LOCUS       sequence-296909-        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-m4046800
FEATURES             Location/Qualifiers
     source          1..2558
                     /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..971
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      1092..1680
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1581..1600
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1786..2445
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1852..1871)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2516..2550
                     /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 gacttgttta
      121 tgcgtatttt cacaatcggt acggtgaccc tcaaacaggg cgagatcaag gatgcgaccc
      181 caagtgactt cgttcgcgct actgccacta ttcccatcca agcatctctg ccttttgggt
      241 ggcttatcgt aggtgttgct ctgctggcag tcttccagtc ggcgagcaaa ataatcaccc
      301 taaaaaagcg ttggcagctg gctctgtcta aaggcgtaca tttcgtctgc aacctgcttt
      361 tactgttcgt tactgtgtat tcccacctgt tgctggttgc cgctggactg gaagcaccgt
      421 ttctgtactt atacgctctg gtttatttct tgcagagcat caattttgtg cggataatta
      481 tgcgcctgtg gctgtgttgg aaatgccgtt caaaaaaccc gctgctttac gatgcgaact
      541 acttcttatg ctggcacacc aactgttacg attattgcat tccgtataat tccgtaacca
      601 gctctatcgt tatcacgagc ggcgacggca ctacttcccc aatcagtgaa catgactacc
      661 agattggtgg ttacactgaa aaatgggagt ctggcgttaa agattgtgtg gtactgcact
      721 cctatttcac gtctgactac taccaactgt acagcaccca gctgtcgacc gacacaggtg
      781 tcgaacatgt cacttttttc atctacaaca agatcgttga tgaaccggag gaacacgttc
      841 agattcacac cattgatgga tcatctggtg tggttaaccc ggtgatggaa ccgatctatg
      901 acgaacccac gacaaccact tctgtacctc tggagaacct gtactttcaa agccaccatc
      961 atcaccatca ttaaccggct tatcggtcag tttcacctga tttacgtaaa aacccgcttc
     1021 ggcgggtttt tgcttttgga ggggcagaaa gatgaatgac tgtccacgac gctataccca
     1081 aaagaaaaag cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1141 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     1201 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     1261 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     1321 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     1381 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     1441 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     1501 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     1561 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     1621 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     1681 ttgatcgggc acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc
     1741 gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
     1801 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     1861 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     1921 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
     1981 ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag acggtgagct ggtgatatgg
     2041 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
     2101 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
     2161 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtt
     2221 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     2281 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     2341 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
     2401 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaatttga tatcgagctc
     2461 gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg atttgttcag
     2521 aacgctcggt tgccgccggg cgttttttat tggtcatc
//
