LOCUS       sequence-287804-        2598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 nsp6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2598)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2598)
  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-m4046916
FEATURES             Location/Qualifiers
     source          1..2598
                     /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             981..1001
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1002..1019
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      1136..1724
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1625..1644
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1830..2489
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1896..1915)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2560..2594
                     /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 tccgcggtga
      121 agcgtacgat taaaggcact caccattggc tgttacttac aatcctaacc agcctgctgg
      181 tcctcgttca aagtactcag tggtcgctgt tctttttctt gtatgagaac gccttcctgc
      241 cgtttgcaat ggggatcata gctatgtcag cgttcgctat gatgtttgtt aaacacaaac
      301 atgcattctt atgtctgttt ctgctgccaa gcctggctac cgtagcatac ttcaatatgg
      361 tttacatgcc ggcttcctgg gtgatgcgca ttatgacttg gctggatatg gtagacacct
      421 ctctgtctgg tttcaagctt aaagactgcg ttatgtatgc cagcgcggtg gttctgctga
      481 tcctgatgac cgctaggacc gtctacgatg atggtgctcg tcgcgtatgg actctaatga
      541 acgttctgac tcttgtatac aaagtctatt acggcaacgc actggaccag gcgatctcta
      601 tgtgggccct gatcatttct gttacatcca attacagtgg tgtagttacc acggtaatgt
      661 tccttgcccg tggcattgtg ttcatgtgcg tggaatattg ccctatcttc tttatcactg
      721 gtaacaccct gcaatgtatc atgttggttt actgctttct gggctacttt tgcacttgct
      781 atttcgggct gttctgttta ctcaaccgtt actttcgcct gaccctgggt gtctacgact
      841 acctggttag cacacaggaa tttcgctaca tgaacagcca gggactgctt cccccgaaaa
      901 attctattga tgctttcaaa ctgaacatca agttgttggg cgtgggtggc aaaccgtgca
      961 tcaaagtggc gacggttcag gagaacctgt actttcaaag ccaccatcat caccatcatt
     1021 aaccggctta tcggtcagtt tcacctgatt tacgtaaaaa cccgcttcgg cgggtttttg
     1081 cttttggagg ggcagaaaga tgaatgactg tccacgacgc tatacccaaa agaaattgag
     1141 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1201 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1261 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     1321 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1381 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1441 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1501 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     1561 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     1621 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     1681 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaattgatc gggcacgtaa
     1741 gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat
     1801 cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg
     1861 ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat
     1921 gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa
     1981 ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc
     2041 cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt
     2101 gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg
     2161 acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc
     2221 tggcctattt ccctaaaggg tttattgaga atatgttttt cgtttcagcc aatccctggg
     2281 tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt
     2341 tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg
     2401 ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt
     2461 actgcgatga gtggcagggc ggggcgtaat ttgatatcga gctcgcttgg actcctgttg
     2521 atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc
     2581 cgggcgtttt ttattggt
//
