LOCUS       sequence-307327-        5056 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5056)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5056)
  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-m4046630
FEATURES             Location/Qualifiers
     source          1..5056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(795..813)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1164..1967
                     /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      2050..3032
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3110..3139
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3161..3748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3760..3787)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3811..4002)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4003..4794,4795..4863))
                     /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     4626..4645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4864..4966)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4967..4998)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        join(5050..5056,1..401)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 ccatagcttc aaaatgtttc tactcctttt ttactcttcc agattttctc ggactccgcg
       61 catcgccgta ccacttcaaa acacccaagc acagcatact aaatttcccc tctttcttcc
      121 tctagggtgt cgttaattac ccgtactaaa ggtttggaaa agaaaaaaga gaccgcctcg
      181 tttctttttc ttcgtcgaaa aaggcaataa aaatttttat cacgtttctt tttcttgaaa
      241 attttttttt ttgatttttt tctctttcga tgacctccca ttgatattta agttaataaa
      301 cggtcttcaa tttctcaagt ttcagtttca tttttcttgt tctattacaa ctttttttac
      361 ttcttgctca ttagaaagaa agcatagcaa tctaatctaa gttttaatta caaaatgaat
      421 gagcttagtc ctgttgcact acgacagatg tcttgtgctg ccggtactac acaaactgct
      481 tgcactgatg acaatgcgtt agcttactac aacacaacaa agggaggtag gtttgtactt
      541 gcactgttat ccgatttaca ggatttgaaa tgggctagat tccctaagag tgatggaact
      601 ggtactatct atacagaact ggaaccacct tgtaggtttg ttacagacac acctaaaggt
      661 cctaaagtga agtatttata ctttattaaa ggattaaaca acctaaatag aggtatggta
      721 cttggtagtt tagctgccac agtacgtcta taaacaggcc ccttttcctt tgtcgatatc
      781 atgtaattag ttatgtcacg cttacattca cgccctcctc ccacatccgc tctaaccgaa
      841 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttta atagttatgt
      901 tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac aaagcggccg
      961 ccgaaccact agtgcactgc agtacacggt ttccttgaaa tttttttgat tcggtaatct
     1021 ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat
     1081 gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa
     1141 cgtgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc
     1201 atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt
     1261 gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc
     1321 ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca
     1381 cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc gaggacagaa
     1441 aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag
     1501 cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt
     1561 tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat
     1621 tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga
     1681 agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg
     1741 aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagatgcat
     1801 tgggtcaaca gtatagaacc gtggatgatg tggtttctac aggatctgac attattattg
     1861 ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa
     1921 aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat
     1981 aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat
     2041 tacccgagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg
     2101 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaaa
     2161 gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga
     2221 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     2281 cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac aaaaatgcat
     2341 cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc ctttgtgcgc
     2401 tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt tagaagaagg
     2461 ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc ccgcgtttac
     2521 tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc gattatattc
     2581 tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg atgattcttc
     2641 attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta cgtataggaa
     2701 atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac tacaattttt
     2761 ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag tttagatgca
     2821 agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca gagatatata
     2881 gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc
     2941 gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc ttttggtttt
     3001 caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc ccgagcggcc
     3061 gcgtgttaca agcttgactc cagcgtaact ggactgcaat caactcactg gctcaccttc
     3121 acgggtgggc ctttcttcgg tagaaaatca aaggatcttc ttgagatcct ttttttctgc
     3181 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3241 atcaagagct accaactctt tttccgaggt aactggcttc agcagagcgc agataccaaa
     3301 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3361 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3421 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3481 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3541 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3601 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg taaacgcctg
     3661 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcatcgat ttttgtgatg
     3721 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag
     3781 gtgagccagg tgattacatt tgggccctca tcagaggttt tcaccgtcat caccgaaacg
     3841 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     3901 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     3961 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttaccaat gcttaatcag
     4021 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4081 cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg caatgatacc
     4141 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccgggagggc
     4201 cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4261 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4321 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4381 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4441 tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4501 gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg gtgagtactc
     4561 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4621 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4681 ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4741 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4801 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4861 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
     4921 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc
     4981 ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg gtcagttaga
     5041 gcggccgccc cacaca
//
