LOCUS       sequence-287782-        2325 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 2325)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2325)
  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-m4046737
FEATURES             Location/Qualifiers
     source          1..2325
                     /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             699..719
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             720..746
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      863..1451
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1352..1371
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1557..2216
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1623..1642)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2287..2321
                     /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 atagcctcag
      121 agtttagttc ccttccatca tatgcagctt ttgctactgc tcaagaagct tatgagcagg
      181 ctgttgctaa tggtgattct gaagttgttc ttaaaaagtt gaagaagtct ttgaatgtgg
      241 ctaaatctga atttgaccgt gatgcagcca tgcaacgtaa gttggaaaag atggctgatc
      301 aagctatgac ccaaatgtat aaacaggcta gatctgagga caagagggca aaagttacta
      361 gtgctatgca gacaatgctt ttcactatgc ttagaaagtt ggataatgat gcactcaaca
      421 acattatcaa caatgcaaga gatggttgtg ttcccttgaa cataatacct cttacaacag
      481 cagccaaact aatggttgtc ataccagact ataacacata taaaaatacg tgtgatggta
      541 caacatttac ttatgcatca gcattgtggg aaatccaaca ggttgtagat gcagatagta
      601 aaattgttca acttagtgaa attagtatgg acaattcacc taatttagca tggcctctta
      661 ttgtaacagc tttaagggcc aattctgctg tcaaattaga gaacctgtac tttcaaagct
      721 acccgtatga cgttcctgat tatgcctaac cggcttatcg gtcagtttca cctgatttac
      781 gtaaaaaccc gcttcggcgg gtttttgctt ttggaggggc agaaagatga atgactgtcc
      841 acgacgctat acccaaaaga aattgagatc ctttttttct gcgcgtaatc tgctgcttgc
      901 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
      961 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     1021 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1081 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1141 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1201 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1261 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1321 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1381 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1441 gcctatggaa attgatcggg cacgtaagag gttccaactt tcaccataat gaaataagat
     1501 cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg
     1561 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
     1621 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta
     1681 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
     1741 ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc
     1801 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt
     1861 tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc
     1921 aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata
     1981 tgtttttcgt ttcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca
     2041 atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca
     2101 aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg
     2161 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttg
     2221 atatcgagct cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg
     2281 gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggt
//
