LOCUS       sequence-357573-        3644 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 3644)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 3644)
  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-m4046879
FEATURES             Location/Qualifiers
     source          1..3644
                     /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..2038
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      2159..2747
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2648..2667
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2853..3512
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2919..2938)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3583..3617
                     /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 caccatcatc accatcatta
     2041 accggcttat cggtcagttt cacctgattt acgtaaaaac ccgcttcggc gggtttttgc
     2101 ttttggaggg gcagaaagat gaatgactgt ccacgacgct atacccaaaa gaaaaagctt
     2161 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2221 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2281 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     2341 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2401 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2461 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2521 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2581 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2641 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2701 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaattg atcgggcacg
     2761 taagaggttc caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt
     2821 tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca
     2881 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
     2941 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
     3001 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
     3061 atccggaatt tcgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc
     3121 cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc
     3181 acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa
     3241 acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtttca gccaatccct
     3301 gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg
     3361 ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc
     3421 aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac
     3481 agtactgcga tgagtggcag ggcggggcgt aatttgatat cgagctcgct tggactcctg
     3541 ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc
     3601 cgccgggcgt tttttattgg tcatcgatct cgatcccgcg aaat
//
