LOCUS       sequence-287824-        2640 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression plasmid for SARS-CoV-2 3C-like proteinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2640)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2640)
  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-m4046754
FEATURES             Location/Qualifiers
     source          1..2640
                     /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             1023..1043
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1044..1061
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      1178..1766
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1667..1686
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1872..2531
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1938..1957)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2602..2636
                     /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 ggttttagaa
      121 aaatggcatt cccatctggt aaagttgagg gttgtatggt acaagtaact tgtggtacaa
      181 ctacacttaa cggtctttgg cttgatgacg tagtttactg tccaagacat gtgatctgca
      241 cctctgaaga catgcttaac cctaattatg aagatttact cattcgtaag tctaatcata
      301 atttcttggt acaggctggt aatgttcaac tcagggttat tggacattct atgcaaaatt
      361 gtgtacttaa gcttaaggtt gatacagcca atcctaagac acctaagtat aagtttgttc
      421 gcattcaacc aggacagact ttttcagtgt tagcttgtta caatggttca ccatctggtg
      481 tttaccaatg tgctatgagg cccaatttca ctattaaggg ttcattcctt aatggttcat
      541 gtggtagtgt tggttttaac atagattatg actgtgtctc tttttgttac atgcaccata
      601 tggaattacc aactggagtt catgctggca cagacttaga aggtaacttt tatggacctt
      661 ttgttgacag gcaaacagca caagcagctg gtacggacac aactattaca gttaatgttt
      721 tagcttggtt gtacgctgct gttataaatg gagacaggtg gtttctcaat cgatttacca
      781 caactcttaa tgactttaac cttgtggcta tgaagtacaa ttatgaacct ctaacacaag
      841 accatgttga catactagga cctctttctg ctcaaactgg aattgccgtt ttagatatgt
      901 gtgcttcatt aaaagaatta ctgcaaaatg gtatgaatgg acgtaccata ttgggtagtg
      961 ctttattaga agatgaattt acaccttttg atgttgttag acaatgctca ggtgttactt
     1021 tcgagaacct gtactttcaa agccaccatc atcaccatca ttaaccggct tatcggtcag
     1081 tttcacctga tttacgtaaa aacccgcttc ggcgggtttt tgcttttgga ggggcagaaa
     1141 gatgaatgac tgtccacgac gctataccca aaagaaattg agatcctttt tttctgcgcg
     1201 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1261 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1321 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1381 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1441 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     1501 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     1561 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     1621 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     1681 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     1741 cgtcaggggg gcggagccta tggaaattga tcgggcacgt aagaggttcc aactttcacc
     1801 ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta
     1861 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
     1921 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
     1981 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
     2041 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattt cgtatggcaa
     2101 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
     2161 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
     2221 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
     2281 ggtttattga gaatatgttt ttcgtttcag ccaatccctg ggtgagtttc accagttttg
     2341 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt
     2401 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
     2461 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
     2521 gcggggcgta atttgatatc gagctcgctt ggactcctgt tgatagatcc agtaatgacc
     2581 tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
//
