LOCUS       sequence-300478-        6924 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 6924)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6924)
  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-m4046537
FEATURES             Location/Qualifiers
     source          1..6924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(253..1056)
                     /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"
     primer_bind     1407..1425
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(2568..2585)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        complement(3638..4079)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(3644..3667)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    3962..4079
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     terminator      4146..4177
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        4178..4280
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(4281..4349,4350..5141)
                     /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     complement(4499..4518)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             5142..5333
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      5357..5384
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(5396..5983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6005..6034)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      join(6112..6924,1..170)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 cgcgttgcat ttttgtttta caaaaatgaa gcacagattc ttcgttggta aaatagcgct
       61 ttcgcgttgc atttctgttc tgtaaaaatg cagctcagat tctttgtttg aaaaattagc
      121 gctctcgcgt tgcatttttg ttctacaaaa tgaagcacag atgcttcgtt actcgggtaa
      181 taactgatat aattaaattg aagctctaat ttgtgagttt agtatacatg catttactta
      241 taatacagtt ttttagtttt gctggccgca tcttctcaaa tatgcttccc agcctgcttt
      301 tctgtaacgt tcaccctcta ccttagcatc ccttcccttt gcaaatagtc ctcttccaac
      361 aataataatg tcagatcctg tagaaaccac atcatccacg gttctatact gttgacccaa
      421 tgcatctccc ttgtcatcta aacccacacc gggtgtcata atcaaccaat cgtaaccttc
      481 atctcttcca cccatgtctc tttgagcaat aaagccgata acaaaatctt tgtcgctctt
      541 cgcaatgtca acagtaccct tagtatattc tccagtagat agggagccct tgcatgacaa
      601 ttctgctaac atcaaaaggc ctctaggttc ctttgttact tcttctgccg cctgcttcaa
      661 accgctaaca atacctgggc ccaccacacc gtgtgcattc gtaatgtctg cccattctgc
      721 tattctgtat acacccgcag agtactgcaa tttgactgta ttaccaatgt cagcaaattt
      781 tctgtcctcg aagagtaaaa aattgtactt ggcggataat gcctttagcg gcttaactgt
      841 gccctccatg gaaaaatcag tcaagatatc cacatgtgtt tttagtaaac aaattttggg
      901 acctaatgct tcaactaact ccagtaattc cttggtggta cgaacatcca atgaagcaca
      961 caagtttgtt tgcttttcgt gcatgatatt aaatagcttg gcagcaacag gactaggatg
     1021 agtagcagca cgttccttat atgtagcttt cgacatgatt tatcttcgtt tcctgcacgt
     1081 ttttgttctg tgcagttggg ttaagaatac tgggcaattt catgtttctt caacactaca
     1141 tatgcgtata tataccaatc taagtctgtg ctccttcctt cgttcttcct tctgttcgga
     1201 gattaccgaa tcaaaaaaat ttcaaggaaa ccgtgtactg cagtgcacta gtggttcggc
     1261 ggccgctttg tacagaaaaa aaagaaaaat ttgaaatata aataacgttc ttaatactaa
     1321 cataactatt aaaaaaaata aatagggacc tagacttcag gttgtctaac tccttccttt
     1381 tcggttagag cggatgtggg aggagggcgt gaatgtaagc gtgacataac taattacatg
     1441 atatcgacaa aggaaaaggg gcctgtttac gtttgagcat cttttaatgc ttcgtcgacg
     1501 gtttgtctac cggaagcagc attaatcacg gcagtcctaa ctgcgtacca gaaggctgac
     1561 atttgtggga tgtttggcat aatttcaccc ttttgggcgt tctccatggt agcggcaatt
     1621 ctgggatcct tagccaattc ttcttcgtaa gatttcaaag cgacagcacc caatggcttg
     1681 tccttgttaa cggcctccaa accttcgtca gttaacaagt aattttccaa aaattcttta
     1741 gctagctcct tgtttggaga agcagcattg atgccagcgg atagaacacc gacaaagggc
     1801 ttggatggtt ggcctttgaa tgtaggcaaa acagtgacac catagtttac tttagaagta
     1861 tcaatgttac tccaggccca tggaccattt atggtcatag cagtttcacc cttgttgaaa
     1921 gcggcttcgg cgatagagta gtcagtgtca gcgttcatgt gtttattctt tattaaatca
     1981 accaagaagg tcaaaccagc ctttgcacca gcattatcaa cgccgacatc cttaatgtca
     2041 tacttaccgt tttcgtattt aaatgcgtaa ccaccgtcag cagcgattaa tggccaggtg
     2101 aaataaggtt cttgtaagtt aaacatcaaa gcgctttttc ccttggcctt tagctcttta
     2161 tccaaagctg gaatttcttc ccatgtcttt ggtgggttag gtaacaagtc cttattgtag
     2221 atcaacgata aggcttcaac agcaattgga tatgctataa gcttaccgtt gtatctgaca
     2281 gcatcccaag tgaatgggta caatttatct tggaaggcct tgtctggggt gatctcagcc
     2341 agcaaacctg actgagcata tccaccaaaa cgatcgtggg cccagaaaat aatgtcggga
     2401 ccatcaccag tagcagcaac ttgagggaat ttttcttcta acttgtctgg atgttcaacc
     2461 gtcaccttga tacctgtgtc cttttcaaat ttcttgccta cttctgccaa accgttgtaa
     2521 cccttatctc catttatcca aatgactagc ttaccttcct cgattttaga acctctcggc
     2581 acgagttgca atttcgggta gaaggtttca acatgaccgt ctttgcacca caacataaat
     2641 gagatttccg tatagtcaat ggttaccttg acaaccttgg aaacaactga caagtcttga
     2701 gacttaatta tttctacgaa atcatctagt aacaagtcaa tgacggaaca aacacactta
