LOCUS       sequence-287749-        4965 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF10 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4965)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4965)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046236
FEATURES             Location/Qualifiers
     source          1..4965
                     /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             592..609
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             610..675
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(720..738)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1089..1892
                     /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      1975..2957
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3035..3064
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3086..3673
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3685..3712)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3736..3927)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3928..4719,4720..4788))
                     /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     4551..4570
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4789..4891)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4892..4923)
                     /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 ggctatataa acgttttcgc ttttccgttt acgatatata gtctactctt gtgcagaatg
      541 aattctcgta actacatagc acaagtagat gtagttaact ttaatctcac actcgtgccg
      601 agaggttctg attataaaga tcatgatggt gattataaag atcatgatat tgattataaa
      661 gatgatgatg ataaataaac aggccccttt tcctttgtcg atatcatgta attagttatg
      721 tcacgcttac attcacgccc tcctcccaca tccgctctaa ccgaaaagga aggagttaga
      781 caacctgaag tctaggtccc tatttatttt ttttaatagt tatgttagta ttaagaacgt
      841 tatttatatt tcaaattttt cttttttttc tgtacaaagc ggccgccgaa ccactagtgc
      901 actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa cagaaggaag
      961 aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa
     1021 catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc aggaaacgaa
     1081 gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg
     1141 ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg
     1201 ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac
     1261 taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt aagccgctaa
     1321 aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt gctgacattg
     1381 gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa tgggcagaca
     1441 ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcag
     1501 aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct
     1561 ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt
     1621 ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt
     1681 tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt caacagtata
     1741 gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga agaggactat
     1801 ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca ggctgggaag
     1861 catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta aatgcatgta
     1921 tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc gagtaacgaa
     1981 gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca
     2041 aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa
     2101 cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc
     2161 aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt
     2221 accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt
     2281 tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct
     2341 cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta
     2401 ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag
     2461 ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat
     2521 tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt
     2581 atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg
     2641 tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa
     2701 tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa
     2761 aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt
     2821 tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc
     2881 gtttttggtt ttttgaaagt gcgtcatcag agcgcttttg gttttcaaaa gcgctctgaa
     2941 gttcctatac tttctagcta gagaatagga acttcccgag cggccgcgtg ttacaagctt
     3001 gactccagcg taactggact gcaatcaact cactggctca ccttcacggg tgggcctttc
     3061 ttcggtagaa aatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3121 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3181 ctctttttcc gaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3241 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3301 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3361 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3421 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3481 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3541 cggaacagga gagcgcacga gggagcttcc aggggtaaac gcctggtatc tttatagtcc
     3601 tgtcgggttt cgccacctct gacttgagca tcgatttttg tgatgctcgt caggggggcg
     3661 gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc cgaaggtgag ccaggtgatt
     3721 acatttgggc cctcatcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     3781 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     3841 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     3901 aagggcggtt ttttcctgtt tggtcactta ccaatgctta atcagtgagg cacctatctc
     3961 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4021 gatgcgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4081 accggctcca gatttatcag caataaacca gccagccggg agggccgagc gcagaagtga
     4141 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     4201 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4261 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4321 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgccag
     4381 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4441 tgtcatgcca tccgtgagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4501 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4561 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgtaaact
     4621 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4681 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4741 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcattt tagcttcctt
     4801 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     4861 gtgaaagttg gaacctctta cgtgccgatc aagtcaaaag cctccggtcg gaggcttttg
     4921 actttctgct atggaggtca ggtatgattt aaatggtcag ttaga
//
