LOCUS       sequence-291438-        6877 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 6877)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6877)
  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-m4046523
FEATURES             Location/Qualifiers
     source          1..6877
                     /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"
     primer_bind     complement(2632..2650)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3001..3804
                     /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      3887..4869
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4947..4976
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4998..5585
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5597..5624)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5648..5839)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5840..6631,6632..6700))
                     /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     6463..6482
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6701..6803)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6804..6835)
                     /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 agaggttcta aaatcgagga
     1501 aggtaagcta gtcatttgga taaatggaga taagggttac aacggtttgg cagaagtagg
     1561 caagaaattt gaaaaggaca caggtatcaa ggtgacggtt gaacatccag acaagttaga
     1621 agaaaaattc cctcaagttg ctgctactgg tgatggtccc gacattattt tctgggccca
     1681 cgatcgtttt ggtggatatg ctcagtcagg tttgctggct gagatcaccc cagacaaggc
     1741 cttccaagat aaattgtacc cattcacttg ggatgctgtc agatacaacg gtaagcttat
     1801 agcatatcca attgctgttg aagccttatc gttgatctac aataaggact tgttacctaa
     1861 cccaccaaag acatgggaag aaattccagc tttggataaa gagctaaagg ccaagggaaa
     1921 aagcgctttg atgtttaact tacaagaacc ttatttcacc tggccattaa tcgctgctga
     1981 cggtggttac gcatttaaat acgaaaacgg taagtatgac attaaggatg tcggcgttga
     2041 taatgctggt gcaaaggctg gtttgacctt cttggttgat ttaataaaga ataaacacat
     2101 gaacgctgac actgactact ctatcgccga agccgctttc aacaagggtg aaactgctat
     2161 gaccataaat ggtccatggg cctggagtaa cattgatact tctaaagtaa actatggtgt
     2221 cactgttttg cctacattca aaggccaacc atccaagccc tttgtcggtg ttctatccgc
     2281 tggcatcaat gctgcttctc caaacaagga gctagctaaa gaatttttgg aaaattactt
     2341 gttaactgac gaaggtttgg aggccgttaa caaggacaag ccattgggtg ctgtcgcttt
     2401 gaaatcttac gaagaagaat tggctaagga tcccagaatt gccgctacca tggagaacgc
     2461 ccaaaagggt gaaattatgc caaacatccc acaaatgtca gccttctggt acgcagttag
     2521 gactgccgtg attaatgctg cttccggtag acaaaccgtc gacgaagcat taaaagatgc
     2581 tcaaacgtaa acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt
     2641 acattcacgc cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga
     2701 agtctaggtc cctatttatt ttttttaata gttatgttag tattaagaac gttatttata
     2761 tttcaaattt ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt
     2821 acacggtttc cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg
     2881 aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat
     2941 tgcccagtat tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc
     3001 atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag
     3061 ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc
     3121 accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca
     3181 catgtggata tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta
     3241 tccgccaagt acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca
     3301 gtcaaattgc agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat
     3361 gcacacggtg tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta
     3421 acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct
     3481 actggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc
     3541 ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg
     3601 acacccggtg tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg
     3661 gatgatgtgg tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag
     3721 ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg
     3781 agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac
     3841 tcacaaatta gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt
     3901 gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     3961 gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat
     4021 ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     4081 aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa
     4141 agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac
     4201 aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa
     4261 ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct
     4321 tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt
     4381 gcattttttc aagataaagg catccccgat tatattctat accgatgtgg attgcgcata
     4441 ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg
     4501 tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt
     4561 tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag
     4621 ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat
     4681 gggtaggtta tatagggata tagcacagag atatatagca aagagatact tttgagcaat
     4741 gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg
     4801 ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat
     4861 actttctagc tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag
     4921 cgtaactgga ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag
     4981 aaaatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5041 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5101 ccgaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5161 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5221 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5281 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5341 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5401 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5461 gagagcgcac gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt
     5521 ttcgccacct ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat
     5581 ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg
     5641 gccctcatca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     5701 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     5761 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     5821 ttttttcctg tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct
     5881 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg
     5941 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6001 cagatttatc agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa
     6061 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6121 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6181 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6241 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt
     6301 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     6361 catccgtgag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6421 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6481 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga
     6541 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6601 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6661 aaaagggaat aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg
     6721 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     6781 tggaacctct tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg
     6841 ctatggaggt caggtatgat ttaaatggtc agttaga
//
