LOCUS       sequence-292487-        2630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 2'-O-ribose 
            methyltransferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2630)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2630)
  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-m4046764
FEATURES             Location/Qualifiers
     source          1..2630
                     /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             1003..1023
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1024..1047
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     rep_origin      1168..1756
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1657..1676
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1862..2521
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1928..1947)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2592..2626
                     /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 tatgagtcaa
      121 gcgtggcaac cgggtgttgc tatgcctaat ctttacaaaa tgcaaagaat gctattagaa
      181 aagtgtgacc ttcaaaatta tggtgatagt gcaacattac ctaaaggcat aatgatgaat
      241 gtcgcaaaat atactcaact gtgtcaatat ttaaacacat taacattagc tgtaccctat
      301 aatatgagag ttatacattt tggtgctggt tctgataaag gagttgcacc aggtacagct
      361 gttttaagac agtggttgcc tacgggtacg ctgcttgtcg attcagatct taatgacttt
      421 gtctctgatg cagattcaac tttgattggt gattgtgcaa ctgtacatac agctaataaa
      481 tgggatctca ttattagtga tatgtacgac cctaagacta aaaatgttac aaaagaaaat
      541 gactctaaag agggtttttt cacttacatt tgtgggttta tacaacaaaa gctagctctt
      601 ggaggttccg tggctataaa gataacagaa cattcttgga atgctgatct ttataagctc
      661 atgggacact tcgcatggtg gacagccttt gttactaatg tgaatgcgtc atcatctgaa
      721 gcatttttaa ttggatgtaa ttatcttggc aaaccacgcg aacaaataga tggttatgtc
      781 atgcatgcaa attacatatt ttggaggaat acaaatccaa ttcagttgtc ttcctattct
      841 ttatttgaca tgagtaaatt tccccttaaa ttaaggggta ctgctgttat gtctttaaaa
      901 gaaggtcaaa tcaatgatat gattttatct cttcttagta aaggtagact tataattaga
      961 gaaaacaaca gagttgttat ttctagtgat gttcttgtta acgagaacct gtactttcaa
     1021 agctggagcc atccgcagtt tgaaaaataa ccggcttatc ggtcagtttc acctgattta
     1081 cgtaaaaacc cgcttcggcg ggtttttgct tttggagggg cagaaagatg aatgactgtc
     1141 cacgacgcta tacccaaaag aaaaagcttg agatcctttt tttctgcgcg taatctgctg
     1201 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1261 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1321 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1381 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1441 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1501 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1561 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1621 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1681 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1741 gcggagccta tggaaattga tcgggcacgt aagaggttcc aactttcacc ataatgaaat
     1801 aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa
     1861 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
     1921 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
     1981 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
     2041 tcacattctt gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg
     2101 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
     2161 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
     2221 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
     2281 gaatatgttt ttcgtttcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     2341 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
     2401 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
     2461 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     2521 atttgatatc gagctcgctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
     2581 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
//
