LOCUS       sequence-297719-        6871 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 6871)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6871)
  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-m4046314
FEATURES             Location/Qualifiers
     source          1..6871
                     /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"
     primer_bind     complement(2626..2644)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2995..3798
                     /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      3881..4863
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4941..4970
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4992..5579
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5591..5618)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5642..5833)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5834..6625,6626..6694))
                     /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     6457..6476
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6695..6797)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6798..6829)
                     /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 tctaaaatcg aggaaggtaa
     1501 gctagtcatt tggataaatg gagataaggg ttacaacggt ttggcagaag taggcaagaa
     1561 atttgaaaag gacacaggta tcaaggtgac ggttgaacat ccagacaagt tagaagaaaa
     1621 attccctcaa gttgctgcta ctggtgatgg tcccgacatt attttctggg cccacgatcg
     1681 ttttggtgga tatgctcagt caggtttgct ggctgagatc accccagaca aggccttcca
     1741 agataaattg tacccattca cttgggatgc tgtcagatac aacggtaagc ttatagcata
     1801 tccaattgct gttgaagcct tatcgttgat ctacaataag gacttgttac ctaacccacc
     1861 aaagacatgg gaagaaattc cagctttgga taaagagcta aaggccaagg gaaaaagcgc
     1921 tttgatgttt aacttacaag aaccttattt cacctggcca ttaatcgctg ctgacggtgg
     1981 ttacgcattt aaatacgaaa acggtaagta tgacattaag gatgtcggcg ttgataatgc
     2041 tggtgcaaag gctggtttga ccttcttggt tgatttaata aagaataaac acatgaacgc
     2101 tgacactgac tactctatcg ccgaagccgc tttcaacaag ggtgaaactg ctatgaccat
     2161 aaatggtcca tgggcctgga gtaacattga tacttctaaa gtaaactatg gtgtcactgt
     2221 tttgcctaca ttcaaaggcc aaccatccaa gccctttgtc ggtgttctat ccgctggcat
     2281 caatgctgct tctccaaaca aggagctagc taaagaattt ttggaaaatt acttgttaac
     2341 tgacgaaggt ttggaggccg ttaacaagga caagccattg ggtgctgtcg ctttgaaatc
     2401 ttacgaagaa gaattggcta aggatcccag aattgccgct accatggaga acgcccaaaa
     2461 gggtgaaatt atgccaaaca tcccacaaat gtcagccttc tggtacgcag ttaggactgc
     2521 cgtgattaat gctgcttccg gtagacaaac cgtcgacgaa gcattaaaag atgctcaaac
     2581 gtaaacaggc cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc
     2641 acgccctcct cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     2701 ggtccctatt tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa
     2761 atttttcttt tttttctgta caaagcggcc gccgaaccac tagtgcactg cagtacacgg
     2821 tttccttgaa atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag
     2881 cacagactta gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca
     2941 gtattcttaa cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg
     3001 aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt
     3061 aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag
     3121 gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg
     3181 gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc
     3241 aagtacaatt ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa
     3301 ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac
     3361 ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag
     3421 gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga
     3481 gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt
     3541 attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc
     3601 ggtgtgggtt tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat
     3661 gtggtttcta caggatctga cattattatt gttggaagag gactatttgc aaagggaagg
     3721 gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga
     3781 tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa
     3841 attagagctt caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca
     3901 ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg
     3961 catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc
     4021 ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     4081 agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc
     4141 tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca
     4201 tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt
     4261 gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata
     4321 aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt
     4381 tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt
     4441 gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt
     4501 ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt
     4561 cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac
     4621 ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag
     4681 gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt
     4741 ggaagcggta ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt
     4801 gaaagtgcgt catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc
     4861 tagctagaga ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac
     4921 tggactgcaa tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc
     4981 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5041 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg
     5101 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5161 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5221 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5281 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5341 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5401 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5461 gcacgaggga gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5521 acctctgact tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5581 acgccagcaa cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc
     5641 atcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     5701 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     5761 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     5821 cctgtttggt cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5881 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct
     5941 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6001 tatcagcaat aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat
     6061 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6121 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6181 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6241 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg
     6301 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6361 tgagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6421 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6481 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac
     6541 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6601 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6661 gaataagggc gacacggaaa tgttgaatac tcattttagc ttccttagct cctgaaaatc
     6721 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
     6781 ctcttacgtg ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg
     6841 aggtcaggta tgatttaaat ggtcagttag a
//
