LOCUS       sequence-296971-        2096 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 2096)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2096)
  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..2096
                     /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             475..495
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             496..513
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      634..1222
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1123..1142
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1328..1987
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1394..1413)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2058..2092
                     /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 aaatggtata ttagagtagg agctagaaaa tcagcacctt taattgaatt gtgcgtggat
      301 gaggctggtt ctaaatcacc cattcagtac atcgatatcg gtaattatac agtttcctgt
      361 ttacctttta caattaattg ccaggaacct aaattgggta gtcttgtagt gcgttgttcg
      421 ttctatgaag actttttaga gtatcatgac gttcgtgttg ttttagattt catcgagaac
      481 ctgtactttc aaagccacca tcatcaccat cattaaccgg cttatcggtc agtttcacct
      541 gatttacgta aaaacccgct tcggcgggtt tttgcttttg gaggggcaga aagatgaatg
      601 actgtccacg acgctatacc caaaagaaaa agcttgagat cctttttttc tgcgcgtaat
      661 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
      721 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
      781 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
      841 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
      901 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
      961 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     1021 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     1081 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     1141 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     1201 aggggggcgg agcctatgga aattgatcgg gcacgtaaga ggttccaact ttcaccataa
     1261 tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga
     1321 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
     1381 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
     1441 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
     1501 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa
     1561 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
     1621 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
     1681 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
     1741 tattgagaat atgtttttcg tttcagccaa tccctgggtg agtttcacca gttttgattt
     1801 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
     1861 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg
     1921 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     1981 ggcgtaattt gatatcgagc tcgcttggac tcctgttgat agatccagta atgacctcag
     2041 aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggt
//
