LOCUS       sequence-323301-        2075 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 2075)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2075)
  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-m4046914
FEATURES             Location/Qualifiers
     source          1..2075
                     /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             425..445
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             446..469
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     rep_origin      590..1178
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1079..1098
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1284..1943
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1350..1369)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2014..2048
                     /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 gaatgagctt agtcctgttg cactacgaca
      121 gatgtcttgt gctgccggta ctacacaaac tgcttgcact gatgacaatg cgttagctta
      181 ctacaacaca acaaagggag gtaggtttgt acttgcactg ttatccgatt tacaggattt
      241 gaaatgggct agattcccta agagtgatgg aactggtact atctatacag aactggaacc
      301 accttgtagg tttgttacag acacacctaa aggtcctaaa gtgaagtatt tatactttat
      361 taaaggatta aacaacctaa atagaggtat ggtacttggt agtttagctg ccacagtacg
      421 tctagagaac ctgtactttc aaagctggag ccatccgcag tttgaaaaat aaccggctta
      481 tcggtcagtt tcacctgatt tacgtaaaaa cccgcttcgg cgggtttttg cttttggagg
      541 ggcagaaaga tgaatgactg tccacgacgc tatacccaaa agaaaaagct tgagatcctt
      601 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
      661 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
      721 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
      781 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
      841 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
      901 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
      961 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     1021 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     1081 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     1141 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaatt gatcgggcac gtaagaggtt
     1201 ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat
     1261 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
     1321 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
     1381 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     1441 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     1501 ttcgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
     1561 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
     1621 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
     1681 atttccctaa agggtttatt gagaatatgt ttttcgtttc agccaatccc tgggtgagtt
     1741 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     1801 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     1861 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
     1921 atgagtggca gggcggggcg taatttgata tcgagctcgc ttggactcct gttgatagat
     1981 ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg
     2041 ttttttattg gtcatcgatc tcgatcccgc gaaat
//
