LOCUS       sequence-297631-        6838 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 endoRNAse.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6838)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6838)
  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..6838
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        10..408
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1463..1480
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2623..2641)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2992..3795
                     /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      3878..4848
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4926..4955
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4977..5564
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5576..5603)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5627..5818)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5819..6607)
                     /codon_start=1
                     /gene="bla(M)"
                     /product="beta-lactamase lacking the signal sequence"
                     /label=bla(M)
                     /note="allows cytosolic expression of beta-lactamase"
                     /translation="PETLVKVKDAEDQLGARVGYIELDLNSGKILESLRPEERFPMMST
                     FKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMS
                     DNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATT
                     LRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSALPAGWFIADKSGAGERGSRGIIAA
                     LGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     primer_bind     6442..6461
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6662..6764)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6765..6796)
                     /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 ttttcttttt
      241 cttgaaaatt tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt
      301 taataaacgg tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt
      361 tttttacttc ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttaattacaa
      421 aatgagcctg gaaaacgttg ccttcaatgt agtgaacaaa ggacactttg atgggcaaca
      481 gggtgaggtt cctgtctcca ttataaataa caccgtttat actaaggtcg acggcgttga
      541 tgtggaattg ttcgaaaata agacgacatt acccgtcaac gttgcatttg aactttgggc
      601 gaagcgtaac atcaaaccag tcccagaagt taagattttg aacaatttgg gtgttgacat
      661 cgctgctaat accgtcattt gggattacaa gagagatgct ccggcccata tctcaactat
      721 aggtgtctgc tcgatgactg acattgctaa aaagccaacc gagactatct gtgctccatt
      781 aaccgttttc tttgacggta gagtagatgg tcaagtcgac ctcttcagaa acgcaaggaa
      841 cggtgttttg attacagaag gtagtgttaa aggtctacaa ccttctgtag gtccaaagca
      901 agcgtccttg aacggcgtca cgttgattgg cgaagccgtt aagactcaat tcaattacta
      961 taagaaggtc gatggtgttg tgcaacaatt accagaaact tacttcactc agtctcgcaa
     1021 tttgcaagaa tttaaaccac gatctcaaat ggaaatcgat tttcttgaac tagctatgga
     1081 cgagttcatc gaacgttaca agttggaagg ttatgctttt gaacacattg tctacggtga
     1141 cttctctcat tcccagctgg gtggtttaca cttgctaatt ggactggcta aaagattcaa
     1201 ggaatcacca ttcgagttgg aagattttat ccctatggac tctaccgtta aaaactactt
     1261 catcactgat gctcaaaccg gttcgagtaa gtgtgtttgt tccgtcattg acttgttact
     1321 agatgatttc gtagaaataa ttaagtctca agacttgtca gttgtttcca aggttgtcaa
     1381 ggtaaccatt gactatacgg aaatctcatt tatgttgtgg tgcaaagacg gtcatgttga
     1441 aaccttctac ccgaaattgc aactcgtgcc gagaggttct aaaatcgagg aaggtaagct
     1501 agtcatttgg ataaatggag ataagggtta caacggtttg gcagaagtag gcaagaaatt
     1561 tgaaaaggac acaggtatca aggtgacggt tgaacatcca gacaagttag aagaaaaatt
     1621 ccctcaagtt gctgctactg gtgatggtcc cgacattatt ttctgggccc acgatcgttt
     1681 tggtggatat gctcagtcag gtttgctggc tgagatcacc ccagacaagg ccttccaaga
     1741 taaattgtac ccattcactt gggatgctgt cagatacaac ggtaagctta tagcatatcc
     1801 aattgctgtt gaagccttat cgttgatcta caataaggac ttgttaccta acccaccaaa
     1861 gacatgggaa gaaattccag ctttggataa agagctaaag gccaagggaa aaagcgcttt
     1921 gatgtttaac ttacaagaac cttatttcac ctggccatta atcgctgctg acggtggtta
     1981 cgcatttaaa tacgaaaacg gtaagtatga cattaaggat gtcggcgttg ataatgctgg
     2041 tgcaaaggct ggtttgacct tcttggttga tttaataaag aataaacaca tgaacgctga
     2101 cactgactac tctatcgccg aagccgcttt caacaagggt gaaactgcta tgaccataaa
     2161 tggtccatgg gcctggagta acattgatac ttctaaagta aactatggtg tcactgtttt
     2221 gcctacattc aaaggccaac catccaagcc ctttgtcggt gttctatccg ctggcatcaa
     2281 tgctgcttct ccaaacaagg agctagctaa agaatttttg gaaaattact tgttaactga
     2341 cgaaggtttg gaggccgtta acaaggacaa gccattgggt gctgtcgctt tgaaatctta
     2401 cgaagaagaa ttggctaagg atcccagaat tgccgctacc atggagaacg cccaaaaggg
     2461 tgaaattatg ccaaacatcc cacaaatgtc agccttctgg tacgcagtta ggactgccgt
     2521 gattaatgct gcttccggta gacaaaccgt cgacgaagca ttaaaagatg ctcaaacgta
     2581 aacaggcccc ttttcctttg tcgatatcat gtaattagtt atgtcacgct tacattcacg
     2641 ccctcctccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt
     2701 ccctatttat tttttttaat agttatgtta gtattaagaa cgttatttat atttcaaatt
     2761 tttctttttt ttctgtacaa agcggccgcc gaaccactag tgcactgcag tacacggttt
     2821 ccttgaaatt tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac
     2881 agacttagat tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta
     2941 ttcttaaccc aactgcacag aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa
     3001 gctacatata aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat
     3061 atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa
     3121 ttactggagt tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat
     3181 atcttgactg atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag
     3241 tacaattttt tactcttcga ggacagaaaa tttgctgaca ttggtaatac agtcaaattg
     3301 cagtactctg cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt
     3361 gtggtgggcc caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa
     3421 cctagaggcc ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa
     3481 tatactaagg gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt
     3541 gctcaaagag acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt
     3601 gtgggtttag atgacaaggg agatgcattg ggtcaacagt atagaaccgt ggatgatgtg
     3661 gtttctacag gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat
     3721 gctaaggtag agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc
     3781 ggccagcaaa actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt
     3841 agagcttcaa tttaattata tcagttatta cccgagtaac gaagcatctg tgcttcattt
     3901 tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     3961 ttttacagaa cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc
     4021 atttttgtaa aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     4081 tgcattttta cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat
     4141 acttctaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt
     4201 tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt
     4261 taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc
     4321 cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc
     4381 atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata
     4441 gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat
     4501 atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt
     4561 tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag
     4621 gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat
     4681 agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc
     4741 aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca
     4801 gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg
     4861 aacttcccga gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac
     4921 tcactggctc accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga
     4981 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5041 gtggtttgtt tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca
     5101 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5161 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5221 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5281 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5341 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5401 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5461 caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5521 atcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag
     5581 aaaggcccac ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac
     5641 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     5701 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     5761 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt
     5821 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5881 ttgcctgact ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca
     5941 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6001 agccagccgg gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt
     6061 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6121 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6181 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6241 ttagctcctt cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca
     6301 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg
     6361 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6421 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6481 tcattggaaa acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca
     6541 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6601 tttctgggtg agcaaacgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6661 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc
     6721 ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg
     6781 tcggaggctt ttgactttct gctatggagg tcaggtatga tttaaatggt cagttaga
//
