LOCUS       sequence-287563-        5803 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 5803)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5803)
  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-m4046288
FEATURES             Location/Qualifiers
     source          1..5803
                     /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             1472..1489
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1490..1513
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1558..1576)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1927..2730
                     /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      2813..3795
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3873..3902
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3924..4511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4523..4550)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4574..4765)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4766..5557,5558..5626))
                     /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     5389..5408
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5627..5729)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5730..5761)
                     /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 atgagcctgg aaaacgttgc cttcaatgta gtgaacaaag gacactttga
      481 tgggcaacag ggtgaggttc ctgtctccat tataaataac accgtttata ctaaggtcga
      541 cggcgttgat gtggaattgt tcgaaaataa gacgacatta cccgtcaacg ttgcatttga
      601 actttgggcg aagcgtaaca tcaaaccagt cccagaagtt aagattttga acaatttggg
      661 tgttgacatc gctgctaata ccgtcatttg ggattacaag agagatgctc cggcccatat
      721 ctcaactata ggtgtctgct cgatgactga cattgctaaa aagccaaccg agactatctg
      781 tgctccatta accgttttct ttgacggtag agtagatggt caagtcgacc tcttcagaaa
      841 cgcaaggaac ggtgttttga ttacagaagg tagtgttaaa ggtctacaac cttctgtagg
      901 tccaaagcaa gcgtccttga acggcgtcac gttgattggc gaagccgtta agactcaatt
      961 caattactat aagaaggtcg atggtgttgt gcaacaatta ccagaaactt acttcactca
     1021 gtctcgcaat ttgcaagaat ttaaaccacg atctcaaatg gaaatcgatt ttcttgaact
     1081 agctatggac gagttcatcg aacgttacaa gttggaaggt tatgcttttg aacacattgt
     1141 ctacggtgac ttctctcatt cccagctggg tggtttacac ttgctaattg gactggctaa
     1201 aagattcaag gaatcaccat tcgagttgga agattttatc cctatggact ctaccgttaa
     1261 aaactacttc atcactgatg ctcaaaccgg ttcgagtaag tgtgtttgtt ccgtcattga
     1321 cttgttacta gatgatttcg tagaaataat taagtctcaa gacttgtcag ttgtttccaa
     1381 ggttgtcaag gtaaccattg actatacgga aatctcattt atgttgtggt gcaaagacgg
     1441 tcatgttgaa accttctacc cgaaattgca actcgtgccg agaggttctt ggagccatcc
     1501 gcagtttgaa aaataaacag gccccttttc ctttgtcgat atcatgtaat tagttatgtc
     1561 acgcttacat tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca
     1621 acctgaagtc taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta
     1681 tttatatttc aaatttttct tttttttctg tacaaagcgg ccgccgaacc actagtgcac
     1741 tgcagtacac ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa
     1801 cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca
     1861 tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga
     1921 taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct
     1981 gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt
     2041 cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta
     2101 aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag
     2161 gcattatccg ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt
     2221 aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt
     2281 acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa
     2341 gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc
     2401 ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt
     2461 gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg
     2521 attatgacac ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga
     2581 accgtggatg atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt
     2641 gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca
     2701 tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata
     2761 ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccga gtaacgaagc
     2821 atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa
     2881 gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg
     2941 aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa
     3001 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac
     3061 caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt
     3121 tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct
     3181 tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt
     3241 ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct
     3301 gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg
     3361 cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat
     3421 gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta
     3481 ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata
     3541 ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag
     3601 gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga gatacttttg
     3661 agcaatgttt gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt
     3721 ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc gctctgaagt
     3781 tcctatactt tctagctaga gaataggaac ttcccgagcg gccgcgtgtt acaagcttga
     3841 ctccagcgta actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt
     3901 cggtagaaaa tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3961 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4021 ctttttccga ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4081 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4141 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4201 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4261 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4321 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4381 gaacaggaga gcgcacgagg gagcttccag gggtaaacgc ctggtatctt tatagtcctg
     4441 tcgggtttcg ccacctctga cttgagcatc gatttttgtg atgctcgtca ggggggcgga
     4501 gcctatggaa aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc aggtgattac
     4561 atttgggccc tcatcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     4621 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     4681 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     4741 gggcggtttt ttcctgtttg gtcacttacc aatgcttaat cagtgaggca cctatctcag
     4801 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4861 tgcgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4921 cggctccaga tttatcagca ataaaccagc cagccgggag ggccgagcgc agaagtgatc
     4981 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5041 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5101 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5161 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgccagaa
     5221 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5281 tcatgccatc cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5341 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5401 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct
     5461 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5521 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5581 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatttta gcttccttag
     5641 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     5701 gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac
     5761 tttctgctat ggaggtcagg tatgatttaa atggtcagtt aga
//
