LOCUS       sequence-292519-        2069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 nsp9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2069)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2069)
  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-m4046932
FEATURES             Location/Qualifiers
     source          1..2069
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    43..67
                     /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             82..104
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             448..468
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             469..486
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      607..1195
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1096..1115
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1301..1960
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1367..1386)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2031..2065
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 catcgatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
       61 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagactcttc tatgaatgag
      121 cttagtcctg ttgcactacg acagatgtct tgtgctgccg gtactacaca aactgcttgc
      181 actgatgaca atgcgttagc ttactacaac acaacaaagg gaggtaggtt tgtacttgca
      241 ctgttatccg atttacagga tttgaaatgg gctagattcc ctaagagtga tggaactggt
      301 actatctata cagaactgga accaccttgt aggtttgtta cagacacacc taaaggtcct
      361 aaagtgaagt atttatactt tattaaagga ttaaacaacc taaatagagg tatggtactt
      421 ggtagtttag ctgccacagt acgtctagag aacctgtact ttcaaagcca ccatcatcac
      481 catcattaac cggcttatcg gtcagtttca cctgatttac gtaaaaaccc gcttcggcgg
      541 gtttttgctt ttggaggggc agaaagatga atgactgtcc acgacgctat acccaaaaga
      601 aaaagcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
      661 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      721 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
      781 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      841 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      901 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
      961 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     1021 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     1081 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     1141 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaattgat
     1201 cgggcacgta agaggttcca actttcacca taatgaaata agatcactac cgggcgtatt
     1261 ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg
     1321 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
     1381 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
     1441 cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
     1501 gaatgctcat ccggaatttc gtatggcaat gaaagacggt gagctggtga tatgggatag
     1561 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag
     1621 tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta
     1681 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtttcagc
     1741 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
     1801 cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
     1861 ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga
     1921 attacaacag tactgcgatg agtggcaggg cggggcgtaa tttgatatcg agctcgcttg
     1981 gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc
     2041 tcggttgccg ccgggcgttt tttattggt
//
