LOCUS       sequence-287588-        5839 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 5839)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5839)
  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-m4046169
FEATURES             Location/Qualifiers
     source          1..5839
                     /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..1549
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1594..1612)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1963..2766
                     /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      2849..3831
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3909..3938
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3960..4547
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4559..4586)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4610..4801)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4802..5593,5594..5662))
                     /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     5425..5444
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5663..5765)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5766..5797)
                     /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 tctgattata aagatcatga
     1501 tggtgattat aaagatcatg atattgatta taaagatgat gatgataaat aaacaggccc
     1561 cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc
     1621 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta
     1681 ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     1741 tttctgtaca aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat
     1801 ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga
     1861 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc
     1921 caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat
     1981 aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac
     2041 gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag
     2101 ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact
     2161 gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt
     2221 ttactcttcg aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct
     2281 gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc
     2341 ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc
     2401 cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag
     2461 ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga
     2521 gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta
     2581 gatgacaagg gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca
     2641 ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta
     2701 gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa
     2761 aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca
     2821 atttaattat atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca
     2881 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     2941 acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta
     3001 aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     3061 acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt
     3121 ttgttctaca aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact
     3181 ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc
     3241 cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga
     3301 ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa
     3361 ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg
     3421 atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc
     3481 tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat
     3541 agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta
     3601 gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga
     3661 tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt
     3721 cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca
     3781 tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat
     3841 aggaacttcc cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc
     3901 aactcactgg ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct
     3961 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4021 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca
     4081 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4141 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4201 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4261 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4321 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4381 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4441 ttccaggggt aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4501 agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4561 cagaaaggcc cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt
     4621 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     4681 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     4741 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     4801 cttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4861 tagttgcctg actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc
     4921 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4981 accagccagc cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc
     5041 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5101 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5161 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5221 cggttagctc cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac
     5281 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt
     5341 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5401 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5461 tcatcattgg aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat
     5521 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5581 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5641 cacggaaatg ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa
     5701 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     5761 gatcaagtca aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg
     5821 atttaaatgg tcagttaga
//
