LOCUS       Exported                6726 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp2.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046430
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6726)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6726)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6726
                     /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             2395..2412
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2413..2436
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2481..2499)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2850..3653
                     /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      3736..4718
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4796..4825
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4847..5434
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      5446..5473
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5497..5688)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5689..6549)
                     /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     6312..6331
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6550..6652)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      6653..6684
                     /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 gcatacacga ggtatgtaga caataacttt tgcggtcctg atggataccc actggaatgt
      541 atcaaagatt tgttagctcg tgccggtaag gctagctgta ccctttctga gcaattggac
      601 ttcatagaca ctaagagagg tgtttactgt tgtcgcgaac atgaacacga aattgcctgg
      661 tataccgaga gatcagaaaa atcctacgaa ttacagactc cattcgaaat taagttggct
      721 aagaagtttg atacattcaa cggcgaatgc cccaatttcg tctttccatt gaactctatc
      781 attaaaacta tccaaccgag agtcgagaag aagaaattgg atggttttat gggtagaatt
      841 agatcggtgt acccagttgc ttccccaaat gaatgtaacc aaatgtgtct atctactttg
      901 atgaagtgtg accactgtgg tgaaaccagt tggcaaacag gcgacttcgt taaggctaca
      961 tgtgaatttt gcggtaccga gaacctcact aaggaaggtg cgactacttg tggttattta
     1021 cctcaaaatg cagttgtcaa gatttactgt ccagcttgcc ataactctga agttggacct
     1081 gaacacagct tggctgaata ccacaacgaa tccggtttaa agaccatcct tcgaaaaggt
     1141 ggtcgtacca tagccttcgg gggttgcgtc ttctcatatg ttggttgtca taataagtgt
     1201 gcctattggg ttccaagagc ttctgctaat atcggatgta accacacggg tgtggtcggt
     1261 gaaggctctg agggtttgaa cgataacttg ttggaaatac ttcagaagga aaaggtcaac
     1321 attaacatcg ttggtgactt taaattgaat gaagaaattg ctattatcct agcgtccttc
     1381 tcggcctcaa cttccgcttt tgtagaaact gttaagggtc ttgattacaa agctttcaag
     1441 caaatcgtag aatcatgcgg taatttcaag gttaccaagg ggaaagccaa aaaaggtgcc
     1501 tggaacattg gtgaacaaaa gtctatactg agtccattat atgctttcgc atctgaagct
     1561 gctagagtcg tcagatctat tttttccaga actctcgaaa ccgctcaaaa ttcagtgaga
     1621 gttttgcaaa aagcagccat tactatcttg gacggtattt cccagtactc tttgaggtta
     1681 atcgatgcta tgatgtttac ctccgacttg gctactaata acctggtcgt catggcttac
     1741 ataaccggtg gcgttgttca attgacatct caatggctaa ctaatatatt cggtactgtt
     1801 tatgaaaagc tcaagccagt cttggattgg ttggaggaaa agttcaagga gggcgttgag
     1861 tttttacggg atggttggga aatcgttaag ttcatttcta catgtgcctg tgaaattgtc
     1921 ggtggtcaaa tcgtgacctg tgctaaggaa attaaggagt cggttcaaac tttctttaag
     1981 ttggtgaaca aatttttagc tctctgtgct gattcgatta tcatcggagg agcgaaattg
     2041 aaggcactga acttgggtga aaccttcgtc acccactcca aaggtctgta ccgtaaatgt
     2101 gttaaatcaa gagaagaaac tggtttgcta atgcctttga aggccccgaa ggaaattatc
     2161 ttcttggaag gtgaaacatt accaacagag gttttaacgg aagaggtagt tttgaagacg
     2221 ggggatttgc agccgttaga acaaccaacc tctgaagccg tggaagctcc cctagttgga
     2281 acgccagttt gcattaacgg tctgatgtta ttggaaatca aggacaccga aaagtactgt
     2341 gctctcgctc caaacatgat ggtcactaac aataccttca ctttgaaggg tggtctcgtg
     2401 ccgagaggtt cttggagcca tccgcagttt gaaaaataaa caggcccctt ttcctttgtc
     2461 gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac atccgctcta
     2521 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt tttttaatag
     2581 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacaaag
     2641 cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt tttgattcgg
     2701 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
     2761 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
     2821 caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg
     2881 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa
     2941 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat
     3001 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg
     3061 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgagg
     3121 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca
     3181 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
     3241 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
     3301 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca
     3361 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa
     3421 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag
     3481 atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga tctgacatta
     3541 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt
     3601 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg
     3661 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
     3721 agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     3781 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     3841 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     3901 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     3961 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa
     4021 atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt
     4081 gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga
     4141 agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc
     4201 gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt
     4261 atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga
     4321 ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta
     4381 taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca
     4441 atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta
     4501 gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga
     4561 tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag
     4621 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca gagcgctttt
     4681 ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg aacttcccga
     4741 gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac tcactggctc
     4801 accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga gatccttttt
     4861 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4921 tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca gagcgcagat
     4981 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5041 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5101 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5161 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5221 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5281 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc caggggtaaa
     5341 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc atcgattttt
     5401 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac
     5461 ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac cgtcatcacc
     5521 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     5581 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     5641 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt accaatgctt
     5701 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5761 ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca gtgctgcaat
     5821 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5881 gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt ctattaattg
     5941 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6001 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6061 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6121 cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6181 agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg tgactggtga
     6241 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6301 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6361 acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6421 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6481 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6541 aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     6601 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa
     6661 gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt taaatggtca
     6721 gttaga
//
