LOCUS       sequence-291654-        4834 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 4834)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4834)
  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-m4046652
FEATURES             Location/Qualifiers
     source          1..4834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(589..607)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             958..1761
                     /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      1844..2826
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2904..2933
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2955..3542
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3554..3581)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3605..3796)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3797..4588,4589..4657))
                     /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     4420..4439
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4658..4760)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4761..4792)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atgggttaca taaatgtctt tgcattccct ttcaccatct atagtcttct
      481 attatgcaga atgaactccc gaaactacat tgcccaggtt gacgtagtta actttaattt
      541 gacttaaaca ggcccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca
      601 ttcacgccct cctcccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt
      661 ctaggtccct atttattttt tttaatagtt atgttagtat taagaacgtt atttatattt
      721 caaatttttc ttttttttct gtacaaagcg gccgccgaac cactagtgca ctgcagtaca
      781 cggtttcctt gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag
      841 gagcacagac ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc
      901 ccagtattct taacccaact gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg
      961 tcgaaagcta catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta
     1021 tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc
     1081 aaggaattac tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat
     1141 gtggatatct tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc
     1201 gccaagtaca attttttact cttcgaggac agaaaatttg ctgacattgg taatacagtc
     1261 aaattgcagt actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca
     1321 cacggtgtgg tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca
     1381 aaggaaccta gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact
     1441 ggagaatata ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc
     1501 tttattgctc aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca
     1561 cccggtgtgg gtttagatga caagggagat gcattgggtc aacagtatag aaccgtggat
     1621 gatgtggttt ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga
     1681 agggatgcta aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga
     1741 agatgcggcc agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca
     1801 caaattagag cttcaattta attatatcag ttattacccg agtaacgaag catctgtgct
     1861 tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag
     1921 ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg
     1981 tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     2041 ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga
     2101 atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa
     2161 gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt
     2221 tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc
     2281 ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca
     2341 ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt
     2401 tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt
     2461 cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg
     2521 attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata
     2581 aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg
     2641 taggttatat agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt
     2701 tgtggaagcg gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt
     2761 tttgaaagtg cgtcatcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact
     2821 ttctagctag agaataggaa cttcccgagc ggccgcgtgt tacaagcttg actccagcgt
     2881 aactggactg caatcaactc actggctcac cttcacgggt gggcctttct tcggtagaaa
     2941 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3001 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3061 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     3121 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3181 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3241 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3301 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3361 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3421 agcgcacgag ggagcttcca ggggtaaacg cctggtatct ttatagtcct gtcgggtttc
     3481 gccacctctg acttgagcat cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3541 aaaacgccag caacgcagaa aggcccaccc gaaggtgagc caggtgatta catttgggcc
     3601 ctcatcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     3661 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     3721 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     3781 tttcctgttt ggtcacttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3841 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atgcgggagg
     3901 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3961 atttatcagc aataaaccag ccagccggga gggccgagcg cagaagtgat cctgcaactt
     4021 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4081 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4141 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4201 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgccaga agtaagttgg
     4261 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4321 ccgtgagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4381 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4441 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgtaaactc tcaaggatct
     4501 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4561 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4621 agggaataag ggcgacacgg aaatgttgaa tactcatttt agcttcctta gctcctgaaa
     4681 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     4741 aacctcttac gtgccgatca agtcaaaagc ctccggtcgg aggcttttga ctttctgcta
     4801 tggaggtcag gtatgattta aatggtcagt taga
//
