LOCUS       sequence-287828-        2646 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 2646)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2646)
  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-m4046922
FEATURES             Location/Qualifiers
     source          1..2646
                     /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..1067
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      1184..1772
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1673..1692
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1878..2537
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1944..1963)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2608..2642
                     /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 agcgactaca aggatgatga tgacaaataa ccggcttatc
     1081 ggtcagtttc acctgattta cgtaaaaacc cgcttcggcg ggtttttgct tttggagggg
     1141 cagaaagatg aatgactgtc cacgacgcta tacccaaaag aaattgagat cctttttttc
     1201 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     1261 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     1321 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     1381 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     1441 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     1501 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     1561 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     1621 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     1681 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     1741 gatgctcgtc aggggggcgg agcctatgga aattgatcgg gcacgtaaga ggttccaact
     1801 ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag
     1861 gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc
     1921 aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc
     1981 agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt
     2041 tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gaatttcgta
     2101 tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt
     2161 tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc
     2221 agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc
     2281 ctaaagggtt tattgagaat atgtttttcg tttcagccaa tccctgggtg agtttcacca
     2341 gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca
     2401 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg
     2461 tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt
     2521 ggcagggcgg ggcgtaattt gatatcgagc tcgcttggac tcctgttgat agatccagta
     2581 atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt
     2641 attggt
//
