LOCUS       sequence-287646-        5797 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 5797)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5797)
  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-m4046384
FEATURES             Location/Qualifiers
     source          1..5797
                     /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..1507
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1552..1570)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1921..2724
                     /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      2807..3789
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3867..3896
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3918..4505
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4517..4544)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4568..4759)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4760..5551,5552..5620))
                     /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     5383..5402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5621..5723)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5724..5755)
                     /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 agaggttctc accatcatca
     1501 ccatcattaa acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt
     1561 acattcacgc cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga
     1621 agtctaggtc cctatttatt ttttttaata gttatgttag tattaagaac gttatttata
     1681 tttcaaattt ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt
     1741 acacggtttc cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg
     1801 aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat
     1861 tgcccagtat tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc
     1921 atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag
     1981 ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc
     2041 accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca
     2101 catgtggata tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta
     2161 tccgccaagt acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca
     2221 gtcaaattgc agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat
     2281 gcacacggtg tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta
     2341 acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct
     2401 actggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc
     2461 ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg
     2521 acacccggtg tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg
     2581 gatgatgtgg tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag
     2641 ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg
     2701 agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac
     2761 tcacaaatta gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt
     2821 gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     2881 gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat
     2941 ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     3001 aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa
     3061 agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac
     3121 aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa
     3181 ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct
     3241 tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt
     3301 gcattttttc aagataaagg catccccgat tatattctat accgatgtgg attgcgcata
     3361 ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg
     3421 tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt
     3481 tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag
     3541 ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat
     3601 gggtaggtta tatagggata tagcacagag atatatagca aagagatact tttgagcaat
     3661 gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg
     3721 ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat
     3781 actttctagc tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag
     3841 cgtaactgga ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag
     3901 aaaatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3961 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4021 ccgaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4081 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4141 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4201 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4261 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4321 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4381 gagagcgcac gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt
     4441 ttcgccacct ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat
     4501 ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg
     4561 gccctcatca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     4621 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     4681 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     4741 ttttttcctg tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct
     4801 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg
     4861 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4921 cagatttatc agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa
     4981 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5041 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5101 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5161 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt
     5221 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5281 catccgtgag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5341 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5401 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga
     5461 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5521 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5581 aaaagggaat aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg
     5641 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     5701 tggaacctct tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg
     5761 ctatggaggt caggtatgat ttaaatggtc agttaga
//
