LOCUS       sequence-482818-        6918 bp    DNA     circular SYN 19-AUG-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 6918)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6918)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 19, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046159
FEATURES             Location/Qualifiers
     source          1..6918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1513..1530
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2673..2691)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3042..3845
                     /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      3928..4910
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4988..5017
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5039..5626
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5638..5665)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5689..5880)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5881..6672,6673..6741))
                     /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     6504..6523
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6742..6844)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6845..6876)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 ttagaaaatg tggcttttaa tgttgtaaat aagggacact ttgatggaca acagggtgaa
      541 gtaccagttt ctatcattaa taacactgtt tacacaaaag ttgatggtgt tgatgtagaa
      601 ttgtttgaaa ataaaacaac attacctgtt aatgtagcat ttgagctttg ggctaagcgc
      661 aacattaaac cagtaccaga ggtgaaaata ctcaataatt tgggtgtgga cattgctgct
      721 aatactgtga tctgggacta caaaagagat gctccagcac atatatctac tattggtgtt
      781 tgttctatga ctgacatagc caagaaacca actgaaacga tttgtgcacc actcactgtc
      841 ttttttgatg gtagagttga tggtcaagta gacttattta gaaatgcccg taatggtgtt
      901 cttattacag aaggtagtgt taaaggttta caaccatctg taggtcccaa acaagctagt
      961 cttaatggag tcacattaat tggagaagcc gtaaaaacac agttcaatta ttataagaaa
     1021 gttgatggtg ttgtccaaca attacctgaa acttacttta ctcagagtag aaatttacaa
     1081 gaatttaaac ccaggagtca aatggaaatt gatttcttag aattagctat ggatgaattc
     1141 attgaacggt ataaattaga aggctatgcc ttcgaacata tcgtttatgg agattttagt
     1201 catagtcagt taggtggttt acatctactg attggactag ctaaacgttt taaggaatca
     1261 ccttttgaat tagaagattt tattcctatg gacagtacag ttaaaaacta tttcataaca
     1321 gatgcgcaaa caggttcatc taagtgtgtg tgttctgtta ttgatttatt acttgatgat
     1381 tttgttgaaa taataaaatc ccaagattta tctgtagttt ctaaggttgt caaagtgact
     1441 attgactata cagaaatttc atttatgctt tggtgtaaag atggccatgt agaaacattt
     1501 tacccaaaat tactcgtgcc gagaggttct aaaatcgagg aaggtaagct agtcatttgg
     1561 ataaatggag ataagggtta caacggtttg gcagaagtag gcaagaaatt tgaaaaggac
     1621 acaggtatca aggtgacggt tgaacatcca gacaagttag aagaaaaatt ccctcaagtt
     1681 gctgctactg gtgatggtcc cgacattatt ttctgggccc acgatcgttt tggtggatat
     1741 gctcagtcag gtttgctggc tgagatcacc ccagacaagg ccttccaaga taaattgtac
     1801 ccattcactt gggatgctgt cagatacaac ggtaagctta tagcatatcc aattgctgtt
     1861 gaagccttat cgttgatcta caataaggac ttgttaccta acccaccaaa gacatgggaa
     1921 gaaattccag ctttggataa agagctaaag gccaagggaa aaagcgcttt gatgtttaac
     1981 ttacaagaac cttatttcac ctggccatta atcgctgctg acggtggtta cgcatttaaa
     2041 tacgaaaacg gtaagtatga cattaaggat gtcggcgttg ataatgctgg tgcaaaggct
     2101 ggtttgacct tcttggttga tttaataaag aataaacaca tgaacgctga cactgactac
     2161 tctatcgccg aagccgcttt caacaagggt gaaactgcta tgaccataaa tggtccatgg
     2221 gcctggagta acattgatac ttctaaagta aactatggtg tcactgtttt gcctacattc
     2281 aaaggccaac catccaagcc ctttgtcggt gttctatccg ctggcatcaa tgctgcttct
     2341 ccaaacaagg agctagctaa agaatttttg gaaaattact tgttaactga cgaaggtttg
     2401 gaggccgtta acaaggacaa gccattgggt gctgtcgctt tgaaatctta cgaagaagaa
     2461 ttggctaagg atcccagaat tgccgctacc atggagaacg cccaaaaggg tgaaattatg
     2521 ccaaacatcc cacaaatgtc agccttctgg tacgcagtta ggactgccgt gattaatgct
     2581 gcttccggta gacaaaccgt cgacgaagca ttaaaagatg ctcaaacgta aacaggcccc
     2641 ttttcctttg tcgatatcat gtaattagtt atgtcacgct tacattcacg ccctcctccc
     2701 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
     2761 tttttttaat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     2821 ttctgtacaa agcggccgcc gaaccactag tgcactgcag tacacggttt ccttgaaatt
     2881 tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat
     2941 tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc
     3001 aactgcacag aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa gctacatata
     3061 aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg
     3121 aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt
     3181 tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg
     3241 atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt
     3301 tactcttcga ggacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg
     3361 cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc
     3421 caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc
     3481 ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg
     3541 gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag
     3601 acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag
     3661 atgacaaggg agatgcattg ggtcaacagt atagaaccgt ggatgatgtg gtttctacag
     3721 gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag
     3781 agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa
     3841 actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa
     3901 tttaattata tcagttatta cccgagtaac gaagcatctg tgcttcattt tgtagaacaa
     3961 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     4021 cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa
     4081 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     4141 cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt
     4201 tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt
     4261 tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc
     4321 gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac
     4381 tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag
     4441 gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga
     4501 tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct
     4561 atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata
     4621 gttcttacta caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag
     4681 aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat
     4741 atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc
     4801 gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtcat
     4861 cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag ctagagaata
     4921 ggaacttccc gagcggccgc gtgttacaag cttgactcca gcgtaactgg actgcaatca
     4981 actcactggc tcaccttcac gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt
     5041 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5101 cggtggtttg tttgccggat caagagctac caactctttt tccgaggtaa ctggcttcag
     5161 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     5221 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5281 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5341 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5401 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5461 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5521 tccaggggta aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5581 gcatcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5641 agaaaggccc acccgaaggt gagccaggtg attacatttg ggccctcatc agaggttttc
     5701 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     5761 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     5821 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     5881 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5941 agttgcctga ctccccgtcg tgtagataac tacgatgcgg gagggcttac catctggccc
     6001 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6061 ccagccagcc gggagggccg agcgcagaag tgatcctgca actttatccg cctccatcca
     6121 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6181 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6241 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6301 ggttagctcc ttcggtcctc cgatcgttgc cagaagtaag ttggccgcag tgttatcact
     6361 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtga gatgcttttc
     6421 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6481 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6541 catcattgga aaacgttctt cggggcgtaa actctcaagg atcttaccgc tgttgagatc
     6601 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6661 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6721 acggaaatgt tgaatactca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     6781 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
     6841 atcaagtcaa aagcctccgg tcggaggctt ttgactttct gctatggagg tcaggtatga
     6901 tttaaatggt cagttaga
//
