LOCUS       sequence-298359-        2564 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 2564)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2564)
  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-m4046751
FEATURES             Location/Qualifiers
     source          1..2564
                     /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             933..953
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             954..977
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     rep_origin      1098..1686
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1587..1606
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1792..2451
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1858..1877)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2522..2556
                     /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 gatttgttta
      121 tgagaatctt cacaattgga actgtaactt tgaagcaagg tgaaatcaag gatgctactc
      181 cttcagattt tgttcgcgct actgcaacga taccgataca agcctcactc cctttcggat
      241 ggcttattgt tggcgttgca cttcttgctg tttttcagag cgcttccaaa atcataaccc
      301 tcaaaaagag atggcaacta gcactctcca agggtgttca ctttgtttgc aacttgctgt
      361 tgttgtttgt aacagtttac tcacaccttt tgctcgttgc tgctggcctt gaagcccctt
      421 ttctctatct ttatgcttta gtctacttct tgcagagtat aaactttgta agaataataa
      481 tgaggctttg gctttgctgg aaatgccgtt ccaaaaaccc attactttat gatgccaact
      541 attttctttg ctggcatact aattgttacg actattgtat accttacaat agtgtaactt
      601 cttcaattgt cattacttca ggtgatggca caacaagtcc tatttctgaa catgactacc
      661 agattggtgg ttatactgaa aaatgggaat ctggagtaaa agactgtgtt gtattacaca
      721 gttacttcac ttcagactat taccagctgt actcaactca attgagtaca gacactggtg
      781 ttgaacatgt taccttcttc atctacaata aaattgttga tgagcctgaa gaacatgtcc
      841 aaattcacac aatcgacggt tcatccggag ttgttaatcc agtaatggaa ccaatttatg
      901 atgaaccgac gacgactact agcgtgcctt tggagaacct gtactttcaa agctggagcc
      961 atccgcagtt tgaaaaataa ccggcttatc ggtcagtttc acctgattta cgtaaaaacc
     1021 cgcttcggcg ggtttttgct tttggagggg cagaaagatg aatgactgtc cacgacgcta
     1081 tacccaaaag aaaaagcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1141 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1201 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1261 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1321 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1381 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1441 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1501 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1561 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1621 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     1681 tggaaattga tcgggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta
     1741 ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa aatggagaaa
     1801 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag
     1861 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc
     1921 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
     1981 gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg
     2041 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
     2101 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
     2161 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
     2221 ttcgtttcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     2281 gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg
     2341 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga
     2401 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta atttgatatc
     2461 gagctcgctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc atctggattt
     2521 gttcagaacg ctcggttgcc gccgggcgtt ttttattggt catc
//
