LOCUS Exported 4773 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 leader protein. ACCESSION . VERSION . KEYWORDS pGBW-m4133856 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4773) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4773) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4773 /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..589 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" polyA_signal 642..763 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(653..672) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 707..726 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 878..924 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 954..1090 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 1135..1206 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1432..2020) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1512..1531) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2766..3626 /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(2984..3003) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 3639..3775 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 3927..3998 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 4003..4049 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 4067..4115 /note="synthetic polyadenylation signal" enhancer 4196..4499 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4501..4703 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4654..4674 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 4772..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 cgtaccccta tgacgtacca gattatgcgt aagctagctt 601 gactgactga gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt 661 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc 721 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat 781 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct 841 ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct 901 aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg 961 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc 1021 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat 1081 ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc 1141 tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc 1201 tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta 1261 cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg 1321 atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc 1381 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 1441 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 1501 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 1561 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 1621 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 1681 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 1741 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 1801 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 1861 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 1921 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 1981 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 2041 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 2101 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 2161 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 2221 ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag 2281 gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa 2341 cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat 2401 cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac 2461 ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg 2521 gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt 2581 cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag 2641 cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa 2701 cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct 2761 ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc 2821 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg 2881 gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc 2941 gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat 3001 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg 3061 acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag 3121 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa 3181 cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc 3241 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca 3301 cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc 3361 tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc 3421 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg 3481 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta 3541 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag 3601 gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc 3661 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc 3721 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc 3781 cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa 3841 cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc 3901 tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt 3961 ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac 4021 ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca 4081 acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc 4141 agatctagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta 4201 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt 4261 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg 4321 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta 4381 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga 4441 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc 4501 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc 4561 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact 4621 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt 4681 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc 4741 taccatccac tcgacacacc cgccagcggc cgc //