LOCUS       sequence-287799-        2628 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 2628)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2628)
  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-m4046816
FEATURES             Location/Qualifiers
     source          1..2628
                     /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             1005..1025
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1026..1049
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      1166..1754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1655..1674
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1860..2519
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1926..1945)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2590..2624
                     /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 tcaagccaag
      121 catggcagcc cggggtggcc atgccaaact tgtacaaaat gcagcgtatg ttactggaga
      181 agtgcgacct gcaaaattat ggtgattctg cgacgcttcc taaaggaatt atgatgaacg
      241 tagctaaata cactcagctg tgtcagtatc tcaacaccct gacactggct gtcccgtaca
      301 acatgcgcgt tatccacttc ggcgcaggct ccgataaagg tgttgcgccg ggcactgctg
      361 tactgcgtca gtggctgccg accggtactc tgctggttga ctctgacctc aatgattttg
      421 tgagtgacgc cgattccacc ctgatcggtg attgcgctac cgttcatact gcgaacaaat
      481 gggacctgat tatcagcgat atgtacgacc cgaagaccaa aaacgtaacc aaagaaaacg
      541 actcgaagga aggcttcttt acttatatct gtggtttcat tcagcagaaa ctggctcttg
      601 gcggttcggt ggcaatcaag attacggaac actcttggaa cgctgaccta tacaaactta
      661 tgggccattt cgcgtggtgg accgcctttg ttactaatgt caacgcgagc agctctgagg
      721 cattcctgat cggatgcaat tacctgggta aaccgcgaga acagatcgat ggctatgtta
      781 tgcacgctaa ctacatattc tggcgtaaca cgaacccgat ccaattatcc tcatattccc
      841 tgtttgacat gagcaaattc ccactaaaac tgcgcggtac agcagtgatg tctctgaaag
      901 aaggccagat taacgacatg attctgagtc tgctgtctaa agggagactg atcatccgtg
      961 agaataaccg tgtggtcatc tcatccgatg ttctggtaaa caatgagaac ctgtactttc
     1021 aaagcgacta caaggatgat gatgacaaat aaccggctta tcggtcagtt tcacctgatt
     1081 tacgtaaaaa cccgcttcgg cgggtttttg cttttggagg ggcagaaaga tgaatgactg
     1141 tccacgacgc tatacccaaa agaaattgag atcctttttt tctgcgcgta atctgctgct
     1201 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     1261 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     1321 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     1381 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     1441 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     1501 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     1561 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     1621 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     1681 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     1741 ggagcctatg gaaattgatc gggcacgtaa gaggttccaa ctttcaccat aatgaaataa
     1801 gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa
     1861 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
     1921 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata
     1981 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
     2041 acattcttgc ccgcctgatg aatgctcatc cggaatttcg tatggcaatg aaagacggtg
     2101 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa
     2161 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt
     2221 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga
     2281 atatgttttt cgtttcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg
     2341 ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg
     2401 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg
     2461 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat
     2521 ttgatatcga gctcgcttgg actcctgttg atagatccag taatgacctc agaactccat
     2581 ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggt
//
