LOCUS       sequence-287855-        2076 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 2076)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2076)
  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-m4046969
FEATURES             Location/Qualifiers
     source          1..2076
                     /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             450..470
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             471..497
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      614..1202
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1103..1122
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1308..1967
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1374..1393)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2038..2072
                     /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 aacaatgaat
      121 tgagtccggt agcgctccgt caaatgtcgt gcgcagctgg gacgactcag acagcctgta
      181 ctgatgacaa cgctttagcc tattacaaca ccaccaaagg tggccgcttt gttctggctc
      241 tgctttccga cctgcaagat ctgaagtggg cgagattccc aaaaagcgac ggcaccggta
      301 ctatctacac ggagctggaa ccgccttgcc gattcgtcac cgatactccg aaaggtccca
      361 aggtgaaata cctatatttt attaaaggcc tgaataacct gaaccgcggc atggttctgg
      421 gttcactggc agctaccgtg cgtctgcaag agaacctgta ctttcaaagc tacccgtatg
      481 acgttcctga ttatgcctaa ccggcttatc ggtcagtttc acctgattta cgtaaaaacc
      541 cgcttcggcg ggtttttgct tttggagggg cagaaagatg aatgactgtc cacgacgcta
      601 tacccaaaag aaattgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
      661 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      721 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
      781 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      841 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      901 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      961 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1021 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1081 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1141 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     1201 aattgatcgg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg
     1261 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
     1321 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
     1381 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
     1441 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     1501 gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag ctggtgatat
     1561 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
     1621 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
     1681 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
     1741 tttcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
     1801 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
     1861 tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc
     1921 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt gatatcgagc
     1981 tcgcttggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
     2041 agaacgctcg gttgccgccg ggcgtttttt attggt
//
