LOCUS       sequence-287790-        2622 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 2622)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2622)
  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-m4046824
FEATURES             Location/Qualifiers
     source          1..2622
                     /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..1043
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1854..2513
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1920..1939)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2584..2618
                     /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 tttcaaagcg
     1021 actacaagga tgatgatgac aaataaccgg cttatcggtc agtttcacct gatttacgta
     1081 aaaacccgct tcggcgggtt tttgcttttg gaggggcaga aagatgaatg actgtccacg
     1141 acgctatacc caaaagaaat tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaatt gatcgggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac
     1801 taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct aaaatggaga
     1861 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
     1921 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
     1981 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
     2041 ttgcccgcct gatgaatgct catccggaat ttcgtatggc aatgaaagac ggtgagctgg
     2101 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
     2161 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
     2221 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
     2281 ttttcgtttc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
     2341 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
     2401 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     2461 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taatttgata
     2521 tcgagctcgc ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat
     2581 ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gt
//
