LOCUS Exported 4764 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 leader protein. ACCESSION . VERSION . KEYWORDS pGBW-m4134160 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4764) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4764) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4764 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 542..562 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 563..580 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 633..754 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(644..663) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 698..717 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 869..915 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 945..1081 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 1126..1197 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1423..2011) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1503..1522) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2757..3617 /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(2975..2994) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 3630..3766 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 3918..3989 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 3994..4040 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 4058..4106 /note="synthetic polyadenylation signal" enhancer 4187..4490 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4492..4694 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4645..4665 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 4763..8 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" ORIGIN 1 caccatggag agccttgtcc ctggtttcaa cgagaaaaca cacgtccaac tcagtttgcc 61 tgttttacag gttcgcgacg tgctcgtacg tggctttgga gactccgtgg aggaggtctt 121 atcagaggca cgtcaacatc ttaaagatgg cacttgtggc ttagtagaag ttgaaaaagg 181 cgttttgcct caacttgaac agccctatgt gttcatcaaa cgttcggatg ctcgaactgc 241 acctcatggt catgttatgg ttgagctggt agcagaactc gaaggcattc agtacggtcg 301 tagtggtgag acacttggtg tccttgtccc tcatgtgggc gaaataccag tggcttaccg 361 caaggttctt cttcgtaaga acggtaataa aggagctggt ggccatagtt acggcgccga 421 tctaaagtca tttgacttag gcgacgagct tggcactgat ccttatgaag attttcaaga 481 aaactggaac actaaacata gcagtggtgt tacccgtgaa ctcatgcgtg agcttaacgg 541 agaaaacctt tactttcagt cgcaccatca ccatcaccat taagctagct tgactgactg 601 agatacagcg taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac 661 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt 721 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat 781 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg 841 tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc 901 ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa 961 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga 1021 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta 1081 aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt 1141 cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc 1201 tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag 1261 atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc 1321 ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca 1381 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 1441 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 1501 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 1561 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 1621 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 1681 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 1741 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 1801 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 1861 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 1921 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 1981 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 2041 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 2101 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 2161 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 2221 ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc 2281 ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc 2341 gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta 2401 ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct 2461 caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggtgtaccga 2521 gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc 2581 ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac 2641 ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc 2701 gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga 2761 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt 2821 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag 2881 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag 2941 aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta 3001 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg 3061 agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca 3121 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgattggag 3181 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc 3241 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg 3301 tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc 3361 ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg 3421 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg 3481 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga 3541 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac 3601 tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga 3661 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac 3721 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt 3781 taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg 3841 cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca 3901 gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt 3961 ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga 4021 ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc 4081 catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt 4141 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta 4201 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga 4261 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt 4321 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta 4381 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg 4441 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt 4501 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc 4561 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat 4621 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct 4681 atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca 4741 ctcgacacac ccgccagcgg ccgc //