     2761 ctcgaaccgg tttgagcatc agtgatgaag tagtttttaa cggtagagtc catagggata
     2821 aaatcttcca actcgaatgg tgattccttg aatcttttag ccagtccaat tagcaagtgt
     2881 aaaccaccca gctgggaatg agagaagtca ccgtagacaa tgtgttcaaa agcataacct
     2941 tccaacttgt aacgttcgat gaactcgtcc atagctagtt caagaaaatc gatttccatt
     3001 tgagatcgtg gtttaaattc ttgcaaattg cgagactgag tgaagtaagt ttctggtaat
     3061 tgttgcacaa caccatcgac cttcttatag taattgaatt gagtcttaac ggcttcgcca
     3121 atcaacgtga cgccgttcaa ggacgcttgc tttggaccta cagaaggttg tagaccttta
     3181 acactacctt ctgtaatcaa aacaccgttc cttgcgtttc tgaagaggtc gacttgacca
     3241 tctactctac cgtcaaagaa aacggttaat ggagcacaga tagtctcggt tggcttttta
     3301 gcaatgtcag tcatcgagca gacacctata gttgagatat gggccggagc atctctcttg
     3361 taatcccaaa tgacggtatt agcagcgatg tcaacaccca aattgttcaa aatcttaact
     3421 tctgggactg gtttgatgtt acgcttcgcc caaagttcaa atgcaacgtt gacgggtaat
     3481 gtcgtcttat tttcgaacaa ttccacatca acgccgtcga ccttagtata aacggtgtta
     3541 tttataatgg agacaggaac ctcaccctgt tgcccatcaa agtgtccttt gttcactaca
     3601 ttgaaggcaa cgttttccag gctcattata gttttttctc cttgacgtta aagtatagag
     3661 gtatattaac aattttttgt tgatactttt atgacatttg aataagaagt aatacaaact
     3721 gaaaatgttg aaagtattag ttaaagtggt tatgcagctt ttccatttat atatctgtta
     3781 atagatcaaa aatcatcgct tcgctgatta attaccccag aaataaggct aaaaaactaa
     3841 tcgcattatc atcctatggt tgttaatttg attcgttaat ttgaaggttt gtggggccag
     3901 gttactgcca atttttcctc ttcataacca taaaagctag tattgtagaa tctttattgt
     3961 tcggagcagt gcggcgcgag gcacatctgc gtttcaggaa cgcgaccggt gaagacgagg
     4021 acgcacggag gagagtcttc cgtcggaggg ctgtcgcccg ctcggcggct tctaatccgt
     4081 acttcaatat agcaagcggc cgctctaact gaccatttaa atcatacctg acctccatag
     4141 cagaaagtca aaagcctccg accggaggct tttgacttga tcggcacgta agaggttcca
     4201 actttcacca taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt
     4261 caggagctaa ggaagctaaa atgagtattc aacatttccg tgtcgccctt attccctttt
     4321 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4381 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4441 tccttgagag tttacgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4501 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4561 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat ctcacggatg
     4621 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4681 acttacttct ggcaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4741 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4801 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     4861 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4921 ttgcaggatc acttctgcgc tcggccctcc cggctggctg gtttattgct gataaatctg
     4981 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5041 cccgcatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5101 agatcgctga gataggtgcc tcactgatta agcattggta agtgaccaaa caggaaaaaa
     5161 ccgcccttaa catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca
     5221 acgagctgga cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg
     5281 agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgatgagggc
     5341 ccaaatgtaa tcacctggct caccttcggg tgggcctttc tgcgttgctg gcgtttttcc
     5401 ataggctccg cccccctgac gagcatcaca aaaatcgatg ctcaagtcag aggtggcgaa
     5461 acccgacagg actataaaga taccaggcgt ttacccctgg aagctccctc gtgcgctctc
     5521 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5581 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5641 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5701 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5761 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5821 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttacctcgg
     5881 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5941 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatttt
     6001 ctaccgaaga aaggcccacc cgtgaaggtg agccagtgag ttgattgcag tccagttacg
     6061 ctggagtcaa gcttgtaaca cgcggccgct cgggaagttc ctattctcta gctagaaagt
     6121 ataggaactt cagagcgctt ttgaaaacca aaagcgctct gatgacgcac tttcaaaaaa
     6181 ccaaaaacgc accggactgt aacgagctac taaaatattg cgaataccgc ttccacaaac
     6241 attgctcaaa agtatctctt tgctatatat ctctgtgcta tatccctata taacctaccc
     6301 atccaccttt cgctccttga acttgcatct aaactcgacc tctacatttt ttatgtttat
     6361 ctctagtatt actctttaga caaaaaaatt gtagtaagaa ctattcatag agtgaatcga
     6421 aaacaatacg aaaatgtaaa catttcctat acgtagtata tagagacaaa atagaagaaa
     6481 ccgttcataa ttttctgacc aatgaagaat catcaacgct atcactttct gttcacaaag
     6541 tatgcgcaat ccacatcggt atagaatata atcggggatg cctttatctt gaaaaaatgc
     6601 acccgcagct tcgctagtaa tcagtaaacg cgggaagtgg agtcaggctt tttttatgga
     6661 agagaaaata gacaccaaag tagccttctt ctaaccttaa cggacctaca gtgcaaaaag
     6721 ttatcaagag actgcattat agagcgcaca aaggagaaaa aaagtaatct aagatgcttt
     6781 gttagaaaaa tagcgctctc gggatgcatt tttgtagaac aaaaaagaag tatagattct
     6841 ttgttggtaa aatagcgctc tcgcgttgca tttctgttct gtaaaaatgc agctcagatt
     6901 ctttgtttga aaaattagcg ctct
//
