LOCUS       sequence-287865-        2154 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 2154)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2154)
  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-m4046894
FEATURES             Location/Qualifiers
     source          1..2154
                     /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             528..548
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             549..575
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      692..1280
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1181..1200
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1386..2045
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1452..1471)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2116..2150
                     /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 gccggtaacg
      121 caacggaagt tcccgcgaat agcacagtgc tgtctttttg cgctttcgct gtagacgcgg
      181 caaaagctta caaggattat ttggcctcag gcggacagcc aattaccaac tgtgtcaaaa
      241 tgctctgcac tcacaccggg actggccaag ctataaccgt aactcctgag gcaaacatgg
      301 accaggaatc cttcggtggt gcgagttgct gcctgtactg tcgctgccat atcgatcacc
      361 cgaacccgaa aggcttttgt gacctgaagg ggaaatacgt tcagatcccg actacgtgcg
      421 ctaacgatcc ggttggtttc accctgaaaa acaccgtgtg caccgtgtgc ggtatgtgga
      481 aaggctatgg ctgttcttgt gaccagctga gggaaccgat gcttcaagag aacctgtact
      541 ttcaaagcta cccgtatgac gttcctgatt atgcctaacc ggcttatcgg tcagtttcac
      601 ctgatttacg taaaaacccg cttcggcggg tttttgcttt tggaggggca gaaagatgaa
      661 tgactgtcca cgacgctata cccaaaagaa attgagatcc tttttttctg cgcgtaatct
      721 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
      781 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
      841 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
      901 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
      961 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1021 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1081 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1141 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1201 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1261 gggggcggag cctatggaaa ttgatcgggc acgtaagagg ttccaacttt caccataatg
     1321 aaataagatc actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag
     1381 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     1441 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     1501 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     1561 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag
     1621 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     1681 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     1741 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     1801 ttgagaatat gtttttcgtt tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     1861 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     1921 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     1981 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     2041 cgtaatttga tatcgagctc gcttggactc ctgttgatag atccagtaat gacctcagaa
     2101 ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat tggt
//
