LOCUS       sequence-298327-        2405 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 2405)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2405)
  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-m4046904
FEATURES             Location/Qualifiers
     source          1..2405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    39..63
                     /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             78..100
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             774..794
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             795..818
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      939..1527
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1428..1447
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1633..2292
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1699..1718)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2363..2397
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
       61 tcccctctag aaataatttt gtttaacttt aagaaggaga ctcttctatg gccgattcaa
      121 acggtacgat tactgtagaa gagcttaaaa agctgttaga acaatggaat ttggtgatcg
      181 ggtttctgtt cctgacctgg atctgtctcc tgcaattcgc ttacgcgaac cgcaaccggt
      241 ttctgtatat aatcaaatta attttcctgt ggctgctgtg gcccgtcacc ctggcatgct
      301 tcgttctggc tgctgtttac agaatcaatt ggattactgg cggtatcgca atcgcgatgg
      361 catgcctagt aggcctgatg tggctgtcgt attttattgc ttccttccgt ctgtttgccc
      421 gtacccgtag catgtggtct ttcaacccag aaactaacat cctgcttaac gttcctctgc
      481 acggtacaat cctgacccgc ccgctgctgg agagcgaact ggtgattggc gctgtcatct
      541 tacgtggtca tctgcgtatt gctggccacc atttgggtcg ctgtgacatc aaagacttgc
      601 cgaaagaaat cactgttgca acctctcgta cactgtccta ctacaaactg ggggcgagtc
      661 agcgtgttgc cggcgattct ggattcgcgg cttacagccg ctatcgcatc ggtaattaca
      721 agctcaacac tgaccacagc tcctctagtg ataacattgc actgctggtg caggagaacc
      781 tgtactttca aagcgactac aaggatgatg atgacaaata accggcttat cggtcagttt
      841 cacctgattt acgtaaaaac ccgcttcggc gggtttttgc ttttggaggg gcagaaagat
      901 gaatgactgt ccacgacgct atacccaaaa gaaaaagctt gagatccttt ttttctgcgc
      961 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1021 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1081 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1141 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1201 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1261 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1321 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1381 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     1441 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     1501 tcgtcagggg ggcggagcct atggaaattg atcgggcacg taagaggttc caactttcac
     1561 cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct
     1621 aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg
     1681 catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc
     1741 gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat
     1801 ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt tcgtatggca
     1861 atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat
     1921 gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt
     1981 ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa
     2041 gggtttattg agaatatgtt tttcgtttca gccaatccct gggtgagttt caccagtttt
     2101 gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat
     2161 tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt
     2221 gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag
     2281 ggcggggcgt aatttgatat cgagctcgct tggactcctg ttgatagatc cagtaatgac
     2341 ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg
     2401 tcatc
//
