LOCUS       sequence-300256-        2330 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 2330)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2330)
  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-m4046963
FEATURES             Location/Qualifiers
     source          1..2330
                     /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..743
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      864..1452
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1353..1372
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1558..2217
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1624..1643)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2288..2322
                     /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 tttcaaagcg
      721 actacaagga tgatgatgac aaataaccgg cttatcggtc agtttcacct gatttacgta
      781 aaaacccgct tcggcgggtt tttgcttttg gaggggcaga aagatgaatg actgtccacg
      841 acgctatacc caaaagaaaa agcttgagat cctttttttc tgcgcgtaat ctgctgcttg
      901 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
      961 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     1021 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1081 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     1141 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     1201 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     1261 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     1321 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     1381 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     1441 agcctatgga aattgatcgg gcacgtaaga ggttccaact ttcaccataa tgaaataaga
     1501 tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg
     1561 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat
     1621 tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt
     1681 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
     1741 attcttgccc gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag
     1801 ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg
     1861 ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca catatattcg
     1921 caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat
     1981 atgtttttcg tttcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc
     2041 aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac
     2101 aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc
     2161 ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt
     2221 gatatcgagc tcgcttggac tcctgttgat agatccagta atgacctcag aactccatct
     2281 ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtcatc
//
