LOCUS Exported 4953 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression plasmid for SARS-CoV-2 M (membrane). ACCESSION . VERSION . KEYWORDS pGBW-m4134529 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4953) TITLE SARS-CoV-2 protein Mammalian expression constructs, tagged, for protein expression and/or purification REFERENCE 2 (bases 1 to 4953) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4953 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..304 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 306..508 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 459..479 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 577..586 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1249..1269 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1270..1347 /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 1400..1521 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1411..1430) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1465..1484 /label=SV40pA-R /note="SV40 polyA, reverse primer" terminator 1636..1682 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" terminator 1712..1848 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 1893..1964 /label=3' beta-globin insulator /note="insulator downstream of the human beta-globin locus (Farrell et al., 2002)" rep_origin complement(2190..2778) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2270..2289) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 3524..4384 /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(3742..3761) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" terminator 4397..4533 /label=rrnG terminator /note="transcription terminator from the E. coli ribosomal RNA rrnG operon (Albrechtsen et al., 1991)" misc_feature 4685..4756 /label=5' beta-globin insulator /note="insulator upstream of the human beta-globin locus (Farrell et al., 2002)" terminator 4761..4807 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" polyA_signal 4825..4873 /note="synthetic polyadenylation signal" ORIGIN 1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc 541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatggcgga cagcaacggc 601 actattacag tcgaagagct taagaaatta ctcgagcagt ggaatttggt aataggattc 661 ctgtttctaa cgtggatctg tctgctgcaa ttcgcatatg ccaaccggaa tagatttctc 721 tacatcatta agttgatctt cctgtggctg ctttggcctg tgaccttggc ttgctttgtt 781 ctggccgccg tgtaccgcat aaactggatt accggtggga tcgctatcgc aatggcgtgc 841 ctcgtgggcc ttatgtggct gtcctatttt atagcctcat tccgactctt tgctcgtact 901 aggagtatgt ggtctttcaa tcccgaaaca aatattttac taaacgtccc gctgcatggg 961 accattttaa caagaccatt gctagagagc gagctcgtga tcggcgcagt tatactgcgg 1021 ggacacctca ggatcgccgg acaccatctg ggtcgctgtg atattaaaga cctgcccaag 1081 gaaataacag tggctacctc gcgtactctt tcatactata aactgggagc cagccagaga 1141 gtggccgggg attctggctt cgcagcatat agtcgataca ggatcgggaa ctacaagctg 1201 aatacggacc actcctcctc tagtgataat attgcattgc tcgtacagga aaacctttac 1261 tttcagtcga agaggcgatg gaaaaagaat tttatcgcgg tgagtgcagc caacagattc 1321 aaaaagatat cctcgtctgg tgccctctaa gctagcttga ctgactgaga tacagcgtac 1381 cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg 1441 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt 1501 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag 1561 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca 1621 tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt 1681 tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc 1741 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca 1801 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta 1861 tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac 1921 cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct 1981 tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg 2041 ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc 2101 cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg 2161 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 2221 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 2281 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 2341 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 2401 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 2461 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 2521 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 2581 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 2641 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 2701 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 2761 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 2821 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 2881 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 2941 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 3001 tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa 3061 acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg 3121 ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc 3181 gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta 3241 tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa cgatcctctc 3301 agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg 3361 ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga 3421 tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt 3481 tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta ttcaacattt 3541 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga 3601 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg gttacatcga 3661 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gctttccaat 3721 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca 3781 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt 3841 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac 3901 catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct 3961 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga 4021 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac 4081 aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc aacagttgat 4141 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg 4201 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc 4261 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc 4321 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg 4381 gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt 4441 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca 4501 tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa actcgactct 4561 ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc 4621 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc 4681 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca 4741 gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt 4801 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga 4861 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg 4921 gtcattagtt catagcccat atatggagtt ccg //