LOCUS       sequence-307625-        2679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for SARS-CoV-2 nsp3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2679)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2679)
  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-m4095421
FEATURES             Location/Qualifiers
     source          1..2679
                     /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             1062..1082
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1083..1100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      1217..1805
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1706..1725
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1911..2570
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1977..1996)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2641..2675
                     /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 gaagtgagga
      121 ctattaaggt gtttacaaca gtagacaaca ttaacctcca cacgcaagtt gtggacatgt
      181 caatgacata tggacaacag tttggtccaa cttatttgga tggagctgat gttactaaaa
      241 taaaacctca taattcacat gaaggtaaaa cattttatgt tttacctaat gatgacactc
      301 tacgtgttga ggcttttgag tactaccaca caactgatcc tagttttctg ggtaggtaca
      361 tgtcagcatt aaatcacact aaaaagtgga aatacccaca agttaatggt ttaacttcta
      421 ttaaatgggc agataacaac tgttatcttg ccactgcatt gttaacactc caacaaatag
      481 agttgaagtt taatccaccg gctctacaag atgcttatta cagagcaagg gctggtgaag
      541 ctgctaactt ttgtgcactt atcttagcct actgtaataa gacagtaggt gagttaggtg
      601 atgttagaga aacaatgagt tacttgtttc aacatgccaa tttagattct tgcaaaagag
      661 tcttgaacgt ggtgtgtaaa acttgtggac aacagcagac aacccttaag ggtgtagaag
      721 ctgttatgta catgggcaca ctttcttatg aacaatttaa gaaaggtgtt cagatacctt
      781 gtacgtgtgg taaacaagct acaaaatatc tagtacaaca ggagtcacct tttgttatga
      841 tgtcagcacc acccgctcag tatgaactta agcatggtac atttacttgt gctagtgagt
      901 acactggtaa ttaccaatgt ggtcactata aacatataac ttctaaagaa actttgtatt
      961 gcatagacgg tgctttactt acaaagtcct cagaatacaa aggtcctatt acggatgttt
     1021 tctacaaaga aaacagttac acaacaacca taaaaccagt tgagaacctg tactttcaaa
     1081 gccaccatca tcaccatcat taaccggctt atcggtcagt ttcacctgat ttacgtaaaa
     1141 acccgcttcg gcgggttttt gcttttggag gggcagaaag atgaatgact gtccacgacg
     1201 ctatacccaa aagaaattga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1261 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1321 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1381 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1441 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1501 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1561 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1621 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1681 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1741 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1801 ggaaattgat cgggcacgta agaggttcca actttcacca taatgaaata agatcactac
     1861 cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa
     1921 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
     1981 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct
     2041 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
     2101 cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt gagctggtga
     2161 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat
     2221 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg
     2281 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt
     2341 tcgtttcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg
     2401 acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc
     2461 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
     2521 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttgatatcg
     2581 agctcgcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
     2641 ttcagaacgc tcggttgccg ccgggcgttt tttattggt
//
