LOCUS       sequence-292492-        1775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1775)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 1775)
  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-m4046971
FEATURES             Location/Qualifiers
     source          1..1775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(90..107)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(108..128)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     RBS             178..200
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    215..239
                     /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)."
     primer_bind     complement(239..258)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(240..258)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(286..320)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(391..1050)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     965..984
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(1156..1744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1236..1255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtcattcatc tttctgcccc tccaaaagca aaaacccgcc gaagcgggtt tttacgtaaa
       61 tcaggtgaaa ctgaccgata agccggttaa tgatggtgat gatggtggct ttgaaagtac
      121 aggttctcta cggcgaaacc attgagaaaa ctttgcgcat cagcgctcat agaagagtct
      181 ccttcttaaa gttaaacaaa attatttcta gaggggaatt gttatccgct cacaattccc
      241 ctatagtgag tcgtattaat ttcgcgggat cgagatcgat gaccaataaa aaacgcccgg
      301 cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc
      361 aacaggagtc caagcgagct cgatatcaaa ttacgccccg ccctgccact catcgcagta
      421 ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac
      481 ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa
      541 aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac
      601 ccagggattg gctgaaacga aaaacatatt ctcaataaac cctttaggga aataggccag
      661 gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc
      721 gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca
      781 agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac gaaattccgg
      841 atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt
      901 tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca
      961 ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac
     1021 ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga
     1081 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     1141 tacgtgcccg atcaatttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1201 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1261 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1321 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1381 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1441 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1501 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1561 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1621 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1681 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1741 tcaagctttt tcttttgggt atagcgtcgt ggaca
//
