LOCUS       sequence-298351-        2342 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 2342)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2342)
  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-m4046778
FEATURES             Location/Qualifiers
     source          1..2342
                     /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..755
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     rep_origin      876..1464
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1365..1384
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1570..2229
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1636..1655)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2300..2334
                     /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 aaagcgagca gaagcttatt tccgaagaag atctctaacc ggcttatcgg tcagtttcac
      781 ctgatttacg taaaaacccg cttcggcggg tttttgcttt tggaggggca gaaagatgaa
      841 tgactgtcca cgacgctata cccaaaagaa aaagcttgag atcctttttt tctgcgcgta
      901 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
      961 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1021 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1081 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1141 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     1201 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     1261 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     1321 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     1381 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     1441 tcaggggggc ggagcctatg gaaattgatc gggcacgtaa gaggttccaa ctttcaccat
     1501 aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag
     1561 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     1621 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     1681 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     1741 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaatttcg tatggcaatg
     1801 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     1861 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     1921 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     1981 tttattgaga atatgttttt cgtttcagcc aatccctggg tgagtttcac cagttttgat
     2041 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     2101 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     2161 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     2221 ggggcgtaat ttgatatcga gctcgcttgg actcctgttg atagatccag taatgacctc
     2281 agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtca
     2341 tc
//
