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa 2341 ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt 2401 catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 2461 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 2521 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 2581 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 2641 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 2701 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 2761 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 2821 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 2881 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa 2941 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 3001 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 3061 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 3121 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 3181 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 3241 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa 3301 atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct 3361 tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta 3421 ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc 3481 acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc 3541 gaaatcgggg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc 3601 atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg 3661 tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt 3721 gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga 3781 caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct 3841 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag 3901 atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta 3961 agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc 4021 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca 4081 tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg 4141 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg 4201 ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca 4261 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa 4321 acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa 4381 ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg gaggcggata 4441 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat 4501 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc 4561 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata 4621 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat 4681 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga 4741 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga 4801 aatttatcct taagatcccg aatcgtttaa actcgactct ggctctatcg aatctccgtc 4861 gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc 4921 agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag 4981 ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt 5041 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt 5101 tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac 5161 tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat 5221 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 5281 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 5341 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 5401 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 5461 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 5521 tcatcgctat taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 5581 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 5641 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 5701 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga 5761 tcagatcttt gtcgatccta ccatccactc gacacacccg ccagcggccg c //