LOCUS       sequence-287712-        4923 bp    DNA     circular SYN 15-DEC-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 4923)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4923)
  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-m4046458
FEATURES             Location/Qualifiers
     source          1..4923
                     /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..633
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(678..696)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1047..1850
                     /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      1933..2915
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2993..3022
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3044..3631
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3643..3670)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3694..3885)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3886..4677,4678..4746))
                     /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     4509..4528
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4747..4849)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4850..4881)
                     /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 ggttacataa atgtctttgc attccctttc accatctata gtcttctatt atgcagaatg
      541 aactcccgaa actacattgc ccaggttgac gtagttaact ttaatttgac tctcgtgccg
      601 agaggttctt ggagccatcc gcagtttgaa aaataaacag gccccttttc ctttgtcgat
      661 atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc cgctctaacc
      721 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt ttaatagtta
      781 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacaaagcgg
      841 ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt gattcggtaa
      901 tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca
      961 tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa
     1021 aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta
     1081 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact
     1141 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag
     1201 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg
     1261 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaggaca
     1321 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa
     1381 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg
     1441 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag
     1501 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg
     1561 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag
     1621 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagatg
     1681 cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct gacattatta
     1741 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca
     1801 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat
     1861 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt
     1921 tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga
     1981 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     2041 aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg
     2101 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     2161 acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg
     2221 catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg
     2281 cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga
     2341 aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt
     2401 tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata
     2461 ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc
     2521 ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag
     2581 gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt
     2641 tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat
     2701 gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat
     2761 atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag
     2821 ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt
     2881 tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac ttcccgagcg
     2941 gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca ctggctcacc
     3001 ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat cctttttttc
     3061 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3121 cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag cgcagatacc
     3181 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3241 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3301 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3361 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3421 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3481 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag gggtaaacgc
     3541 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc gatttttgtg
     3601 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa ggcccacccg
     3661 aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt catcaccgaa
     3721 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     3781 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     3841 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc aatgcttaat
     3901 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3961 cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg ctgcaatgat
     4021 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccgggag
     4081 ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4141 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4201 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4261 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4321 tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4381 actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga ctggtgagta
     4441 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4501 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4561 ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4621 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4681 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4741 actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     4801 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc
     4861 tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa atggtcagtt
     4921 aga
//
