LOCUS       sequence-307335-        5802 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 5802)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5802)
  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-m4046646
FEATURES             Location/Qualifiers
     source          1..5802
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        50..491
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    50..167
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     462..485
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(1582..1600)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1951..2754
                     /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      2837..3819
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3897..3926
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3948..4535
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4547..4574)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4598..4789)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4790..5581,5582..5650))
                     /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     5413..5432
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5651..5753)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5754..5785)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 ggtatgattt aaatggtcag ttagagcggc cgcttgctat attgaagtac ggattagaag
       61 ccgccgagcg ggcgacagcc ctccgacgga agactctcct ccgtgcgtcc tcgtcttcac
      121 cggtcgcgtt cctgaaacgc agatgtgcct cgcgccgcac tgctccgaac aataaagatt
      181 ctacaatact agcttttatg gttatgaaga ggaaaaattg gcagtaacct ggccccacaa
      241 accttcaaat taacgaatca aattaacaac cataggatga taatgcgatt agttttttag
      301 ccttatttct ggggtaatta atcagcgaag cgatgatttt tgatctatta acagatatat
      361 aaatggaaaa gctgcataac cactttaact aatactttca acattttcag tttgtattac
      421 ttcttattca aatgtcataa aagtatcaac aaaaaattgt taatatacct ctatacttta
      481 acgtcaagga gaaaaaacta taatgttaga aaatgtggct tttaatgttg taaataaggg
      541 acactttgat ggacaacagg gtgaagtacc agtttctatc attaataaca ctgtttacac
      601 aaaagttgat ggtgttgatg tagaattgtt tgaaaataaa acaacattac ctgttaatgt
      661 agcatttgag ctttgggcta agcgcaacat taaaccagta ccagaggtga aaatactcaa
      721 taatttgggt gtggacattg ctgctaatac tgtgatctgg gactacaaaa gagatgctcc
      781 agcacatata tctactattg gtgtttgttc tatgactgac atagccaaga aaccaactga
      841 aacgatttgt gcaccactca ctgtcttttt tgatggtaga gttgatggtc aagtagactt
      901 atttagaaat gcccgtaatg gtgttcttat tacagaaggt agtgttaaag gtttacaacc
      961 atctgtaggt cccaaacaag ctagtcttaa tggagtcaca ttaattggag aagccgtaaa
     1021 aacacagttc aattattata agaaagttga tggtgttgtc caacaattac ctgaaactta
     1081 ctttactcag agtagaaatt tacaagaatt taaacccagg agtcaaatgg aaattgattt
     1141 cttagaatta gctatggatg aattcattga acggtataaa ttagaaggct atgccttcga
     1201 acatatcgtt tatggagatt ttagtcatag tcagttaggt ggtttacatc tactgattgg
     1261 actagctaaa cgttttaagg aatcaccttt tgaattagaa gattttattc ctatggacag
     1321 tacagttaaa aactatttca taacagatgc gcaaacaggt tcatctaagt gtgtgtgttc
     1381 tgttattgat ttattacttg atgattttgt tgaaataata aaatcccaag atttatctgt
     1441 agtttctaag gttgtcaaag tgactattga ctatacagaa atttcattta tgctttggtg
     1501 taaagatggc catgtagaaa cattttaccc aaaattataa acaggcccct tttcctttgt
     1561 cgatatcatg taattagtta tgtcacgctt acattcacgc cctcctccca catccgctct
     1621 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt ttttttaata
     1681 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaaa
     1741 gcggccgccg aaccactagt gcactgcagt acacggtttc cttgaaattt ttttgattcg
     1801 gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata
     1861 cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga
     1921 acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct
     1981 gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca
     2041 aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca
     2101 ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg
     2161 gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgag
     2221 gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac
     2281 agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt
     2341 agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta
     2401 gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac
     2461 attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga
     2521 agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga
     2581 gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg tttctacagg atctgacatt
     2641 attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt
     2701 tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact
     2761 gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat
     2821 cagttattac ccgagtaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc
     2881 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     2941 cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc
     3001 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     3061 tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa
     3121 aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt
     3181 tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag
     3241 aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg
     3301 cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat
     3361 tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg
     3421 attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt
     3481 ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac
     3541 aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt
     3601 agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag
     3661 atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta
     3721 gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcatc agagcgcttt
     3781 tggttttcaa aagcgctctg aagttcctat actttctagc tagagaatag gaacttcccg
     3841 agcggccgcg tgttacaagc ttgactccag cgtaactgga ctgcaatcaa ctcactggct
     3901 caccttcacg ggtgggcctt tcttcggtag aaaatcaaag gatcttcttg agatcctttt
     3961 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4021 ttgccggatc aagagctacc aactcttttt ccgaggtaac tggcttcagc agagcgcaga
     4081 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4141 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4201 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4261 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4321 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4381 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccaggggtaa
     4441 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag catcgatttt
     4501 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgca gaaaggccca
     4561 cccgaaggtg agccaggtga ttacatttgg gccctcatca gaggttttca ccgtcatcac
     4621 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     4681 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     4741 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact taccaatgct
     4801 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4861 tccccgtcgt gtagataact acgatgcggg agggcttacc atctggcccc agtgctgcaa
     4921 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4981 ggagggccga gcgcagaagt gatcctgcaa ctttatccgc ctccatccag tctattaatt
     5041 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5101 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5161 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5221 tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5281 cagcactgca taattctctt actgtcatgc catccgtgag atgcttttct gtgactggtg
     5341 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5401 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5461 aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5521 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5581 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5641 gaatactcat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     5701 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa
     5761 agcctccggt cggaggcttt tgactttctg ctatggaggt ca
//
