LOCUS       sequence-357572-        3650 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for SARS-CoV-2 nsp2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3650)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 3650)
  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-m4046906
FEATURES             Location/Qualifiers
     source          1..3650
                     /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             2000..2020
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2021..2044
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      2165..2753
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2654..2673
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2859..3518
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2925..2944)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3589..3623
                     /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 gtacactcgc tatgtcgata acaacttctg
      121 tggccctgat ggctaccctc ttgagtgcat taaagacctt ctagcacgtg ctggtaaagc
      181 ttcatgcact ttgtccgaac aactggactt tattgacact aagaggggtg tatactgctg
      241 ccgtgaacat gagcatgaaa ttgcttggta cacggaacgt tctgaaaaga gctatgaatt
      301 gcagacacct tttgaaatta aattggcaaa gaaatttgac accttcaatg gggaatgtcc
      361 aaattttgta tttcccttaa attccataat caagactatt caaccaaggg ttgaaaagaa
      421 aaagcttgat ggctttatgg gtagaattcg atctgtctat ccagttgcgt caccaaatga
      481 atgcaaccaa atgtgccttt caactctcat gaagtgtgat cattgtggtg aaacttcatg
      541 gcagacgggc gattttgtta aagccacttg cgaattttgt ggcactgaga atttgactaa
      601 agaaggtgcc actacttgtg gttacttacc ccaaaatgct gttgttaaaa tttattgtcc
      661 agcatgtcac aattcagaag taggacctga gcatagtctt gccgaatacc ataatgaatc
      721 tggcttgaaa accattcttc gtaagggtgg tcgcactatt gcctttggag gctgtgtgtt
      781 ctcttatgtt ggttgccata acaagtgtgc ctattgggtt ccacgtgcta gcgctaacat
      841 aggttgtaac catacaggtg ttgttggaga aggttccgaa ggtcttaatg acaaccttct
      901 tgaaatactc caaaaagaga aagtcaacat caatattgtt ggtgacttta aacttaatga
      961 agagatcgcc attattttgg catctttttc tgcttccaca agtgcttttg tggaaactgt
     1021 gaaaggtttg gattataaag cattcaaaca aattgttgaa tcctgtggta attttaaagt
     1081 tacaaaagga aaagctaaaa aaggtgcctg gaatattggt gaacagaaat caatactgag
     1141 tcctctttat gcatttgcat cagaggctgc tcgtgttgta cgatcaattt tctcccgcac
     1201 tcttgaaact gctcaaaatt ctgtgcgtgt tttacagaag gccgctataa caatactaga
     1261 tggaatttca cagtattcac tgagactcat tgatgctatg atgttcacat ctgatttggc
     1321 tactaacaat ctagttgtaa tggcctacat tacaggtggt gttgttcagt tgacttcgca
     1381 gtggctaact aacatctttg gcactgttta tgaaaaactc aaacccgtcc ttgattggct
     1441 tgaagagaag tttaaggaag gtgtagagtt tcttagagac ggttgggaaa ttgttaaatt
     1501 tatctcaacc tgtgcttgtg aaattgtcgg tggacaaatt gtcacctgtg caaaggaaat
     1561 taaggagagt gttcagacat tctttaagct tgtaaataaa tttttggctt tgtgtgctga
     1621 ctctatcatt attggtggag ctaaacttaa agccttgaat ttaggtgaaa catttgtcac
     1681 gcactcaaag ggattgtaca gaaagtgtgt taaatccaga gaagaaactg gcctactcat
     1741 gcctctaaaa gccccaaaag aaattatctt cttagaggga gaaacacttc ccacagaagt
     1801 gttaacagag gaagttgtct tgaaaactgg tgatttacaa ccattagaac aacctactag
     1861 tgaagctgtt gaagctccat tggttggtac accagtttgt attaacgggc ttatgttgct
     1921 cgaaatcaaa gacacagaaa agtactgtgc ccttgcacct aatatgatgg taacaaacaa
     1981 taccttcaca ctcaaaggcg agaacctgta ctttcaaagc gactacaagg atgatgatga
     2041 caaataaccg gcttatcggt cagtttcacc tgatttacgt aaaaacccgc ttcggcgggt
     2101 ttttgctttt ggaggggcag aaagatgaat gactgtccac gacgctatac ccaaaagaaa
     2161 aagcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2221 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2281 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     2341 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2401 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2461 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2521 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2581 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2641 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2701 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaattgatcg
     2761 ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt
     2821 ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     2881 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     2941 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     3001 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     3061 atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg
     3121 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     3181 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     3241 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtttcagcca
     3301 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     3361 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     3421 cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat
     3481 tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tgatatcgag ctcgcttgga
     3541 ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc
     3601 ggttgccgcc gggcgttttt tattggtcat cgatctcgat cccgcgaaat
//
