LOCUS       sequence-287835-        2631 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 2631)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2631)
  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-m4046858
FEATURES             Location/Qualifiers
     source          1..2631
                     /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..1052
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      1169..1757
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1658..1677
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1863..2522
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1929..1948)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2593..2627
                     /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 aaagctaccc gtatgacgtt cctgattatg cctaaccggc ttatcggtca gtttcacctg
     1081 atttacgtaa aaacccgctt cggcgggttt ttgcttttgg aggggcagaa agatgaatga
     1141 ctgtccacga cgctataccc aaaagaaatt gagatccttt ttttctgcgc gtaatctgct
     1201 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1261 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1321 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1381 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1441 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1501 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     1561 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     1621 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     1681 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     1741 ggcggagcct atggaaattg atcgggcacg taagaggttc caactttcac cataatgaaa
     1801 taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta
     1861 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     1921 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     1981 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     2041 ttcacattct tgcccgcctg atgaatgctc atccggaatt tcgtatggca atgaaagacg
     2101 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     2161 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     2221 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     2281 agaatatgtt tttcgtttca gccaatccct gggtgagttt caccagtttt gatttaaacg
     2341 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     2401 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     2461 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     2521 aatttgatat cgagctcgct tggactcctg ttgatagatc cagtaatgac ctcagaactc
     2581 catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg t
//
