LOCUS       sequence-298027-        5616 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 3C-like proteinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5616)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5616)
  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-m4046440
FEATURES             Location/Qualifiers
     source          1..5616
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1291..1308
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1309..1326
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1371..1389)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1740..2543
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      2626..3608
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3686..3715
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3737..4324
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4336..4363)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4387..4578)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4579..5370,5371..5439))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5202..5221
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5440..5542)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5543..5574)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcttgaa aatttttttt tttgattttt ttctctttcg atgacctccc
      241 attgatattt aagttaataa acggtcttca atttctcaag tttcagtttc atttttcttg
      301 ttctattaca acttttttta cttcttgctc attagaaaga aagcatagca atctaatcta
      361 agttttaatt acaaaatggg ttttagaaaa atggcattcc catctggtaa agttgagggt
      421 tgtatggtac aagtaacttg tggtacaact acacttaacg gtctttggct tgatgacgta
      481 gtttactgtc caagacatgt gatctgcacc tctgaagaca tgcttaaccc taattatgaa
      541 gatttactca ttcgtaagtc taatcataat ttcttggtac aggctggtaa tgttcaactc
      601 agggttattg gacattctat gcaaaattgt gtacttaagc ttaaggttga tacagccaat
      661 cctaagacac ctaagtataa gtttgttcgc attcaaccag gacagacttt ttcagtgtta
      721 gcttgttaca atggttcacc atctggtgtt taccaatgtg ctatgaggcc caatttcact
      781 attaagggtt cattccttaa tggttcatgt ggtagtgttg gttttaacat agattatgac
      841 tgtgtctctt tttgttacat gcaccatatg gaattaccaa ctggagttca tgctggcaca
      901 gacttagaag gtaactttta tggacctttt gttgacaggc aaacagcaca agcagctggt
      961 acggacacaa ctattacagt taatgtttta gcttggttgt acgctgctgt tataaatgga
     1021 gacaggtggt ttctcaatcg atttaccaca actcttaatg actttaacct tgtggctatg
     1081 aagtacaatt atgaacctct aacacaagac catgttgaca tactaggacc tctttctgct
     1141 caaactggaa ttgccgtttt agatatgtgt gcttcattaa aagaattact gcaaaatggt
     1201 atgaatggac gtaccatatt gggtagtgct ttattagaag atgaatttac accttttgat
     1261 gttgttagac aatgctcagg tgttactttc ctcgtgccga gaggttctca ccatcatcac
     1321 catcattaaa caggcccctt ttcctttgtc gatatcatgt aattagttat gtcacgctta
     1381 cattcacgcc ctcctcccac atccgctcta accgaaaagg aaggagttag acaacctgaa
     1441 gtctaggtcc ctatttattt tttttaatag ttatgttagt attaagaacg ttatttatat
     1501 ttcaaatttt tctttttttt ctgtacaaag cggccgccga accactagtg cactgcagta
     1561 cacggtttcc ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga
     1621 aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt
     1681 gcccagtatt cttaacccaa ctgcacagaa caaaaacgtg caggaaacga agataaatca
     1741 tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc
     1801 tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca
     1861 ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac
     1921 atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat
     1981 ccgccaagta caatttttta ctcttcgagg acagaaaatt tgctgacatt ggtaatacag
     2041 tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg
     2101 cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa
     2161 caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta
     2221 ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg
     2281 gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga
     2341 cacccggtgt gggtttagat gacaagggag atgcattggg tcaacagtat agaaccgtgg
     2401 atgatgtggt ttctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg
     2461 gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga
     2521 gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact
     2581 cacaaattag agcttcaatt taattatatc agttattacc cgagtaacga agcatctgtg
     2641 cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     2701 agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca acgaagaatc
     2761 tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt caaacaaaga
     2821 atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt taccaacaaa
     2881 gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat ttttctaaca
     2941 aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc tcttgataac
     3001 tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct attttctctt
     3061 ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa gctgcgggtg
     3121 cattttttca agataaaggc atccccgatt atattctata ccgatgtgga ttgcgcatac
     3181 tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat tatgaacggt
     3241 ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc gtattgtttt
     3301 cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta atactagaga
     3361 taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga aaggtggatg
     3421 ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt ttgagcaatg
     3481 tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg cgtttttggt
     3541 tttttgaaag tgcgtcatca gagcgctttt ggttttcaaa agcgctctga agttcctata
     3601 ctttctagct agagaatagg aacttcccga gcggccgcgt gttacaagct tgactccagc
     3661 gtaactggac tgcaatcaac tcactggctc accttcacgg gtgggccttt cttcggtaga
     3721 aaatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3781 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3841 cgaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3901 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3961 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4021 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4081 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4141 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4201 agagcgcacg agggagcttc caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt
     4261 tcgccacctc tgacttgagc atcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4321 gaaaaacgcc agcaacgcag aaaggcccac ccgaaggtga gccaggtgat tacatttggg
     4381 ccctcatcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     4441 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     4501 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     4561 tttttcctgt ttggtcactt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4621 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatgcggga
     4681 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4741 agatttatca gcaataaacc agccagccgg gagggccgag cgcagaagtg atcctgcaac
     4801 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4861 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4921 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4981 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgcca gaagtaagtt
     5041 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5101 atccgtgaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5161 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5221 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgtaaac tctcaaggat
     5281 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5341 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5401 aaagggaata agggcgacac ggaaatgttg aatactcatt ttagcttcct tagctcctga
     5461 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
     5521 ggaacctctt acgtgccgat caagtcaaaa gcctccggtc ggaggctttt gactttctgc
     5581 tatggaggtc aggtatgatt taaatggtca gttaga
//
