LOCUS       sequence-292478-        2399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 M (membrane).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2399)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2399)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046733
FEATURES             Location/Qualifiers
     source          1..2399
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             386..406
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             407..424
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      545..1133
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1034..1053
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1239..1898
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1305..1324)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1969..2003
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     primer_bind     2031..2050
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2031..2049
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2050..2074
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             2089..2111
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 gtattttatt gcttccttcc gtctgtttgc ccgtacccgt agcatgtggt ctttcaaccc
       61 agaaactaac atcctgctta acgttcctct gcacggtaca atcctgaccc gcccgctgct
      121 ggagagcgaa ctggtgattg gcgctgtcat cttacgtggt catctgcgta ttgctggcca
      181 ccatttgggt cgctgtgaca tcaaagactt gccgaaagaa atcactgttg caacctctcg
      241 tacactgtcc tactacaaac tgggggcgag tcagcgtgtt gccggcgatt ctggattcgc
      301 ggcttacagc cgctatcgca tcggtaatta caagctcaac actgaccaca gctcctctag
      361 tgataacatt gcactgctgg tgcaggagaa cctgtacttt caaagccacc atcatcacca
      421 tcattaaccg gcttatcggt cagtttcacc tgatttacgt aaaaacccgc ttcggcgggt
      481 ttttgctttt ggaggggcag aaagatgaat gactgtccac gacgctatac ccaaaagaaa
      541 aagcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
      601 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
      661 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
      721 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      781 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
      841 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
      901 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      961 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1021 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1081 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaattgatcg
     1141 ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt
     1201 ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     1261 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     1321 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     1381 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     1441 atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg
     1501 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     1561 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     1621 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtttcagcca
     1681 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     1741 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     1801 cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat
     1861 tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tgatatcgag ctcgcttgga
     1921 ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc
     1981 ggttgccgcc gggcgttttt tattggtcat cgatctcgat cccgcgaaat taatacgact
     2041 cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
     2101 taagaaggag actcttctat ggccgattca aacggtacga ttactgtaga agagcttaaa
     2161 aagctgttag aacaatggaa tttggtgatc gggtttctgt tcctgacctg gatctgtctc
     2221 ctgcaattcg cttacgcgaa ccgcaaccgg tttctgtata taatcaaatt aattttcctg
     2281 tggctgctgt ggcccgtcac cctggcatgc ttcgttctgg ctgctgttta cagaatcaat
     2341 tggattactg gcggtatcgc aatcgcgatg gcatgcctag taggcctgat gtggctgtc
//
