LOCUS       Exported                5850 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 endoRNAse.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046424
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5850)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5850)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5850
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             1519..1536
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1537..1560
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1605..1623)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1974..2777
                     /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      2860..3842
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3920..3949
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3971..4558
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4570..4597
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4621..4812)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4813..5673)
                     /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     5436..5455
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5674..5776)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5777..5808
                     /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 agcctggaaa acgttgcctt caatgtagtg aacaaaggac actttgatgg gcaacagggt
      541 gaggttcctg tctccattat aaataacacc gtttatacta aggtcgacgg cgttgatgtg
      601 gaattgttcg aaaataagac gacattaccc gtcaacgttg catttgaact ttgggcgaag
      661 cgtaacatca aaccagtccc agaagttaag attttgaaca atttgggtgt tgacatcgct
      721 gctaataccg tcatttggga ttacaagaga gatgctccgg cccatatctc aactataggt
      781 gtctgctcga tgactgacat tgctaaaaag ccaaccgaga ctatctgtgc tccattaacc
      841 gttttctttg acggtagagt agatggtcaa gtcgacctct tcagaaacgc aaggaacggt
      901 gttttgatta cagaaggtag tgttaaaggt ctacaacctt ctgtaggtcc aaagcaagcg
      961 tccttgaacg gcgtcacgtt gattggcgaa gccgttaaga ctcaattcaa ttactataag
     1021 aaggtcgatg gtgttgtgca acaattacca gaaacttact tcactcagtc tcgcaatttg
     1081 caagaattta aaccacgatc tcaaatggaa atcgattttc ttgaactagc tatggacgag
     1141 ttcatcgaac gttacaagtt ggaaggttat gcttttgaac acattgtcta cggtgacttc
     1201 tctcattccc agctgggtgg tttacacttg ctaattggac tggctaaaag attcaaggaa
     1261 tcaccattcg agttggaaga ttttatccct atggactcta ccgttaaaaa ctacttcatc
     1321 actgatgctc aaaccggttc gagtaagtgt gtttgttccg tcattgactt gttactagat
     1381 gatttcgtag aaataattaa gtctcaagac ttgtcagttg tttccaaggt tgtcaaggta
     1441 accattgact atacggaaat ctcatttatg ttgtggtgca aagacggtca tgttgaaacc
     1501 ttctacccga aattgcaact cgtgccgaga ggttcttgga gccatccgca gtttgaaaaa
     1561 taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg cttacattca
     1621 cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
     1681 gtccctattt atttttttta atagttatgt tagtattaag aacgttattt atatttcaaa
     1741 tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc agtacacggt
     1801 ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga aggaaggagc
     1861 acagacttag attggtatat atacgcatat gtagtgttga agaaacatga aattgcccag
     1921 tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga
     1981 aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta
     2041 atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg
     2101 aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg
     2161 atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca
     2221 agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat acagtcaaat
     2281 tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg
     2341 gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg
     2401 aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag
     2461 aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta
     2521 ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg
     2581 gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc gtggatgatg
     2641 tggtttctac aggatctgac attattattg ttggaagagg actatttgca aagggaaggg
     2701 atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat
     2761 gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa
     2821 ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc tgtgcttcat
     2881 tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     2941 atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct
     3001 tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     3061 gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct
     3121 atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat
     3181 cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg
     3241 cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa
     3301 aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt
     3361 ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg
     3421 aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc
     3481 tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc
     3541 actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca
     3601 taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg
     3661 ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg
     3721 gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt tggttttttg
     3781 aaagtgcgtc atcagagcgc ttttggtttt caaaagcgct ctgaagttcc tatactttct
     3841 agctagagaa taggaacttc ccgagcggcc gcgtgttaca agcttgactc cagcgtaact
     3901 ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg tagaaaatca
     3961 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4021 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaggt
     4081 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4141 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4201 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4261 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4321 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4381 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4441 cacgagggag cttccagggg taaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4501 cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4561 cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt tgggccctca
     4621 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     4681 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     4741 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     4801 ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4861 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc gggagggctt
     4921 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4981 atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg caactttatc
     5041 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5101 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5161 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5221 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta agttggccgc
     5281 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5341 gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5401 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5461 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc
     5521 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5581 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5641 aataagggcg acacggaaat gttgaatact cattttagct tccttagctc ctgaaaatct
     5701 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     5761 tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga
     5821 ggtcaggtat gatttaaatg gtcagttaga
//
