LOCUS       sequence-330206-        2095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for SARS-CoV-2 ORF8 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2095)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2095)
  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-m4046861
FEATURES             Location/Qualifiers
     source          1..2095
                     /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             474..494
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             495..512
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      633..1221
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1122..1141
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1327..1986
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1393..1412)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2057..2091
                     /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 tatgaaattt
      121 cttgttttct taggaatcat cacaactgta gctgcatttc accaagaatg tagtttacag
      181 tcatgtactc aacatcaacc atatgtagtt gatgacccgt gtcctattca cttctattct
      241 aatggtatat tagagtagga gctagaaaat cagcaccttt aattgaattg tgcgtggatg
      301 aggctggttc taaatcaccc attcagtaca tcgatatcgg taattataca gtttcctgtt
      361 taccttttac aattaattgc caggaaccta aattgggtag tcttgtagtg cgttgttcgt
      421 tctatgaaga ctttttagag tatcatgacg ttcgtgttgt tttagatttc atcgagaacc
      481 tgtactttca aagccaccat catcaccatc attaaccggc ttatcggtca gtttcacctg
      541 atttacgtaa aaacccgctt cggcgggttt ttgcttttgg aggggcagaa agatgaatga
      601 ctgtccacga cgctataccc aaaagaaaaa gcttgagatc ctttttttct gcgcgtaatc
      661 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
      721 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
      781 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
      841 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
      901 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
      961 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     1021 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     1081 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     1141 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     1201 ggggggcgga gcctatggaa attgatcggg cacgtaagag gttccaactt tcaccataat
     1261 gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa
     1321 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     1381 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     1441 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     1501 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa
     1561 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     1621 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     1681 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     1741 attgagaata tgtttttcgt ttcagccaat ccctgggtga gtttcaccag ttttgattta
     1801 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     1861 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     1921 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg
     1981 gcgtaatttg atatcgagct cgcttggact cctgttgata gatccagtaa tgacctcaga
     2041 actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggt
//
