LOCUS       sequence-287825-        2652 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 2652)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2652)
  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-m4046781
FEATURES             Location/Qualifiers
     source          1..2652
                     /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..1073
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     rep_origin      1190..1778
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1679..1698
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1884..2543
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1950..1969)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2614..2648
                     /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 agcgagcaga agcttatttc cgaagaagat ctctaaccgg
     1081 cttatcggtc agtttcacct gatttacgta aaaacccgct tcggcgggtt tttgcttttg
     1141 gaggggcaga aagatgaatg actgtccacg acgctatacc caaaagaaat tgagatcctt
     1201 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     1261 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     1321 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     1381 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     1441 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     1501 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     1561 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     1621 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     1681 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     1741 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaatt gatcgggcac gtaagaggtt
     1801 ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat
     1861 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
     1921 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
     1981 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     2041 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     2101 ttcgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
     2161 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
     2221 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
     2281 atttccctaa agggtttatt gagaatatgt ttttcgtttc agccaatccc tgggtgagtt
     2341 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     2401 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     2461 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
     2521 atgagtggca gggcggggcg taatttgata tcgagctcgc ttggactcct gttgatagat
     2581 ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg
     2641 ttttttattg gt
//
