LOCUS       sequence-287802-        2625 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 2625)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2625)
  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-m4046827
FEATURES             Location/Qualifiers
     source          1..2625
                     /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             999..1019
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1020..1046
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      1163..1751
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1652..1671
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1857..2516
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1923..1942)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2587..2621
                     /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 agtcaagcgt
      121 ggcaaccggg tgttgctatg cctaatcttt acaaaatgca aagaatgcta ttagaaaagt
      181 gtgaccttca aaattatggt gatagtgcaa cattacctaa aggcataatg atgaatgtcg
      241 caaaatatac tcaactgtgt caatatttaa acacattaac attagctgta ccctataata
      301 tgagagttat acattttggt gctggttctg ataaaggagt tgcaccaggt acagctgttt
      361 taagacagtg gttgcctacg ggtacgctgc ttgtcgattc agatcttaat gactttgtct
      421 ctgatgcaga ttcaactttg attggtgatt gtgcaactgt acatacagct aataaatggg
      481 atctcattat tagtgatatg tacgacccta agactaaaaa tgttacaaaa gaaaatgact
      541 ctaaagaggg ttttttcact tacatttgtg ggtttataca acaaaagcta gctcttggag
      601 gttccgtggc tataaagata acagaacatt cttggaatgc tgatctttat aagctcatgg
      661 gacacttcgc atggtggaca gcctttgtta ctaatgtgaa tgcgtcatca tctgaagcat
      721 ttttaattgg atgtaattat cttggcaaac cacgcgaaca aatagatggt tatgtcatgc
      781 atgcaaatta catattttgg aggaatacaa atccaattca gttgtcttcc tattctttat
      841 ttgacatgag taaatttccc cttaaattaa ggggtactgc tgttatgtct ttaaaagaag
      901 gtcaaatcaa tgatatgatt ttatctcttc ttagtaaagg tagacttata attagagaaa
      961 acaacagagt tgttatttct agtgatgttc ttgttaacga gaacctgtac tttcaaagct
     1021 acccgtatga cgttcctgat tatgcctaac cggcttatcg gtcagtttca cctgatttac
     1081 gtaaaaaccc gcttcggcgg gtttttgctt ttggaggggc agaaagatga atgactgtcc
     1141 acgacgctat acccaaaaga aattgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     1321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1741 gcctatggaa attgatcggg cacgtaagag gttccaactt tcaccataat gaaataagat
     1801 cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg
     1861 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
     1921 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta
     1981 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
     2041 ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc
     2101 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt
     2161 tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc
     2221 aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata
     2281 tgtttttcgt ttcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca
     2341 atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca
     2401 aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg
     2461 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttg
     2521 atatcgagct cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg
     2581 gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggt
//
