LOCUS       sequence-292553-        5677 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast 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 5677)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5677)
  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-m4046551
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        44..457
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1386..1403
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1404..1427
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1472..1490)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1841..2644
                     /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      2727..3709
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3787..3816
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3838..4425
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4437..4464)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4488..4679)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4680..5471,5472..5540))
                     /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     5303..5322
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5541..5643)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5644..5675)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 ctgctatgga ggtcaggtat gatttaaatg gtcagttaga gcggccgccc cacacaccat
       61 agcttcaaaa tgtttctact ccttttttac tcttccagat tttctcggac tccgcgcatc
      121 gccgtaccac ttcaaaacac ccaagcacag catactaaat ttcccctctt tcttcctcta
      181 gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa aaaagagacc gcctcgtttc
      241 tttttcttcg tcgaaaaagg caataaaaat ttttatcacg tttctttttc ttgaaaattt
      301 ttttttttga tttttttctc tttcgatgac ctcccattga tatttaagtt aataaacggt
      361 cttcaatttc tcaagtttca gtttcatttt tcttgttcta ttacaacttt ttttacttct
      421 tgctcattag aaagaaagca tagcaatcta atctaagttt taattacaaa atgggtttta
      481 gaaaaatggc attcccatct ggtaaagttg agggttgtat ggtacaagta acttgtggta
      541 caactacact taacggtctt tggcttgatg acgtagttta ctgtccaaga catgtgatct
      601 gcacctctga agacatgctt aaccctaatt atgaagattt actcattcgt aagtctaatc
      661 ataatttctt ggtacaggct ggtaatgttc aactcagggt tattggacat tctatgcaaa
      721 attgtgtact taagcttaag gttgatacag ccaatcctaa gacacctaag tataagtttg
      781 ttcgcattca accaggacag actttttcag tgttagcttg ttacaatggt tcaccatctg
      841 gtgtttacca atgtgctatg aggcccaatt tcactattaa gggttcattc cttaatggtt
      901 catgtggtag tgttggtttt aacatagatt atgactgtgt ctctttttgt tacatgcacc
      961 atatggaatt accaactgga gttcatgctg gcacagactt agaaggtaac ttttatggac
     1021 cttttgttga caggcaaaca gcacaagcag ctggtacgga cacaactatt acagttaatg
     1081 ttttagcttg gttgtacgct gctgttataa atggagacag gtggtttctc aatcgattta
     1141 ccacaactct taatgacttt aaccttgtgg ctatgaagta caattatgaa cctctaacac
     1201 aagaccatgt tgacatacta ggacctcttt ctgctcaaac tggaattgcc gttttagata
     1261 tgtgtgcttc attaaaagaa ttactgcaaa atggtatgaa tggacgtacc atattgggta
     1321 gtgctttatt agaagatgaa tttacacctt ttgatgttgt tagacaatgc tcaggtgtta
     1381 ctttcctcgt gccgagaggt tcttggagcc atccgcagtt tgaaaaataa acaggcccct
     1441 tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc cctcctccca
     1501 catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt
     1561 ttttttaata gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt
     1621 tctgtacaaa gcggccgccg aaccactagt gcactgcagt acacggtttc cttgaaattt
     1681 ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt
     1741 ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca
     1801 actgcacaga acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag ctacatataa
     1861 ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga
     1921 aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt
     1981 agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga
     2041 tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt
     2101 actcttcgag gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc
     2161 gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc
     2221 aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct
     2281 tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg
     2341 tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga
     2401 catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga
     2461 tgacaaggga gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg tttctacagg
     2521 atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga
     2581 gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa
     2641 ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat
     2701 ttaattatat cagttattac ccgagtaacg aagcatctgt gcttcatttt gtagaacaaa
     2761 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     2821 agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa
     2881 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     2941 agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt
     3001 gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt
     3061 ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg
     3121 ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact
     3181 ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg
     3241 catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat
     3301 agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta
     3361 tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag
     3421 ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga
     3481 ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata
     3541 tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg
     3601 caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcatc
     3661 agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctagc tagagaatag
     3721 gaacttcccg agcggccgcg tgttacaagc ttgactccag cgtaactgga ctgcaatcaa
     3781 ctcactggct caccttcacg ggtgggcctt tcttcggtag aaaatcaaag gatcttcttg
     3841 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3901 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaggtaac tggcttcagc
     3961 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4021 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4081 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4141 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4201 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4261 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4321 ccaggggtaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4381 catcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgca
     4441 gaaaggccca cccgaaggtg agccaggtga ttacatttgg gccctcatca gaggttttca
     4501 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     4561 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     4621 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     4681 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4741 gttgcctgac tccccgtcgt gtagataact acgatgcggg agggcttacc atctggcccc
     4801 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4861 cagccagccg ggagggccga gcgcagaagt gatcctgcaa ctttatccgc ctccatccag
     4921 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4981 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5041 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5101 gttagctcct tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt gttatcactc
     5161 atggttatgg cagcactgca taattctctt actgtcatgc catccgtgag atgcttttct
     5221 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5281 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5341 atcattggaa aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct gttgagatcc
     5401 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5461 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5521 cggaaatgtt gaatactcat tttagcttcc ttagctcctg aaaatctcga taactcaaaa
     5581 aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga
     5641 tcaagtcaaa agcctccggt cggaggcttt tgacttt
//
