LOCUS       sequence-292522-        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-m4046324
FEATURES             Location/Qualifiers
     source          1..4923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        32..473
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    32..149
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     444..467
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             599..616
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             617..640
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(685..703)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1054..1857
                     /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      1940..2922
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3000..3029
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3051..3638
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3650..3677)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3701..3892)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3893..4684,4685..4753))
                     /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     4516..4535
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4754..4856)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4857..4888)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 agttagagcg gccgcttgct atattgaagt acggattaga agccgccgag cgggcgacag
       61 ccctccgacg gaagactctc ctccgtgcgt cctcgtcttc accggtcgcg ttcctgaaac
      121 gcagatgtgc ctcgcgccgc actgctccga acaataaaga ttctacaata ctagctttta
      181 tggttatgaa gaggaaaaat tggcagtaac ctggccccac aaaccttcaa attaacgaat
      241 caaattaaca accataggat gataatgcga ttagtttttt agccttattt ctggggtaat
      301 taatcagcga agcgatgatt tttgatctat taacagatat ataaatggaa aagctgcata
      361 accactttaa ctaatacttt caacattttc agtttgtatt acttcttatt caaatgtcat
      421 aaaagtatca acaaaaaatt gttaatatac ctctatactt taacgtcaag gagaaaaaac
      481 tataatgggc tatataaacg ttttcgcttt tccgtttacg atatatagtc tactcttgtg
      541 cagaatgaat tctcgtaact acatagcaca agtagatgta gttaacttta atctcacact
      601 cgtgccgaga ggttcttgga gccatccgca gtttgaaaaa taaacaggcc ccttttcctt
      661 tgtcgatatc atgtaattag ttatgtcacg cttacattca cgccctcctc ccacatccgc
      721 tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttta
      781 atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac
      841 aaagcggccg ccgaaccact agtgcactgc agtacacggt ttccttgaaa tttttttgat
      901 tcggtaatct ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat
      961 atacgcatat gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac
     1021 agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt
     1081 gctgctactc atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa
     1141 acaaacttgt gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa
     1201 gcattaggtc ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc
     1261 atggagggca cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc
     1321 gaggacagaa aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta
     1381 tacagaatag cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt
     1441 gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg
     1501 ttagcagaat tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt
     1561 gacattgcga agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt
     1621 ggaagagatg aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag
     1681 ggagatgcat tgggtcaaca gtatagaacc gtggatgatg tggtttctac aggatctgac
     1741 attattattg ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa
     1801 cgttacagaa aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa
     1861 actgtattat aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta
     1921 tatcagttat tacccgagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa
     1981 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
     2041 caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa
     2101 tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
     2161 aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac
     2221 aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc
     2281 ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt
     2341 tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc
     2401 ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc
     2461 gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg
     2521 atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta
     2581 cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac
     2641 tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag
     2701 tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca
     2761 gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt
     2821 ttagtagctc gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc
     2881 ttttggtttt caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc
     2941 ccgagcggcc gcgtgttaca agcttgactc cagcgtaact ggactgcaat caactcactg
     3001 gctcaccttc acgggtgggc ctttcttcgg tagaaaatca aaggatcttc ttgagatcct
     3061 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3121 tgtttgccgg atcaagagct accaactctt tttccgaggt aactggcttc agcagagcgc
     3181 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3241 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3301 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3361 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3421 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3481 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3541 taaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcatcgat
     3601 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc
     3661 ccacccgaag gtgagccagg tgattacatt tgggccctca tcagaggttt tcaccgtcat
     3721 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     3781 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     3841 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttaccaat
     3901 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3961 gactccccgt cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg
     4021 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4081 ccgggagggc cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta
     4141 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4201 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4261 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4321 ccttcggtcc tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta
     4381 tggcagcact gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg
     4441 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4501 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4561 gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4621 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4681 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4741 gttgaatact cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc
     4801 ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc
     4861 aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg
     4921 gtc
//
