LOCUS       sequence-382448-        2159 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 nsp10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2159)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2159)
  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-m4046840
FEATURES             Location/Qualifiers
     source          1..2159
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             509..529
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             530..553
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     rep_origin      674..1262
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1163..1182
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1368..2027
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1434..1453)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2098..2132
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag actcttctat ggccggtaac gcaacggaag ttcccgcgaa
      121 tagcacagtg ctgtcttttt gcgctttcgc tgtagacgcg gcaaaagctt acaaggatta
      181 tttggcctca ggcggacagc caattaccaa ctgtgtcaaa atgctctgca ctcacaccgg
      241 gactggccaa gctataaccg taactcctga ggcaaacatg gaccaggaat ccttcggtgg
      301 tgcgagttgc tgcctgtact gtcgctgcca tatcgatcac ccgaacccga aaggcttttg
      361 tgacctgaag gggaaatacg ttcagatccc gactacgtgc gctaacgatc cggttggttt
      421 caccctgaaa aacaccgtgt gcaccgtgtg cggtatgtgg aaaggctatg gctgttcttg
      481 tgaccagctg agggaaccga tgcttcaaga gaacctgtac tttcaaagct ggagccatcc
      541 gcagtttgaa aaataaccgg cttatcggtc agtttcacct gatttacgta aaaacccgct
      601 tcggcgggtt tttgcttttg gaggggcaga aagatgaatg actgtccacg acgctatacc
      661 caaaagaaaa agcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
      721 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      781 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
      841 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      901 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      961 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1021 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1081 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1141 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1201 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     1261 aattgatcgg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg
     1321 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
     1381 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
     1441 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
     1501 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     1561 gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag ctggtgatat
     1621 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
     1681 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
     1741 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
     1801 tttcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
     1861 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
     1921 tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc
     1981 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt gatatcgagc
     2041 tcgcttggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
     2101 agaacgctcg gttgccgccg ggcgtttttt attggtcatc gatctcgatc ccgcgaaat
//
