LOCUS       sequence-291573-        6387 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 3'-to-5' exonuclease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6387)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6387)
  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-m4046374
FEATURES             Location/Qualifiers
     source          1..6387
                     /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"
     CDS             2062..2079
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2080..2097
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2142..2160)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2511..3314
                     /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      3397..4379
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4457..4486
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4508..5095
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5107..5134)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5158..5349)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5350..6141,6142..6210))
                     /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     5973..5992
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6211..6313)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6314..6345)
                     /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 gccgagaatg tcacaggcct attcaaggat tgctcgaaag tgatcaccgg tttacaccct
      541 acgcaggctc caactcattt gtccgttgac accaagttta agactgaagg attgtgtgtt
      601 gacataccag gtattcccaa agatatgact taccgtaggc tgatttctat gatgggtttc
      661 aagatgaact atcaagttaa cggttaccca aatatgttta tcaccagaga agaagctatt
      721 agacacgtac gtgcatggat cggtttcgat gtcgaaggtt gtcatgctac tagagaagcg
      781 gtcggtacaa acttgccatt acaacttggc ttttcaaccg gtgttaattt ggttgctgtt
      841 cctacagggt atgtcgacac tccaaacaac acggatttct ctcgagtgtc cgccaagcca
      901 ccacctggtg atcaattcaa acacttgatc ccgctcatgt acaagggttt gccttggaac
      961 gttgtcagaa ttaaaatagt acaaatgtta tctgatactt tgaagaatct gagtgacaga
     1021 gttgtcttcg tcctttgggc tcacggtttt gagctaacca gcatgaagta ctttgtgaag
     1081 attggaccag aacggacttg ttgcttgtgt gacagaagag ccacttgttt ctccaccgct
     1141 agcgatacat atgcatgttg gcatcattct attggtttcg actacgttta caaccccttt
     1201 atgatcgacg ttcaacagtg gggtttcacc ggtaatttgc aatcgaacca cgacttgtat
     1261 tgtcaagttc acggcaatgc tcacgtggct tcatgcgatg ctattatgac aaggtgttta
     1321 gccgtccatg aatgtttcgt taaaagagta gactggacta tcgaataccc aattatcggt
     1381 gatgaactca agattaacgc tgcctgtaga aaggtccaac atatggttgt aaaggcagct
     1441 ctattagctg acaaatttcc agttttgcac gatataggga atccaaaggc cataaagtgt
     1501 gttccacaag ctgacgtcga atggaaattc tacgatgcgc aaccctgctc tgataaagct
     1561 tataagatcg aggaattgtt ttactcttac gctacccact ccgataagtt cactgacggt
     1621 gtgtgtcttt tctggaactg caatgttgat agatatccag caaactccat cgtttgtaga
     1681 ttcgacacgc gagtcttgag caacttgaac ttaccaggtt gtgatggtgg ttccttgtac
     1741 gtcaataaac atgcatttca cactcccgct ttcgacaagt ctgcttttgt taatttaaag
     1801 caattgcctt ttttctatta ctctgatagt ccatgcgaat ctcatggtaa gcaagtcgtt
     1861 tccgacattg attacgtacc actgaagtca gccacctgta taacaagatg taaccttgga
     1921 ggtgctgtct gccgccacca tgctaacgag tacagactat acttggatgc ctataacatg
     1981 atgatttcag ccggtttctc tttatgggtt tacaagcagt ttgacactta caacttgtgg
     2041 aacaccttca cgcgtttgca actcgtgccg agaggttctc accatcatca ccatcattaa
     2101 acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc
     2161 cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
     2221 cctatttatt ttttttaata gttatgttag tattaagaac gttatttata tttcaaattt
     2281 ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt acacggtttc
     2341 cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
     2401 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
     2461 tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag
     2521 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
     2581 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
     2641 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
     2701 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
     2761 acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca gtcaaattgc
     2821 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
     2881 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
     2941 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
     3001 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
     3061 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     3121 tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     3181 tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     3241 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     3301 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     3361 gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt gcttcatttt
     3421 gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     3481 tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca
     3541 tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     3601 gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata
     3661 cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt
     3721 agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac
     3781 tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa
     3841 aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc
     3901 aagataaagg catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac
     3961 agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat
     4021 tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact
     4081 ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa
     4141 aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta
     4201 tatagggata tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa
     4261 gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa
     4321 gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctagc
     4381 tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag cgtaactgga
     4441 ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag aaaatcaaag
     4501 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4561 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaggtaac
     4621 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4681 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4741 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4801 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4861 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4921 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4981 gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5041 ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5101 cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg gccctcatca
     5161 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     5221 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     5281 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     5341 tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5401 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg agggcttacc
     5461 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5521 agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa ctttatccgc
     5581 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5641 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5701 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5761 caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt
     5821 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtgag
     5881 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5941 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6001 aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct
     6061 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6121 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6181 aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg aaaatctcga
     6241 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     6301 tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg ctatggaggt
     6361 caggtatgat ttaaatggtc agttaga
//
