LOCUS       sequence-287801-        2649 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 2649)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2649)
  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-m4046771
FEATURES             Location/Qualifiers
     source          1..2649
                     /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..1070
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      1187..1775
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1676..1695
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1881..2540
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1947..1966)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2611..2645
                     /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 agctacccgt atgacgttcc tgattatgcc taaccggctt
     1081 atcggtcagt ttcacctgat ttacgtaaaa acccgcttcg gcgggttttt gcttttggag
     1141 gggcagaaag atgaatgact gtccacgacg ctatacccaa aagaaattga gatccttttt
     1201 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1261 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1321 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1381 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1441 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1501 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1561 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     1621 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1681 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     1741 tgtgatgctc gtcagggggg cggagcctat ggaaattgat cgggcacgta agaggttcca
     1801 actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt
     1861 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat
     1921 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata
     1981 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca
     2041 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaatttc
     2101 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg
     2161 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc
     2221 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt
     2281 tccctaaagg gtttattgag aatatgtttt tcgtttcagc caatccctgg gtgagtttca
     2341 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg
     2401 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg
     2461 ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg
     2521 agtggcaggg cggggcgtaa tttgatatcg agctcgcttg gactcctgtt gatagatcca
     2581 gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt
     2641 tttattggt
//
