LOCUS       sequence-287677-        4917 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 4917)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4917)
  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-m4046254
FEATURES             Location/Qualifiers
     source          1..4917
                     /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..627
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(672..690)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1041..1844
                     /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      1927..2909
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2987..3016
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3038..3625
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3637..3664)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3688..3879)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3880..4671,4672..4740))
                     /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     4503..4522
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4741..4843)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4844..4875)
                     /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 agaggttctc accatcatca ccatcattaa acaggcccct tttcctttgt cgatatcatg
      661 taattagtta tgtcacgctt acattcacgc cctcctccca catccgctct aaccgaaaag
      721 gaaggagtta gacaacctga agtctaggtc cctatttatt ttttttaata gttatgttag
      781 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaaa gcggccgccg
      841 aaccactagt gcactgcagt acacggtttc cttgaaattt ttttgattcg gtaatctccg
      901 aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata cgcatatgta
      961 gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga acaaaaacgt
     1021 gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct gctactcatc
     1081 ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca aacttgtgtg
     1141 cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca ttaggtccca
     1201 aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg gagggcacag
     1261 ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgag gacagaaaat
     1321 ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac agaatagcag
     1381 aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt agcggtttga
     1441 agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta gcagaattgt
     1501 catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac attgcgaaga
     1561 gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga agagatgaag
     1621 gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga gatgcattgg
     1681 gtcaacagta tagaaccgtg gatgatgtgg tttctacagg atctgacatt attattgttg
     1741 gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag
     1801 caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact gtattataag
     1861 taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat cagttattac
     1921 ccgagtaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta
     1981 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg
     2041 ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag
     2101 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     2161 gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc
     2221 gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct
     2281 ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta
     2341 ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga
     2401 ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat tatattctat
     2461 accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt
     2521 ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg
     2581 tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg
     2641 tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt
     2701 tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca
     2761 aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt
     2821 acagtccggt gcgtttttgg ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa
     2881 aagcgctctg aagttcctat actttctagc tagagaatag gaacttcccg agcggccgcg
     2941 tgttacaagc ttgactccag cgtaactgga ctgcaatcaa ctcactggct caccttcacg
     3001 ggtgggcctt tcttcggtag aaaatcaaag gatcttcttg agatcctttt tttctgcgcg
     3061 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3121 aagagctacc aactcttttt ccgaggtaac tggcttcagc agagcgcaga taccaaatac
     3181 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3241 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3301 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3361 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3421 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3481 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccaggggtaa acgcctggta
     3541 tctttatagt cctgtcgggt ttcgccacct ctgacttgag catcgatttt tgtgatgctc
     3601 gtcagggggg cggagcctat ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg
     3661 agccaggtga ttacatttgg gccctcatca gaggttttca ccgtcatcac cgaaacgcgc
     3721 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     3781 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     3841 gcgggccatg ttaagggcgg ttttttcctg tttggtcact taccaatgct taatcagtga
     3901 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3961 gtagataact acgatgcggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4021 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg ggagggccga
     4081 gcgcagaagt gatcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4141 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4201 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4261 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4321 gatcgttgcc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4381 taattctctt actgtcatgc catccgtgag atgcttttct gtgactggtg agtactcaac
     4441 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4501 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4561 ggggcgtaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4621 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4681 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4741 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     4801 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa agcctccggt
     4861 cggaggcttt tgactttctg ctatggaggt caggtatgat ttaaatggtc agttaga
//
