LOCUS       sequence-287647-        5791 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 5791)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5791)
  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-m4046205
FEATURES             Location/Qualifiers
     source          1..5791
                     /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             1466..1483
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1484..1501
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1546..1564)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1915..2718
                     /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      2801..3783
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3861..3890
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3912..4499
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4511..4538)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4562..4753)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4754..5545,5546..5614))
                     /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     5377..5396
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5615..5717)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5718..5749)
                     /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 aattactcgt gccgagaggt tctcaccatc atcaccatca
     1501 ttaaacaggc cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc
     1561 acgccctcct cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     1621 ggtccctatt tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa
     1681 atttttcttt tttttctgta caaagcggcc gccgaaccac tagtgcactg cagtacacgg
     1741 tttccttgaa atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag
     1801 cacagactta gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca
     1861 gtattcttaa cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg
     1921 aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt
     1981 aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag
     2041 gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg
     2101 gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc
     2161 aagtacaatt ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa
     2221 ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac
     2281 ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag
     2341 gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga
     2401 gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt
     2461 attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc
     2521 ggtgtgggtt tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat
     2581 gtggtttcta caggatctga cattattatt gttggaagag gactatttgc aaagggaagg
     2641 gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga
     2701 tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa
     2761 attagagctt caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca
     2821 ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg
     2881 catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc
     2941 ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     3001 agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc
     3061 tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca
     3121 tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt
     3181 gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata
     3241 aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt
     3301 tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt
     3361 gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt
     3421 ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt
     3481 cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac
     3541 ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag
     3601 gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt
     3661 ggaagcggta ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt
     3721 gaaagtgcgt catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc
     3781 tagctagaga ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac
     3841 tggactgcaa tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc
     3901 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3961 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg
     4021 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4081 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4141 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4201 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4261 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4321 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4381 gcacgaggga gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4441 acctctgact tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4501 acgccagcaa cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc
     4561 atcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     4621 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     4681 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     4741 cctgtttggt cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4801 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct
     4861 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4921 tatcagcaat aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat
     4981 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5041 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5101 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5161 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg
     5221 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5281 tgagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5341 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5401 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac
     5461 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5521 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5581 gaataagggc gacacggaaa tgttgaatac tcattttagc ttccttagct cctgaaaatc
     5641 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
     5701 ctcttacgtg ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg
     5761 aggtcaggta tgatttaaat ggtcagttag a
//
