LOCUS       Exported                5346 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 leader protein.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046215
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5346)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5346)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5346
                     /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             1015..1032
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1033..1056
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1101..1119)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1470..2273
                     /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      2356..3338
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3416..3445
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3467..4054
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4066..4093
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4117..4308)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4309..5169)
                     /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     4932..4951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5170..5272)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5273..5304
                     /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 gagagccttg tccctggttt caacgagaaa acacacgtcc aactcagttt gcctgtttta
      541 caggttcgcg acgtgctcgt acgtggcttt ggagactccg tggaggaggt cttatcagag
      601 gcacgtcaac atcttaaaga tggcacttgt ggcttagtag aagttgaaaa aggcgttttg
      661 cctcaacttg aacagcccta tgtgttcatc aaacgttcgg atgctcgaac tgcacctcat
      721 ggtcatgtta tggttgagct ggtagcagaa ctcgaaggca ttcagtacgg tcgtagtggt
      781 gagacacttg gtgtccttgt ccctcatgtg ggcgaaatac cagtggctta ccgcaaggtt
      841 cttcttcgta agaacggtaa taaaggagct ggtggccata gttacggcgc cgatctaaag
      901 tcatttgact taggcgacga gcttggcact gatccttatg aagattttca agaaaactgg
      961 aacactaaac atagcagtgg tgttacccgt gaactcatgc gtgagcttaa cggactcgtg
     1021 ccgagaggtt cttggagcca tccgcagttt gaaaaataaa caggcccctt ttcctttgtc
     1081 gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac atccgctcta
     1141 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt tttttaatag
     1201 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacaaag
     1261 cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt tttgattcgg
     1321 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
     1381 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
     1441 caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg
     1501 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa
     1561 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat
     1621 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg
     1681 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgagg
     1741 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca
     1801 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
     1861 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
     1921 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca
     1981 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa
     2041 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag
     2101 atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga tctgacatta
     2161 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt
     2221 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg
     2281 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
     2341 agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     2401 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     2461 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     2521 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     2581 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa
     2641 atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt
     2701 gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga
     2761 agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc
     2821 gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt
     2881 atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga
     2941 ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta
     3001 taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca
     3061 atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta
     3121 gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga
     3181 tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag
     3241 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca gagcgctttt
     3301 ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg aacttcccga
     3361 gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac tcactggctc
     3421 accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga gatccttttt
     3481 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3541 tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca gagcgcagat
     3601 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3661 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3721 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3781 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3841 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3901 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc caggggtaaa
     3961 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc atcgattttt
     4021 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac
     4081 ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac cgtcatcacc
     4141 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     4201 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     4261 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt accaatgctt
     4321 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4381 ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca gtgctgcaat
     4441 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4501 gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt ctattaattg
     4561 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4621 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4681 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4741 cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4801 agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg tgactggtga
     4861 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4921 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4981 acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5041 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5101 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5161 aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     5221 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa
     5281 gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt taaatggtca
     5341 gttaga
//
