LOCUS       sequence-287863-        2559 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 2559)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2559)
  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-m4046727
FEATURES             Location/Qualifiers
     source          1..2559
                     /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..980
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      1097..1685
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1586..1605
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1791..2450
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1857..1876)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2521..2555
                     /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 agctacccgt
      961 atgacgttcc tgattatgcc taaccggctt atcggtcagt ttcacctgat ttacgtaaaa
     1021 acccgcttcg gcgggttttt gcttttggag gggcagaaag atgaatgact gtccacgacg
     1081 ctatacccaa aagaaattga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1141 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1201 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1261 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1321 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1381 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1441 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1501 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1561 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1621 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1681 ggaaattgat cgggcacgta agaggttcca actttcacca taatgaaata agatcactac
     1741 cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa
     1801 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
     1861 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct
     1921 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
     1981 cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt gagctggtga
     2041 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat
     2101 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg
     2161 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt
     2221 tcgtttcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg
     2281 acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc
     2341 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
     2401 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttgatatcg
     2461 agctcgcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
     2521 ttcagaacgc tcggttgccg ccgggcgttt tttattggt
//
