LOCUS       sequence-300253-        2654 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 2654)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2654)
  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-m4046895
FEATURES             Location/Qualifiers
     source          1..2654
                     /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="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     rep_origin      1188..1776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1677..1696
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             1882..2541
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1948..1967)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2612..2646
                     /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 agctggagcc atccgcagtt tgaaaaataa ccggcttatc
     1081 ggtcagtttc acctgattta cgtaaaaacc cgcttcggcg ggtttttgct tttggagggg
     1141 cagaaagatg aatgactgtc cacgacgcta tacccaaaag aaaaagcttg agatcctttt
     1201 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     1261 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     1321 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     1381 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     1441 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     1501 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     1561 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     1621 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     1681 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     1741 ttgtgatgct cgtcaggggg gcggagccta tggaaattga tcgggcacgt aagaggttcc
     1801 aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt
     1861 tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata
     1921 tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat
     1981 aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac
     2041 aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattt
     2101 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
     2161 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
     2221 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
     2281 ttccctaaag ggtttattga gaatatgttt ttcgtttcag ccaatccctg ggtgagtttc
     2341 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
     2401 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
     2461 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
     2521 gagtggcagg gcggggcgta atttgatatc gagctcgctt ggactcctgt tgatagatcc
     2581 agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt
     2641 ttttattggt catc
//
