LOCUS       sequence-292514-        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-m4046806
FEATURES             Location/Qualifiers
     source          1..2399
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    43..67
                     /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             82..104
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             778..798
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             799..816
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      937..1525
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1426..1445
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1631..2290
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1697..1716)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2361..2395
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 catcgatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
       61 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagactcttc tatggcagat
      121 tccaacggta ctattaccgt tgaagagctt aaaaagctcc ttgaacaatg gaacctagta
      181 ataggtttcc tattccttac atggatttgt cttctacaat ttgcctatgc caacaggaat
      241 aggtttttgt atataattaa gttaattttc ctctggctgt tatggccagt aactttagct
      301 tgttttgtgc ttgctgctgt ttacagaata aattggatca ccggtggaat tgctatcgca
      361 atggcttgtc ttgtaggctt gatgtggctc agctacttca ttgcttcttt cagactgttt
      421 gcgcgtacgc gttccatgtg gtcattcaat ccagaaacta acattcttct caacgtgcca
      481 ctccatggca ctattctgac cagaccgctt ctagaaagtg aactcgtaat cggagctgtg
      541 atccttcgtg gacatcttcg tattgctgga caccatctag gacgctgtga catcaaggac
      601 ctgcctaaag aaatcactgt tgctacatca cgaacgcttt cttattacaa attgggagct
      661 tcgcagcgtg tagcaggtga ctcaggtttt gctgcataca gtcgctacag gattggcaac
      721 tataaattaa acacagacca ttccagtagc agtgacaata ttgctttgct tgtacaggag
      781 aacctgtact ttcaaagcca ccatcatcac catcattaac cggcttatcg gtcagtttca
      841 cctgatttac gtaaaaaccc gcttcggcgg gtttttgctt ttggaggggc agaaagatga
      901 atgactgtcc acgacgctat acccaaaaga aaaagcttga gatccttttt ttctgcgcgt
      961 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     1021 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     1081 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     1141 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     1201 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     1261 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     1321 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     1381 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     1441 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     1501 gtcagggggg cggagcctat ggaaattgat cgggcacgta agaggttcca actttcacca
     1561 taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa
     1621 ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca
     1681 tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt
     1741 tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc
     1801 ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaatttc gtatggcaat
     1861 gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga
     1921 gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct
     1981 acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg
     2041 gtttattgag aatatgtttt tcgtttcagc caatccctgg gtgagtttca ccagttttga
     2101 tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta
     2161 tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga
     2221 tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg
     2281 cggggcgtaa tttgatatcg agctcgcttg gactcctgtt gatagatcca gtaatgacct
     2341 cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggt
//
