LOCUS       sequence-287575-        5845 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 endoRNAse.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5845)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5845)
  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-m4046471
FEATURES             Location/Qualifiers
     source          1..5845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1472..1489
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1490..1555
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1600..1618)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1969..2772
                     /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      2855..3837
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3915..3944
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3966..4553
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4565..4592)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4616..4807)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4808..5599,5600..5668))
                     /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     5431..5450
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5669..5771)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5772..5803)
                     /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 gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atgagcctgg aaaacgttgc cttcaatgta gtgaacaaag gacactttga
      481 tgggcaacag ggtgaggttc ctgtctccat tataaataac accgtttata ctaaggtcga
      541 cggcgttgat gtggaattgt tcgaaaataa gacgacatta cccgtcaacg ttgcatttga
      601 actttgggcg aagcgtaaca tcaaaccagt cccagaagtt aagattttga acaatttggg
      661 tgttgacatc gctgctaata ccgtcatttg ggattacaag agagatgctc cggcccatat
      721 ctcaactata ggtgtctgct cgatgactga cattgctaaa aagccaaccg agactatctg
      781 tgctccatta accgttttct ttgacggtag agtagatggt caagtcgacc tcttcagaaa
      841 cgcaaggaac ggtgttttga ttacagaagg tagtgttaaa ggtctacaac cttctgtagg
      901 tccaaagcaa gcgtccttga acggcgtcac gttgattggc gaagccgtta agactcaatt
      961 caattactat aagaaggtcg atggtgttgt gcaacaatta ccagaaactt acttcactca
     1021 gtctcgcaat ttgcaagaat ttaaaccacg atctcaaatg gaaatcgatt ttcttgaact
     1081 agctatggac gagttcatcg aacgttacaa gttggaaggt tatgcttttg aacacattgt
     1141 ctacggtgac ttctctcatt cccagctggg tggtttacac ttgctaattg gactggctaa
     1201 aagattcaag gaatcaccat tcgagttgga agattttatc cctatggact ctaccgttaa
     1261 aaactacttc atcactgatg ctcaaaccgg ttcgagtaag tgtgtttgtt ccgtcattga
     1321 cttgttacta gatgatttcg tagaaataat taagtctcaa gacttgtcag ttgtttccaa
     1381 ggttgtcaag gtaaccattg actatacgga aatctcattt atgttgtggt gcaaagacgg
     1441 tcatgttgaa accttctacc cgaaattgca actcgtgccg agaggttctg attataaaga
     1501 tcatgatggt gattataaag atcatgatat tgattataaa gatgatgatg ataaataaac
     1561 aggccccttt tcctttgtcg atatcatgta attagttatg tcacgcttac attcacgccc
     1621 tcctcccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc
     1681 tatttatttt ttttaatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt
     1741 cttttttttc tgtacaaagc ggccgccgaa ccactagtgc actgcagtac acggtttcct
     1801 tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga
     1861 cttagattgg tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc
     1921 ttaacccaac tgcacagaac aaaaacgtgc aggaaacgaa gataaatcat gtcgaaagct
     1981 acatataagg aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc
     2041 atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta
     2101 ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc
     2161 ttgactgatt tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac
     2221 aattttttac tcttcgagga cagaaaattt gctgacattg gtaatacagt caaattgcag
     2281 tactctgcgg gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg
     2341 gtgggcccag gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct
     2401 agaggccttt tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat
     2461 actaagggta ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct
     2521 caaagagaca tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg
     2581 ggtttagatg acaagggaga tgcattgggt caacagtata gaaccgtgga tgatgtggtt
     2641 tctacaggat ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct
     2701 aaggtagagg gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc
     2761 cagcaaaact aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga
     2821 gcttcaattt aattatatca gttattaccc gagtaacgaa gcatctgtgc ttcattttgt
     2881 agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     2941 tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt
     3001 tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     3061 atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact
     3121 tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag
     3181 attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg
     3241 taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa
     3301 gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa
     3361 gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag
     3421 aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt
     3481 tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct
     3541 atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa
     3601 aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata
     3661 tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc
     3721 ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt ttttgaaagt
     3781 gcgtcatcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac tttctagcta
     3841 gagaatagga acttcccgag cggccgcgtg ttacaagctt gactccagcg taactggact
     3901 gcaatcaact cactggctca ccttcacggg tgggcctttc ttcggtagaa aatcaaagga
     3961 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4021 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaggtaactg
     4081 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4141 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4201 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4261 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4321 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4381 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4441 gggagcttcc aggggtaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4501 gacttgagca tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4561 gcaacgcaga aaggcccacc cgaaggtgag ccaggtgatt acatttgggc cctcatcaga
     4621 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     4681 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     4741 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     4801 tggtcactta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4861 catccatagt tgcctgactc cccgtcgtgt agataactac gatgcgggag ggcttaccat
     4921 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4981 caataaacca gccagccggg agggccgagc gcagaagtga tcctgcaact ttatccgcct
     5041 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5101 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5161 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5221 aaaaagcggt tagctccttc ggtcctccga tcgttgccag aagtaagttg gccgcagtgt
     5281 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtgagat
     5341 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5401 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5461 aagtgctcat cattggaaaa cgttcttcgg ggcgtaaact ctcaaggatc ttaccgctgt
     5521 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5581 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5641 gggcgacacg gaaatgttga atactcattt tagcttcctt agctcctgaa aatctcgata
     5701 actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta
     5761 cgtgccgatc aagtcaaaag cctccggtcg gaggcttttg actttctgct atggaggtca
     5821 ggtatgattt aaatggtcag ttaga
//
