LOCUS       sequence-382441-        2570 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 2570)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2570)
  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-m4046882
FEATURES             Location/Qualifiers
     source          1..2570
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             914..934
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             935..964
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     rep_origin      1085..1673
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1574..1593
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1779..2438
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1845..1864)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2509..2543
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag actcttctat ggatttgttt atgagaatct tcacaattgg
      121 aactgtaact ttgaagcaag gtgaaatcaa ggatgctact ccttcagatt ttgttcgcgc
      181 tactgcaacg ataccgatac aagcctcact ccctttcgga tggcttattg ttggcgttgc
      241 acttcttgct gtttttcaga gcgcttccaa aatcataacc ctcaaaaaga gatggcaact
      301 agcactctcc aagggtgttc actttgtttg caacttgctg ttgttgtttg taacagttta
      361 ctcacacctt ttgctcgttg ctgctggcct tgaagcccct tttctctatc tttatgcttt
      421 agtctacttc ttgcagagta taaactttgt aagaataata atgaggcttt ggctttgctg
      481 gaaatgccgt tccaaaaacc cattacttta tgatgccaac tattttcttt gctggcatac
      541 taattgttac gactattgta taccttacaa tagtgtaact tcttcaattg tcattacttc
      601 aggtgatggc acaacaagtc ctatttctga acatgactac cagattggtg gttatactga
      661 aaaatgggaa tctggagtaa aagactgtgt tgtattacac agttacttca cttcagacta
      721 ttaccagctg tactcaactc aattgagtac agacactggt gttgaacatg ttaccttctt
      781 catctacaat aaaattgttg atgagcctga agaacatgtc caaattcaca caatcgacgg
      841 ttcatccgga gttgttaatc cagtaatgga accaatttat gatgaaccga cgacgactac
      901 tagcgtgcct ttggagaacc tgtactttca aagcgagcag aagcttattt ccgaagaaga
      961 tctctaaccg gcttatcggt cagtttcacc tgatttacgt aaaaacccgc ttcggcgggt
     1021 ttttgctttt ggaggggcag aaagatgaat gactgtccac gacgctatac ccaaaagaaa
     1081 aagcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1141 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1201 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1261 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1321 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1381 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     1441 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1501 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1561 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1621 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaattgatcg
     1681 ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt
     1741 ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     1801 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     1861 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     1921 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     1981 atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg
     2041 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     2101 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     2161 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtttcagcca
     2221 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     2281 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     2341 cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat
     2401 tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tgatatcgag ctcgcttgga
     2461 ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc
     2521 ggttgccgcc gggcgttttt tattggtcat cgatctcgat cccgcgaaat
//
