LOCUS       sequence-287842-        2391 bp    DNA     circular SYN 14-APR-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 2391)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2391)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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..2391
                     /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..812
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      929..1517
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1418..1437
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1623..2282
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1689..1708)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2353..2387
                     /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 aagccaccat catcaccatc attaaccggc ttatcggtca gtttcacctg
      841 atttacgtaa aaacccgctt cggcgggttt ttgcttttgg aggggcagaa agatgaatga
      901 ctgtccacga cgctataccc aaaagaaatt gagatccttt ttttctgcgc gtaatctgct
      961 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1021 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1081 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1141 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1201 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1261 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     1321 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     1381 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     1441 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     1501 ggcggagcct atggaaattg atcgggcacg taagaggttc caactttcac cataatgaaa
     1561 taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta
     1621 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     1681 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     1741 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     1801 ttcacattct tgcccgcctg atgaatgctc atccggaatt tcgtatggca atgaaagacg
     1861 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     1921 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     1981 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     2041 agaatatgtt tttcgtttca gccaatccct gggtgagttt caccagtttt gatttaaacg
     2101 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     2161 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     2221 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     2281 aatttgatat cgagctcgct tggactcctg ttgatagatc cagtaatgac ctcagaactc
     2341 catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg t
//
