LOCUS       sequence-298362-        2336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 nsp8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2336)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2336)
  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-m4046804
FEATURES             Location/Qualifiers
     source          1..2336
                     /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             705..725
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             726..749
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     rep_origin      870..1458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1359..1378
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1564..2223
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1630..1649)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2294..2328
                     /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 gccattgcga
      121 gcgaattttc aagtcttcca tcctatgctg cattcgctac tgcacaagag gcttacgaac
      181 aggcggtggc caacggcgac tcggaagtcg ttctgaagaa actgaaaaaa tctttaaatg
      241 ttgcgaaaag cgaatttgat cgtgacgctg cgatgcagcg caagttggag aaaatggctg
      301 atcaggcaat gacgcaaatg tacaaacagg ctcgttccga agacaaaagg gccaaagtaa
      361 cctctgctat gcagaccatg ctgttcacaa tgctgcgcaa gctggataac gacgcgctca
      421 acaacatcat caacaatgca cgtgatggtt gcgtacctct caacattatc ccgcttacta
      481 ctgctgcaaa actgatggtg gttataccgg actacaacac ctataagaac acctgtgatg
      541 ggacgacttt cacctacgcc agcgcactgt gggaaatcca gcaggttgtc gatgctgaca
      601 gtaaaatcgt tcagctgagc gagatttcta tggacaattc tccgaacctg gcgtggccac
      661 tgatcgtaac tgctctgcgt gcgaatagcg cggtcaaatt acaagagaac ctgtactttc
      721 aaagctggag ccatccgcag tttgaaaaat aaccggctta tcggtcagtt tcacctgatt
      781 tacgtaaaaa cccgcttcgg cgggtttttg cttttggagg ggcagaaaga tgaatgactg
      841 tccacgacgc tatacccaaa agaaaaagct tgagatcctt tttttctgcg cgtaatctgc
      901 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
      961 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     1021 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     1081 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     1141 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     1201 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     1261 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     1321 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     1381 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     1441 gggcggagcc tatggaaatt gatcgggcac gtaagaggtt ccaactttca ccataatgaa
     1501 ataagatcac taccgggcgt attttttgag ttatcgagat tttcaggagc taaggaagct
     1561 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa
     1621 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg
     1681 gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt
     1741 attcacattc ttgcccgcct gatgaatgct catccggaat ttcgtatggc aatgaaagac
     1801 ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact
     1861 gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata
     1921 tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt
     1981 gagaatatgt ttttcgtttc agccaatccc tgggtgagtt tcaccagttt tgatttaaac
     2041 gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa
     2101 ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc
     2161 catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg
     2221 taatttgata tcgagctcgc ttggactcct gttgatagat ccagtaatga cctcagaact
     2281 ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtcatc
//
