LOCUS       sequence-298363-        5181 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5181)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5181)
  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-m4046636
FEATURES             Location/Qualifiers
     source          1..5181
                     /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"
     primer_bind     complement(936..954)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1305..2108
                     /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      2191..3173
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3251..3280
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3302..3889
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3901..3928)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3952..4143)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4144..4935,4936..5004))
                     /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     4767..4786
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5005..5107)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5108..5139)
                     /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 ggtaatgcaa cagaagtgcc tgccaattca actgtattat ctttctgtgc ttttgctgta
      541 gatgctgcta aagcttacaa agattatcta gctagtgggg gacaaccaat cactaattgt
      601 gttaagatgt tgtgtacaca cactggtact ggtcaggcaa taacagttac accggaagcc
      661 aatatggatc aagaatcctt tggtggtgca tcgtgttgtc tgtactgccg ttgccacata
      721 gatcatccaa atcctaaagg attttgtgac ttaaaaggta agtatgtaca aatacctaca
      781 acttgtgcta atgaccctgt gggttttaca cttaaaaaca cagtctgtac cgtctgcggt
      841 atgtggaaag gttatggctg tagttgtgat caactccgcg aacccatgct ttaaacaggc
      901 cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc acgccctcct
      961 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
     1021 tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt
     1081 tttttctgta caaagcggcc gccgaaccac tagtgcactg cagtacacgg tttccttgaa
     1141 atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta
     1201 gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa
     1261 cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg aaagctacat
     1321 ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt aatatcatgc
     1381 acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag gaattactgg
     1441 agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg gatatcttga
     1501 ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc aagtacaatt
     1561 ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa ttgcagtact
     1621 ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac ggtgtggtgg
     1681 gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag gaacctagag
     1741 gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga gaatatacta
     1801 agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt attgctcaaa
     1861 gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc ggtgtgggtt
     1921 tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat gtggtttcta
     1981 caggatctga cattattatt gttggaagag gactatttgc aaagggaagg gatgctaagg
     2041 tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga tgcggccagc
     2101 aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa attagagctt
     2161 caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa
     2221 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     2281 gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg
     2341 taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt
     2401 ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt
     2461 ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta
     2521 ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg
     2581 tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct
     2641 gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata
     2701 aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag
     2761 tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc
     2821 tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga
     2881 atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg
     2941 tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg
     3001 gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta
     3061 ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt
     3121 catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc tagctagaga
     3181 ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac tggactgcaa
     3241 tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc aaaggatctt
     3301 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3361 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg taactggctt
     3421 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3481 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3541 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3601 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3661 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3721 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3781 gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3841 tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3901 cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc atcagaggtt
     3961 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     4021 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     4081 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     4141 cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4201 catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct taccatctgg
     4261 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4321 aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat ccgcctccat
     4381 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4441 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4501 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4561 agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg cagtgttatc
     4621 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg tgagatgctt
     4681 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4741 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4801 gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac cgctgttgag
     4861 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4921 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4981 gacacggaaa tgttgaatac tcattttagc ttccttagct cctgaaaatc tcgataactc
     5041 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     5101 ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg aggtcaggta
     5161 tgatttaaat ggtcagttag a
//
