LOCUS       sequence-358322-        3656 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 3656)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 3656)
  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-m4046907
FEATURES             Location/Qualifiers
     source          1..3656
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1..23
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1942..1962
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1963..1992
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     rep_origin      2113..2701
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2602..2621
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2807..3466
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2873..2892)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3537..3571
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     primer_bind     3599..3618
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3599..3617
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3618..3642
                     /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)."
ORIGIN
        1 tttgtttaac tttaagaagg agactcttct atgtacactc gctatgtcga taacaacttc
       61 tgtggccctg atggctaccc tcttgagtgc attaaagacc ttctagcacg tgctggtaaa
      121 gcttcatgca ctttgtccga acaactggac tttattgaca ctaagagggg tgtatactgc
      181 tgccgtgaac atgagcatga aattgcttgg tacacggaac gttctgaaaa gagctatgaa
      241 ttgcagacac cttttgaaat taaattggca aagaaatttg acaccttcaa tggggaatgt
      301 ccaaattttg tatttccctt aaattccata atcaagacta ttcaaccaag ggttgaaaag
      361 aaaaagcttg atggctttat gggtagaatt cgatctgtct atccagttgc gtcaccaaat
      421 gaatgcaacc aaatgtgcct ttcaactctc atgaagtgtg atcattgtgg tgaaacttca
      481 tggcagacgg gcgattttgt taaagccact tgcgaatttt gtggcactga gaatttgact
      541 aaagaaggtg ccactacttg tggttactta ccccaaaatg ctgttgttaa aatttattgt
      601 ccagcatgtc acaattcaga agtaggacct gagcatagtc ttgccgaata ccataatgaa
      661 tctggcttga aaaccattct tcgtaagggt ggtcgcacta ttgcctttgg aggctgtgtg
      721 ttctcttatg ttggttgcca taacaagtgt gcctattggg ttccacgtgc tagcgctaac
      781 ataggttgta accatacagg tgttgttgga gaaggttccg aaggtcttaa tgacaacctt
      841 cttgaaatac tccaaaaaga gaaagtcaac atcaatattg ttggtgactt taaacttaat
      901 gaagagatcg ccattatttt ggcatctttt tctgcttcca caagtgcttt tgtggaaact
      961 gtgaaaggtt tggattataa agcattcaaa caaattgttg aatcctgtgg taattttaaa
     1021 gttacaaaag gaaaagctaa aaaaggtgcc tggaatattg gtgaacagaa atcaatactg
     1081 agtcctcttt atgcatttgc atcagaggct gctcgtgttg tacgatcaat tttctcccgc
     1141 actcttgaaa ctgctcaaaa ttctgtgcgt gttttacaga aggccgctat aacaatacta
     1201 gatggaattt cacagtattc actgagactc attgatgcta tgatgttcac atctgatttg
     1261 gctactaaca atctagttgt aatggcctac attacaggtg gtgttgttca gttgacttcg
     1321 cagtggctaa ctaacatctt tggcactgtt tatgaaaaac tcaaacccgt ccttgattgg
     1381 cttgaagaga agtttaagga aggtgtagag tttcttagag acggttggga aattgttaaa
     1441 tttatctcaa cctgtgcttg tgaaattgtc ggtggacaaa ttgtcacctg tgcaaaggaa
     1501 attaaggaga gtgttcagac attctttaag cttgtaaata aatttttggc tttgtgtgct
     1561 gactctatca ttattggtgg agctaaactt aaagccttga atttaggtga aacatttgtc
     1621 acgcactcaa agggattgta cagaaagtgt gttaaatcca gagaagaaac tggcctactc
     1681 atgcctctaa aagccccaaa agaaattatc ttcttagagg gagaaacact tcccacagaa
     1741 gtgttaacag aggaagttgt cttgaaaact ggtgatttac aaccattaga acaacctact
     1801 agtgaagctg ttgaagctcc attggttggt acaccagttt gtattaacgg gcttatgttg
     1861 ctcgaaatca aagacacaga aaagtactgt gcccttgcac ctaatatgat ggtaacaaac
     1921 aataccttca cactcaaagg cgagaacctg tactttcaaa gcgagcagaa gcttatttcc
     1981 gaagaagatc tctaaccggc ttatcggtca gtttcacctg atttacgtaa aaacccgctt
     2041 cggcgggttt ttgcttttgg aggggcagaa agatgaatga ctgtccacga cgctataccc
     2101 aaaagaaaaa gcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2161 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2221 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2281 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2341 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2401 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2461 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2521 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2581 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2641 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2701 attgatcggg cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg
     2761 cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat
     2821 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt
     2881 tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt
     2941 aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg
     3001 cctgatgaat gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg
     3061 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct
     3121 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc
     3181 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt
     3241 ttcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa
     3301 cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat
     3361 gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct
     3421 taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttg atatcgagct
     3481 cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca
     3541 gaacgctcgg ttgccgccgg gcgtttttta ttggtcatcg atctcgatcc cgcgaaatta
     3601 atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga aataat
//
