LOCUS       sequence-297614-        5170 bp    DNA     circular SYN 15-DEC-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 5170)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5170)
  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-m4046301
FEATURES             Location/Qualifiers
     source          1..5170
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             845..862
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             863..880
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(925..943)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1294..2097
                     /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      2180..3162
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3240..3269
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3291..3878
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3890..3917)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3941..4132)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4133..4924,4925..4993))
                     /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     4756..4775
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4994..5096)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5097..5128)
                     /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 atgggtaatg caacagaagt gcctgccaat tcaactgtat tatctttctg
      481 tgcttttgct gtagatgctg ctaaagctta caaagattat ctagctagtg ggggacaacc
      541 aatcactaat tgtgttaaga tgttgtgtac acacactggt actggtcagg caataacagt
      601 tacaccggaa gccaatatgg atcaagaatc ctttggtggt gcatcgtgtt gtctgtactg
      661 ccgttgccac atagatcatc caaatcctaa aggattttgt gacttaaaag gtaagtatgt
      721 acaaatacct acaacttgtg ctaatgaccc tgtgggtttt acacttaaaa acacagtctg
      781 taccgtctgc ggtatgtgga aaggttatgg ctgtagttgt gatcaactcc gcgaacccat
      841 gcttctcgtg ccgagaggtt ctcaccatca tcaccatcat taaacaggcc ccttttcctt
      901 tgtcgatatc atgtaattag ttatgtcacg cttacattca cgccctcctc ccacatccgc
      961 tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttta
     1021 atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac
     1081 aaagcggccg ccgaaccact agtgcactgc agtacacggt ttccttgaaa tttttttgat
     1141 tcggtaatct ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat
     1201 atacgcatat gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac
     1261 agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt
     1321 gctgctactc atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa
     1381 acaaacttgt gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa
     1441 gcattaggtc ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc
     1501 atggagggca cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc
     1561 gaggacagaa aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta
     1621 tacagaatag cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt
     1681 gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg
     1741 ttagcagaat tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt
     1801 gacattgcga agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt
     1861 ggaagagatg aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag
     1921 ggagatgcat tgggtcaaca gtatagaacc gtggatgatg tggtttctac aggatctgac
     1981 attattattg ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa
     2041 cgttacagaa aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa
     2101 actgtattat aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta
     2161 tatcagttat tacccgagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa
     2221 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
     2281 caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa
     2341 tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
     2401 aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac
     2461 aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc
     2521 ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt
     2581 tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc
     2641 ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc
     2701 gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg
     2761 atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta
     2821 cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac
     2881 tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag
     2941 tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca
     3001 gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt
     3061 ttagtagctc gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc
     3121 ttttggtttt caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc
     3181 ccgagcggcc gcgtgttaca agcttgactc cagcgtaact ggactgcaat caactcactg
     3241 gctcaccttc acgggtgggc ctttcttcgg tagaaaatca aaggatcttc ttgagatcct
     3301 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3361 tgtttgccgg atcaagagct accaactctt tttccgaggt aactggcttc agcagagcgc
     3421 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3481 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3541 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3601 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3661 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3721 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3781 taaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcatcgat
     3841 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc
     3901 ccacccgaag gtgagccagg tgattacatt tgggccctca tcagaggttt tcaccgtcat
     3961 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     4021 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     4081 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttaccaat
     4141 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4201 gactccccgt cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg
     4261 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4321 ccgggagggc cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta
     4381 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4441 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4501 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4561 ccttcggtcc tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta
     4621 tggcagcact gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg
     4681 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4741 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4801 gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4861 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4921 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4981 gttgaatact cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc
     5041 ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc
     5101 aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg
     5161 gtcagttaga
//
