LOCUS       sequence-292485-        2558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 ORF3a protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2558)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2558)
  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-m4046959
FEATURES             Location/Qualifiers
     source          1..2558
                     /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             937..957
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             958..975
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      1096..1684
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1585..1604
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1790..2449
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1856..1875)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2520..2554
                     /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 tatggatttg
      121 tttatgagaa tcttcacaat tggaactgta actttgaagc aaggtgaaat caaggatgct
      181 actccttcag attttgttcg cgctactgca acgataccga tacaagcctc actccctttc
      241 ggatggctta ttgttggcgt tgcacttctt gctgtttttc agagcgcttc caaaatcata
      301 accctcaaaa agagatggca actagcactc tccaagggtg ttcactttgt ttgcaacttg
      361 ctgttgttgt ttgtaacagt ttactcacac cttttgctcg ttgctgctgg ccttgaagcc
      421 ccttttctct atctttatgc tttagtctac ttcttgcaga gtataaactt tgtaagaata
      481 ataatgaggc tttggctttg ctggaaatgc cgttccaaaa acccattact ttatgatgcc
      541 aactattttc tttgctggca tactaattgt tacgactatt gtatacctta caatagtgta
      601 acttcttcaa ttgtcattac ttcaggtgat ggcacaacaa gtcctatttc tgaacatgac
      661 taccagattg gtggttatac tgaaaaatgg gaatctggag taaaagactg tgttgtatta
      721 cacagttact tcacttcaga ctattaccag ctgtactcaa ctcaattgag tacagacact
      781 ggtgttgaac atgttacctt cttcatctac aataaaattg ttgatgagcc tgaagaacat
      841 gtccaaattc acacaatcga cggttcatcc ggagttgtta atccagtaat ggaaccaatt
      901 tatgatgaac cgacgacgac tactagcgtg cctttggaga acctgtactt tcaaagccac
      961 catcatcacc atcattaacc ggcttatcgg tcagtttcac ctgatttacg taaaaacccg
     1021 cttcggcggg tttttgcttt tggaggggca gaaagatgaa tgactgtcca cgacgctata
     1081 cccaaaagaa aaagcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1141 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1201 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     1261 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1321 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1381 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1441 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1501 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1561 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     1621 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     1681 gaaattgatc gggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc
     1741 gggcgtattt tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     1801 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     1861 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     1921 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     1981 ccgcctgatg aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat
     2041 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     2101 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     2161 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     2221 cgtttcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     2281 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     2341 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     2401 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttgatatcga
     2461 gctcgcttgg actcctgttg atagatccag taatgacctc agaactccat ctggatttgt
     2521 tcagaacgct cggttgccgc cgggcgtttt ttattggt
//
