LOCUS       sequence-298539-        5755 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 5755)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5755)
  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-m4046625
FEATURES             Location/Qualifiers
     source          1..5755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(1510..1528)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1879..2682
                     /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      2765..3747
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3825..3854
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3876..4463
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4475..4502)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4526..4717)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4718..5509,5510..5578))
                     /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     5341..5360
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5579..5681)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5682..5713)
                     /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 atgttagaaa atgtggcttt taatgttgta aataagggac actttgatgg
      481 acaacagggt gaagtaccag tttctatcat taataacact gtttacacaa aagttgatgg
      541 tgttgatgta gaattgtttg aaaataaaac aacattacct gttaatgtag catttgagct
      601 ttgggctaag cgcaacatta aaccagtacc agaggtgaaa atactcaata atttgggtgt
      661 ggacattgct gctaatactg tgatctggga ctacaaaaga gatgctccag cacatatatc
      721 tactattggt gtttgttcta tgactgacat agccaagaaa ccaactgaaa cgatttgtgc
      781 accactcact gtcttttttg atggtagagt tgatggtcaa gtagacttat ttagaaatgc
      841 ccgtaatggt gttcttatta cagaaggtag tgttaaaggt ttacaaccat ctgtaggtcc
      901 caaacaagct agtcttaatg gagtcacatt aattggagaa gccgtaaaaa cacagttcaa
      961 ttattataag aaagttgatg gtgttgtcca acaattacct gaaacttact ttactcagag
     1021 tagaaattta caagaattta aacccaggag tcaaatggaa attgatttct tagaattagc
     1081 tatggatgaa ttcattgaac ggtataaatt agaaggctat gccttcgaac atatcgttta
     1141 tggagatttt agtcatagtc agttaggtgg tttacatcta ctgattggac tagctaaacg
     1201 ttttaaggaa tcaccttttg aattagaaga ttttattcct atggacagta cagttaaaaa
     1261 ctatttcata acagatgcgc aaacaggttc atctaagtgt gtgtgttctg ttattgattt
     1321 attacttgat gattttgttg aaataataaa atcccaagat ttatctgtag tttctaaggt
     1381 tgtcaaagtg actattgact atacagaaat ttcatttatg ctttggtgta aagatggcca
     1441 tgtagaaaca ttttacccaa aattataaac aggccccttt tcctttgtcg atatcatgta
     1501 attagttatg tcacgcttac attcacgccc tcctcccaca tccgctctaa ccgaaaagga
     1561 aggagttaga caacctgaag tctaggtccc tatttatttt ttttaatagt tatgttagta
     1621 ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacaaagc ggccgccgaa
     1681 ccactagtgc actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa
     1741 cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt
     1801 gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc
     1861 aggaaacgaa gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct
     1921 agtcctgttg ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct
     1981 tcattggatg ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa
     2041 atttgtttac taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt
     2101 aagccgctaa aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt
     2161 gctgacattg gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa
     2221 tgggcagaca ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag
     2281 caggcggcag aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca
     2341 tgcaagggct ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc
     2401 gacaaagatt ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt
     2461 tacgattggt tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt
     2521 caacagtata gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga
     2581 agaggactat ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca
     2641 ggctgggaag catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta
     2701 aatgcatgta tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc
     2761 gagtaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat
     2821 ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct
     2881 attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg
     2941 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga
     3001 gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga
     3061 gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat
     3121 aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact
     3181 ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt
     3241 actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac
     3301 cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg
     3361 tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt
     3421 tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc
     3481 taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc
     3541 aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa
     3601 gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac
     3661 agtccggtgc gtttttggtt ttttgaaagt gcgtcatcag agcgcttttg gttttcaaaa
     3721 gcgctctgaa gttcctatac tttctagcta gagaatagga acttcccgag cggccgcgtg
     3781 ttacaagctt gactccagcg taactggact gcaatcaact cactggctca ccttcacggg
     3841 tgggcctttc ttcggtagaa aatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3901 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3961 gagctaccaa ctctttttcc gaggtaactg gcttcagcag agcgcagata ccaaatactg
     4021 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4081 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4141 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4201 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4261 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4321 gcggcagggt cggaacagga gagcgcacga gggagcttcc aggggtaaac gcctggtatc
     4381 tttatagtcc tgtcgggttt cgccacctct gacttgagca tcgatttttg tgatgctcgt
     4441 caggggggcg gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc cgaaggtgag
     4501 ccaggtgatt acatttgggc cctcatcaga ggttttcacc gtcatcaccg aaacgcgcga
     4561 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     4621 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     4681 gggccatgtt aagggcggtt ttttcctgtt tggtcactta ccaatgctta atcagtgagg
     4741 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4801 agataactac gatgcgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4861 acccacgctc accggctcca gatttatcag caataaacca gccagccggg agggccgagc
     4921 gcagaagtga tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4981 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5041 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5101 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5161 tcgttgccag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5221 attctcttac tgtcatgcca tccgtgagat gcttttctgt gactggtgag tactcaacca
     5281 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5341 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5401 ggcgtaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5461 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5521 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcattt
     5581 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     5641 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aagtcaaaag cctccggtcg
     5701 gaggcttttg actttctgct atggaggtca ggtatgattt aaatggtcag ttaga
//
