LOCUS       sequence-325546-        1922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 ORF6 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1922)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 1922)
  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-m4046744
FEATURES             Location/Qualifiers
     source          1..1922
                     /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             291..311
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             312..335
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      456..1044
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     945..964
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1150..1809
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1216..1235)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1880..1914
                     /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 tttcatctcg
      121 ttgactttca ggttactata gcagagatat tactaattat tatgaggact tttaaagttt
      181 ccatttggaa tcttgattac atcataaacc tcataattaa aaatttatct aagtcactaa
      241 ctgagaataa atattctcaa ttagatgaag agcaaccaat ggagattgat gagaacctgt
      301 actttcaaag cgactacaag gatgatgatg acaaataacc ggcttatcgg tcagtttcac
      361 ctgatttacg taaaaacccg cttcggcggg tttttgcttt tggaggggca gaaagatgaa
      421 tgactgtcca cgacgctata cccaaaagaa aaagcttgag atcctttttt tctgcgcgta
      481 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
      541 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
      601 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
      661 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
      721 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
      781 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
      841 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
      901 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
      961 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     1021 tcaggggggc ggagcctatg gaaattgatc gggcacgtaa gaggttccaa ctttcaccat
     1081 aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc aggagctaag
     1141 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     1201 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     1261 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     1321 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaatttcg tatggcaatg
     1381 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     1441 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     1501 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     1561 tttattgaga atatgttttt cgtttcagcc aatccctggg tgagtttcac cagttttgat
     1621 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     1681 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     1741 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc
     1801 ggggcgtaat ttgatatcga gctcgcttgg actcctgttg atagatccag taatgacctc
     1861 agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtca
     1921 tc
//
