LOCUS       Exported                5797 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 endoRNAse.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046358
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5797)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5797)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5797
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        19..426
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1475..1492
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1493..1516
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1561..1579)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1930..2733
                     /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      2816..3798
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3876..3905
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3927..4514
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4526..4553
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4577..4768)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4769..5629)
                     /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     5392..5411
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5630..5732)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5733..5764
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 tcagttagag cggccgcccc acacaccata gcttcaaaat gtttctactc cttttttact
       61 cttccagatt ttctcggact ccgcgcatcg ccgtaccact tcaaaacacc caagcacagc
      121 atactaaatt tcccctcttt cttcctctag ggtgtcgtta attacccgta ctaaaggttt
      181 ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt cgaaaaaggc aataaaaatt
      241 tttatcacgt ttctttttct tgaaaatttt tttttttgat ttttttctct ttcgatgacc
      301 tcccattgat atttaagtta ataaacggtc ttcaatttct caagtttcag tttcattttt
      361 cttgttctat tacaactttt tttacttctt gctcattaga aagaaagcat agcaatctaa
      421 tctaagtttt aattacaaaa tgttagaaaa tgtggctttt aatgttgtaa ataagggaca
      481 ctttgatgga caacagggtg aagtaccagt ttctatcatt aataacactg tttacacaaa
      541 agttgatggt gttgatgtag aattgtttga aaataaaaca acattacctg ttaatgtagc
      601 atttgagctt tgggctaagc gcaacattaa accagtacca gaggtgaaaa tactcaataa
      661 tttgggtgtg gacattgctg ctaatactgt gatctgggac tacaaaagag atgctccagc
      721 acatatatct actattggtg tttgttctat gactgacata gccaagaaac caactgaaac
      781 gatttgtgca ccactcactg tcttttttga tggtagagtt gatggtcaag tagacttatt
      841 tagaaatgcc cgtaatggtg ttcttattac agaaggtagt gttaaaggtt tacaaccatc
      901 tgtaggtccc aaacaagcta gtcttaatgg agtcacatta attggagaag ccgtaaaaac
      961 acagttcaat tattataaga aagttgatgg tgttgtccaa caattacctg aaacttactt
     1021 tactcagagt agaaatttac aagaatttaa acccaggagt caaatggaaa ttgatttctt
     1081 agaattagct atggatgaat tcattgaacg gtataaatta gaaggctatg ccttcgaaca
     1141 tatcgtttat ggagatttta gtcatagtca gttaggtggt ttacatctac tgattggact
     1201 agctaaacgt tttaaggaat caccttttga attagaagat tttattccta tggacagtac
     1261 agttaaaaac tatttcataa cagatgcgca aacaggttca tctaagtgtg tgtgttctgt
     1321 tattgattta ttacttgatg attttgttga aataataaaa tcccaagatt tatctgtagt
     1381 ttctaaggtt gtcaaagtga ctattgacta tacagaaatt tcatttatgc tttggtgtaa
     1441 agatggccat gtagaaacat tttacccaaa attactcgtg ccgagaggtt cttggagcca
     1501 tccgcagttt gaaaaataaa caggcccctt ttcctttgtc gatatcatgt aattagttat
     1561 gtcacgctta cattcacgcc ctcctcccac atccgctcta accgaaaagg aaggagttag
     1621 acaacctgaa gtctaggtcc ctatttattt tttttaatag ttatgttagt attaagaacg
     1681 ttatttatat ttcaaatttt tctttttttt ctgtacaaag cggccgccga accactagtg
     1741 cactgcagta cacggtttcc ttgaaatttt tttgattcgg taatctccga acagaaggaa
     1801 gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa
     1861 acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaacgtg caggaaacga
     1921 agataaatca tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt
     1981 gctgccaagc tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat
     2041 gttcgtacca ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta
     2101 ctaaaaacac atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta
     2161 aaggcattat ccgccaagta caatttttta ctcttcgagg acagaaaatt tgctgacatt
     2221 ggtaatacag tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac
     2281 attacgaatg cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca
     2341 gaagaagtaa caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc
     2401 tccctatcta ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat
     2461 tttgttatcg gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg
     2521 ttgattatga cacccggtgt gggtttagat gacaagggag atgcattggg tcaacagtat
     2581 agaaccgtgg atgatgtggt ttctacagga tctgacatta ttattgttgg aagaggacta
     2641 tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa
     2701 gcatatttga gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt
     2761 atactaaact cacaaattag agcttcaatt taattatatc agttattacc cgagtaacga
     2821 agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac
     2881 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca
     2941 acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt
     3001 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt
     3061 taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat
     3121 ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc
     3181 tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct
     3241 attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa
     3301 gctgcgggtg cattttttca agataaaggc atccccgatt atattctata ccgatgtgga
     3361 ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat
     3421 tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc
     3481 gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta
     3541 atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga
     3601 aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt
     3661 ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg
     3721 cgtttttggt tttttgaaag tgcgtcatca gagcgctttt ggttttcaaa agcgctctga
     3781 agttcctata ctttctagct agagaatagg aacttcccga gcggccgcgt gttacaagct
     3841 tgactccagc gtaactggac tgcaatcaac tcactggctc accttcacgg gtgggccttt
     3901 cttcggtaga aaatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3961 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4021 actctttttc cgaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4081 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4141 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4201 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4261 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4321 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4381 tcggaacagg agagcgcacg agggagcttc caggggtaaa cgcctggtat ctttatagtc
     4441 ctgtcgggtt tcgccacctc tgacttgagc atcgattttt gtgatgctcg tcaggggggc
     4501 ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac ccgaaggtga gccaggtgat
     4561 tacatttggg ccctcatcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     4621 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     4681 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     4741 taagggcggt tttttcctgt ttggtcactt accaatgctt aatcagtgag gcacctatct
     4801 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4861 cgatgcggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4921 caccggctcc agatttatca gcaataaacc agccagccgg gagggccgag cgcagaagtg
     4981 atcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5041 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5101 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5161 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgcca
     5221 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5281 ctgtcatgcc atccgtgaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5341 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5401 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgtaaac
     5461 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5521 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5581 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcatt ttagcttcct
     5641 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
     5701 ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa gcctccggtc ggaggctttt
     5761 gactttctgc tatggaggtc aggtatgatt taaatgg
//
