LOCUS       sequence-292580-        2648 bp    DNA     circular SYN 14-APR-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 2648)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 2648)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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..2648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      208..242
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     primer_bind     270..289
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        270..288
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    289..313
                     /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             328..350
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1273..1293
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1294..1311
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      1432..2020
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1921..1940
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2126..2648,1..137)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2192..2211)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg
       61 tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt
      121 ggcagggcgg ggcgtaattt gatatcgagc tcgcttggac tcctgttgat agatccagta
      181 atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt
      241 attggtcatc gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc
      301 ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga ctcttctatg
      361 ggttttagaa aaatggcatt cccatctggt aaagttgagg gttgtatggt acaagtaact
      421 tgtggtacaa ctacacttaa cggtctttgg cttgatgacg tagtttactg tccaagacat
      481 gtgatctgca cctctgaaga catgcttaac cctaattatg aagatttact cattcgtaag
      541 tctaatcata atttcttggt acaggctggt aatgttcaac tcagggttat tggacattct
      601 atgcaaaatt gtgtacttaa gcttaaggtt gatacagcca atcctaagac acctaagtat
      661 aagtttgttc gcattcaacc aggacagact ttttcagtgt tagcttgtta caatggttca
      721 ccatctggtg tttaccaatg tgctatgagg cccaatttca ctattaaggg ttcattcctt
      781 aatggttcat gtggtagtgt tggttttaac atagattatg actgtgtctc tttttgttac
      841 atgcaccata tggaattacc aactggagtt catgctggca cagacttaga aggtaacttt
      901 tatggacctt ttgttgacag gcaaacagca caagcagctg gtacggacac aactattaca
      961 gttaatgttt tagcttggtt gtacgctgct gttataaatg gagacaggtg gtttctcaat
     1021 cgatttacca caactcttaa tgactttaac cttgtggcta tgaagtacaa ttatgaacct
     1081 ctaacacaag accatgttga catactagga cctctttctg ctcaaactgg aattgccgtt
     1141 ttagatatgt gtgcttcatt aaaagaatta ctgcaaaatg gtatgaatgg acgtaccata
     1201 ttgggtagtg ctttattaga agatgaattt acaccttttg atgttgttag acaatgctca
     1261 ggtgttactt tcgagaacct gtactttcaa agccaccatc atcaccatca ttaaccggct
     1321 tatcggtcag tttcacctga tttacgtaaa aacccgcttc ggcgggtttt tgcttttgga
     1381 ggggcagaaa gatgaatgac tgtccacgac gctataccca aaagaaaaag cttgagatcc
     1441 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     1501 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     1561 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     1621 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     1681 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     1741 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     1801 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     1861 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     1921 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     1981 atttttgtga tgctcgtcag gggggcggag cctatggaaa ttgatcgggc acgtaagagg
     2041 ttccaacttt caccataatg aaataagatc actaccgggc gtattttttg agttatcgag
     2101 attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga
     2161 tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac
     2221 ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa
     2281 gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga
     2341 atttcgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta
     2401 caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga
     2461 tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc
     2521 ctatttccct aaagggttta ttgagaatat gtttttcgtt tcagccaatc cctgggtgag
     2581 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac
     2641 catgggca
//
