LOCUS       sequence-292503-        2600 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 2600)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2600)
  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-m4046942
FEATURES             Location/Qualifiers
     source          1..2600
                     /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             956..976
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             977..994
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      1115..1703
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1604..1623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1809..2468
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1875..1894)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2539..2573
                     /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 ggcagtgaaa agaacaatca agggtacaca
      121 ccactggttg ttactcacaa ttttgacttc acttttagtt ttagtccaga gtactcaatg
      181 gtctttgttc ttttttttgt atgaaaatgc ctttttacct tttgctatgg gtattattgc
      241 tatgtctgct tttgcaatga tgtttgtcaa acataagcat gcatttctct gtttgttttt
      301 gttaccttct cttgccactg tagcttattt taatatggtc tatatgcctg ctagttgggt
      361 gatgcgtatt atgacatggt tggatatggt tgatactagt ttgtctggtt ttaagctaaa
      421 agactgtgtt atgtatgcat cagctgtagt gttactaatc cttatgacag caagaactgt
      481 gtatgatgat ggtgctagga gagtgtggac acttatgaat gtcttgacac tcgtttataa
      541 agtttattat ggtaatgctt tagatcaagc catttccatg tgggctctta taatctctgt
      601 tacttctaac tactcaggtg tagttacaac tgtcatgttt ttggccagag gtattgtttt
      661 tatgtgtgtt gagtattgcc ctattttctt cataactggt aatacacttc agtgtataat
      721 gctagtttat tgtttcttag gctatttttg tacttgttac tttggcctct tttgtttact
      781 caaccgctac tttagactga ctcttggtgt ttatgattac ttagtttcta cacaggagtt
      841 tagatatatg aattcacagg gactactccc acccaagaat agcatagatg ccttcaaact
      901 caacattaaa ttgttgggtg ttggtggcaa accttgtatc aaagtagcca ctgtagagaa
      961 cctgtacttt caaagccacc atcatcacca tcattaaccg gcttatcggt cagtttcacc
     1021 tgatttacgt aaaaacccgc ttcggcgggt ttttgctttt ggaggggcag aaagatgaat
     1081 gactgtccac gacgctatac ccaaaagaaa aagcttgaga tccttttttt ctgcgcgtaa
     1141 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     1201 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     1261 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     1321 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     1381 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     1441 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     1501 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     1561 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     1621 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     1681 caggggggcg gagcctatgg aaattgatcg ggcacgtaag aggttccaac tttcaccata
     1741 atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg
     1801 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
     1861 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
     1921 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
     1981 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaatttcgt atggcaatga
     2041 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
     2101 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
     2161 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
     2221 ttattgagaa tatgtttttc gtttcagcca atccctgggt gagtttcacc agttttgatt
     2281 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
     2341 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
     2401 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
     2461 gggcgtaatt tgatatcgag ctcgcttgga ctcctgttga tagatccagt aatgacctca
     2521 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtcat
     2581 cgatctcgat cccgcgaaat
//
