LOCUS       sequence-297845-        2624 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 2624)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2624)
  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-m4046809
FEATURES             Location/Qualifiers
     source          1..2624
                     /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..1037
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      1158..1746
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1647..1666
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1852..2511
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1918..1937)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2582..2616
                     /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 tttcaaagcc
     1021 accatcatca ccatcattaa ccggcttatc ggtcagtttc acctgattta cgtaaaaacc
     1081 cgcttcggcg ggtttttgct tttggagggg cagaaagatg aatgactgtc cacgacgcta
     1141 tacccaaaag aaaaagcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1201 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1261 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1321 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1381 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1441 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1501 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1561 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1621 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1681 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     1741 tggaaattga tcgggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta
     1801 ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa aatggagaaa
     1861 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag
     1921 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc
     1981 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
     2041 gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg
     2101 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
     2161 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
     2221 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
     2281 ttcgtttcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     2341 gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg
     2401 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga
     2461 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta atttgatatc
     2521 gagctcgctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc atctggattt
     2581 gttcagaacg ctcggttgcc gccgggcgtt ttttattggt catc
//
