LOCUS       sequence-297009-        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-m4046848
FEATURES             Location/Qualifiers
     source          1..3650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..43
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        24..42
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    43..67
                     /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             82..104
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2023..2043
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2044..2067
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     rep_origin      2188..2776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2677..2696
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2882..3541
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2948..2967)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3612..3646
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
ORIGIN
        1 catcgatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
       61 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagactcttc tatgtacact
      121 cgctatgtcg ataacaactt ctgtggccct gatggctacc ctcttgagtg cattaaagac
      181 cttctagcac gtgctggtaa agcttcatgc actttgtccg aacaactgga ctttattgac
      241 actaagaggg gtgtatactg ctgccgtgaa catgagcatg aaattgcttg gtacacggaa
      301 cgttctgaaa agagctatga attgcagaca ccttttgaaa ttaaattggc aaagaaattt
      361 gacaccttca atggggaatg tccaaatttt gtatttccct taaattccat aatcaagact
      421 attcaaccaa gggttgaaaa gaaaaagctt gatggcttta tgggtagaat tcgatctgtc
      481 tatccagttg cgtcaccaaa tgaatgcaac caaatgtgcc tttcaactct catgaagtgt
      541 gatcattgtg gtgaaacttc atggcagacg ggcgattttg ttaaagccac ttgcgaattt
      601 tgtggcactg agaatttgac taaagaaggt gccactactt gtggttactt accccaaaat
      661 gctgttgtta aaatttattg tccagcatgt cacaattcag aagtaggacc tgagcatagt
      721 cttgccgaat accataatga atctggcttg aaaaccattc ttcgtaaggg tggtcgcact
      781 attgcctttg gaggctgtgt gttctcttat gttggttgcc ataacaagtg tgcctattgg
      841 gttccacgtg ctagcgctaa cataggttgt aaccatacag gtgttgttgg agaaggttcc
      901 gaaggtctta atgacaacct tcttgaaata ctccaaaaag agaaagtcaa catcaatatt
      961 gttggtgact ttaaacttaa tgaagagatc gccattattt tggcatcttt ttctgcttcc
     1021 acaagtgctt ttgtggaaac tgtgaaaggt ttggattata aagcattcaa acaaattgtt
     1081 gaatcctgtg gtaattttaa agttacaaaa ggaaaagcta aaaaaggtgc ctggaatatt
     1141 ggtgaacaga aatcaatact gagtcctctt tatgcatttg catcagaggc tgctcgtgtt
     1201 gtacgatcaa ttttctcccg cactcttgaa actgctcaaa attctgtgcg tgttttacag
     1261 aaggccgcta taacaatact agatggaatt tcacagtatt cactgagact cattgatgct
     1321 atgatgttca catctgattt ggctactaac aatctagttg taatggccta cattacaggt
     1381 ggtgttgttc agttgacttc gcagtggcta actaacatct ttggcactgt ttatgaaaaa
     1441 ctcaaacccg tccttgattg gcttgaagag aagtttaagg aaggtgtaga gtttcttaga
     1501 gacggttggg aaattgttaa atttatctca acctgtgctt gtgaaattgt cggtggacaa
     1561 attgtcacct gtgcaaagga aattaaggag agtgttcaga cattctttaa gcttgtaaat
     1621 aaatttttgg ctttgtgtgc tgactctatc attattggtg gagctaaact taaagccttg
     1681 aatttaggtg aaacatttgt cacgcactca aagggattgt acagaaagtg tgttaaatcc
     1741 agagaagaaa ctggcctact catgcctcta aaagccccaa aagaaattat cttcttagag
     1801 ggagaaacac ttcccacaga agtgttaaca gaggaagttg tcttgaaaac tggtgattta
     1861 caaccattag aacaacctac tagtgaagct gttgaagctc cattggttgg tacaccagtt
     1921 tgtattaacg ggcttatgtt gctcgaaatc aaagacacag aaaagtactg tgcccttgca
     1981 cctaatatga tggtaacaaa caataccttc acactcaaag gcgagaacct gtactttcaa
     2041 agctggagcc atccgcagtt tgaaaaataa ccggcttatc ggtcagtttc acctgattta
     2101 cgtaaaaacc cgcttcggcg ggtttttgct tttggagggg cagaaagatg aatgactgtc
     2161 cacgacgcta tacccaaaag aaaaagcttg agatcctttt tttctgcgcg taatctgctg
     2221 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2281 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     2341 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2401 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2461 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2521 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2581 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2641 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2701 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2761 gcggagccta tggaaattga tcgggcacgt aagaggttcc aactttcacc ataatgaaat
     2821 aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa
     2881 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
     2941 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
     3001 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
     3061 tcacattctt gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg
     3121 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
     3181 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
     3241 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
     3301 gaatatgttt ttcgtttcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     3361 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
     3421 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
     3481 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     3541 atttgatatc gagctcgctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
     3601 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
//
