LOCUS       Exported                5886 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 endoRNAse.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046190
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5886)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5886)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5886
                     /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             1513..1530
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1531..1596
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1641..1659)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2010..2813
                     /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      2896..3878
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3956..3985
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4007..4594
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4606..4633
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4657..4848)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4849..5709)
                     /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     5472..5491
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5710..5812)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5813..5844
                     /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 ttagaaaatg tggcttttaa tgttgtaaat aagggacact ttgatggaca acagggtgaa
      541 gtaccagttt ctatcattaa taacactgtt tacacaaaag ttgatggtgt tgatgtagaa
      601 ttgtttgaaa ataaaacaac attacctgtt aatgtagcat ttgagctttg ggctaagcgc
      661 aacattaaac cagtaccaga ggtgaaaata ctcaataatt tgggtgtgga cattgctgct
      721 aatactgtga tctgggacta caaaagagat gctccagcac atatatctac tattggtgtt
      781 tgttctatga ctgacatagc caagaaacca actgaaacga tttgtgcacc actcactgtc
      841 ttttttgatg gtagagttga tggtcaagta gacttattta gaaatgcccg taatggtgtt
      901 cttattacag aaggtagtgt taaaggttta caaccatctg taggtcccaa acaagctagt
      961 cttaatggag tcacattaat tggagaagcc gtaaaaacac agttcaatta ttataagaaa
     1021 gttgatggtg ttgtccaaca attacctgaa acttacttta ctcagagtag aaatttacaa
     1081 gaatttaaac ccaggagtca aatggaaatt gatttcttag aattagctat ggatgaattc
     1141 attgaacggt ataaattaga aggctatgcc ttcgaacata tcgtttatgg agattttagt
     1201 catagtcagt taggtggttt acatctactg attggactag ctaaacgttt taaggaatca
     1261 ccttttgaat tagaagattt tattcctatg gacagtacag ttaaaaacta tttcataaca
     1321 gatgcgcaaa caggttcatc taagtgtgtg tgttctgtta ttgatttatt acttgatgat
     1381 tttgttgaaa taataaaatc ccaagattta tctgtagttt ctaaggttgt caaagtgact
     1441 attgactata cagaaatttc atttatgctt tggtgtaaag atggccatgt agaaacattt
     1501 tacccaaaat tactcgtgcc gagaggttct gattataaag atcatgatgg tgattataaa
     1561 gatcatgata ttgattataa agatgatgat gataaataaa caggcccctt ttcctttgtc
     1621 gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac atccgctcta
     1681 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt tttttaatag
     1741 ttatgttagt attaagaacg ttatttatnn ttcaaatttt tctttttttt ctgtacaaag
     1801 cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt tttgattcgg
     1861 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
     1921 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
     1981 caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg
     2041 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa
     2101 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat
     2161 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg
     2221 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgagg
     2281 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca
     2341 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
     2401 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
     2461 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca
     2521 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa
     2581 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag
     2641 atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga tctgacatta
     2701 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt
     2761 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg
     2821 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
     2881 agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     2941 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     3001 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     3061 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     3121 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa
     3181 atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt
     3241 gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga
     3301 agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc
     3361 gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt
     3421 atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga
     3481 ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta
     3541 taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca
     3601 atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta
     3661 gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga
     3721 tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag
     3781 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca gagcgctttt
     3841 ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg aacttcccga
     3901 gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac tcactggctc
     3961 accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga gatccttttt
     4021 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4081 tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca gagcgcagat
     4141 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4201 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4261 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4321 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4381 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4441 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc caggggtaaa
     4501 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc atcgattttt
     4561 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac
     4621 ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac cgtcatcacc
     4681 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     4741 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     4801 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt accaatgctt
     4861 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4921 ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca gtgctgcaat
     4981 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5041 gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt ctattaattg
     5101 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5161 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5221 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5281 cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5341 agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg tgactggtga
     5401 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5461 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5521 acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5581 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5641 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5701 aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     5761 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa
     5821 gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt taaatggtca
     5881 gttaga
//
