LOCUS Exported 4533 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 nsp7. ACCESSION . VERSION . KEYWORDS pGBW-m4133268 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4533) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4533) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4533 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 251..271 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 272..349 /codon_start=1 /product="calmodulin-binding peptide" /label=CBP /note="derived from skeletal muscle myosin light chain kinase; binds calmodulin with nanomolar affinity in the presence of calcium" /translation="KRRWKKNFIAVSAANRFKKISSSGAL" polyA_signal 402..523 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(413..432) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 467..486 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 638..684 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 714..850 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 895..966 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(1192..1780) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1272..1291) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2526..3386 /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(2744..2763) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 3399..3535 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 3687..3758 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 3763..3809 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 3827..3875 /note="synthetic polyadenylation signal" enhancer 3956..4259 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4261..4463 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4414..4434 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 caccatgaaa atgtcagatg taaagtgcac atcagtagtc ttactctcag ttttgcaaca 61 actcagagta gaatcatcat ctaaattgtg ggctcaatgt gtccagttac acaatgacat 121 tctcttagct aaagatacta ctgaagcctt tgaaaaaatg gtttcactac tttctgtttt 181 gctttccatg cagggtgctg tagacataaa caagctttgt gaagaaatgc tggacaacag 241 ggcaacctta gaaaaccttt actttcagtc gaagaggcga tggaaaaaga attttatcgc 301 ggtgagtgca gccaacagat tcaaaaagat atcctcgtct ggtgccctct aagctagctt 361 gactgactga gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt 421 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc 481 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat 541 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct 601 ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct 661 aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg 721 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc 781 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat 841 ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc 901 tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc 961 tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta 1021 cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg 1081 atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc 1141 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 1201 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 1261 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 1321 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 1381 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 1441 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 1501 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 1561 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 1621 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 1681 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 1741 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 1801 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 1861 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 1921 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 1981 ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag 2041 gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa 2101 cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat 2161 cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac 2221 ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg 2281 gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt 2341 cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag 2401 cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa 2461 cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct 2521 ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc 2581 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg 2641 gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc 2701 gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat 2761 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg 2821 acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag 2881 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa 2941 cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc 3001 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca 3061 cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc 3121 tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc 3181 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg 3241 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta 3301 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag 3361 gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc 3421 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc 3481 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc 3541 cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa 3601 cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc 3661 tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt 3721 ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac 3781 ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca 3841 acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc 3901 agatctagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta 3961 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt 4021 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg 4081 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta 4141 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga 4201 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc 4261 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc 4321 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact 4381 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt 4441 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc 4501 taccatccac tcgacacacc cgccagcggc cgc //