LOCUS       sequence-352066-        2408 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 2408)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2408)
  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-m4046926
FEATURES             Location/Qualifiers
     source          1..2408
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             755..775
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             776..802
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      923..1511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1412..1431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1617..2276
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1683..1702)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2347..2381
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag actcttctat ggccgattca aacggtacga ttactgtaga
      121 agagcttaaa aagctgttag aacaatggaa tttggtgatc gggtttctgt tcctgacctg
      181 gatctgtctc ctgcaattcg cttacgcgaa ccgcaaccgg tttctgtata taatcaaatt
      241 aattttcctg tggctgctgt ggcccgtcac cctggcatgc ttcgttctgg ctgctgttta
      301 cagaatcaat tggattactg gcggtatcgc aatcgcgatg gcatgcctag taggcctgat
      361 gtggctgtcg tattttattg cttccttccg tctgtttgcc cgtacccgta gcatgtggtc
      421 tttcaaccca gaaactaaca tcctgcttaa cgttcctctg cacggtacaa tcctgacccg
      481 cccgctgctg gagagcgaac tggtgattgg cgctgtcatc ttacgtggtc atctgcgtat
      541 tgctggccac catttgggtc gctgtgacat caaagacttg ccgaaagaaa tcactgttgc
      601 aacctctcgt acactgtcct actacaaact gggggcgagt cagcgtgttg ccggcgattc
      661 tggattcgcg gcttacagcc gctatcgcat cggtaattac aagctcaaca ctgaccacag
      721 ctcctctagt gataacattg cactgctggt gcaggagaac ctgtactttc aaagctaccc
      781 gtatgacgtt cctgattatg cctaaccggc ttatcggtca gtttcacctg atttacgtaa
      841 aaacccgctt cggcgggttt ttgcttttgg aggggcagaa agatgaatga ctgtccacga
      901 cgctataccc aaaagaaaaa gcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
      961 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1021 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     1081 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1141 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1201 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1261 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1321 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1381 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1441 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1501 gcctatggaa attgatcggg cacgtaagag gttccaactt tcaccataat gaaataagat
     1561 cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg
     1621 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
     1681 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta
     1741 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
     1801 ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc
     1861 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt
     1921 tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc
     1981 aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata
     2041 tgtttttcgt ttcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca
     2101 atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca
     2161 aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg
     2221 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttg
     2281 atatcgagct cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg
     2341 gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtcatcg atctcgatcc
     2401 cgcgaaat
//
