LOCUS       Exported                5388 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 leader protein.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046541
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5388)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5388)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5388
                     /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..1098
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1143..1161)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1512..2315
                     /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      2398..3380
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3458..3487
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3509..4096
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4108..4135
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4159..4350)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4351..5211)
                     /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     4974..4993
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5212..5314)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5315..5346
                     /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 ctgattataa agatcatgat ggtgattata aagatcatga tattgattat
     1081 aaagatgatg atgataaata aacaggcccc ttttcctttg tcgatatcat gtaattagtt
     1141 atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa ggaaggagtt
     1201 agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta gtattaagaa
     1261 cgttatttat atttcaaatt tttctttttt ttctgtacaa agcggccgcc gaaccactag
     1321 tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc gaacagaagg
     1381 aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag
     1441 aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg tgcaggaaac
     1501 gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg
     1561 ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg
     1621 atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt
     1681 tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc
     1741 taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa tttgctgaca
     1801 ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag
     1861 acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg
     1921 cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg
     1981 gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag
     2041 attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt
     2101 ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg ggtcaacagt
     2161 atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt ggaagaggac
     2221 tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg
     2281 aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat
     2341 gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta cccgagtaac
     2401 gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa
     2461 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc gctattttac
     2521 caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga gcgctaattt
     2581 ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg agagcgctat
     2641 tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc cgagagcgct
     2701 atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc tataatgcag
     2761 tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct actttggtgt
     2821 ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg attactagcg
     2881 aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta taccgatgtg
     2941 gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat tggtcagaaa
     3001 attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat gtttacattt
     3061 tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt gtctaaagag
     3121 taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc
     3181 gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc aaagagatac
     3241 ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt tacagtccgg
     3301 tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt ttggttttca aaagcgctct
     3361 gaagttccta tactttctag ctagagaata ggaacttccc gagcggccgc gtgttacaag
     3421 cttgactcca gcgtaactgg actgcaatca actcactggc tcaccttcac gggtgggcct
     3481 ttcttcggta gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3541 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3601 caactctttt tccgaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3661 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3721 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3781 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3841 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3901 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3961 ggtcggaaca ggagagcgca cgagggagct tccaggggta aacgcctggt atctttatag
     4021 tcctgtcggg tttcgccacc tctgacttga gcatcgattt ttgtgatgct cgtcaggggg
     4081 gcggagccta tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt gagccaggtg
     4141 attacatttg ggccctcatc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     4201 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     4261 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     4321 gttaagggcg gttttttcct gtttggtcac ttaccaatgc ttaatcagtg aggcacctat
     4381 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4441 tacgatgcgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4501 ctcaccggct ccagatttat cagcaataaa ccagccagcc gggagggccg agcgcagaag
     4561 tgatcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4621 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4681 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4741 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgc
     4801 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4861 tactgtcatg ccatccgtga gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4921 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4981 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgtaa
     5041 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5101 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5161 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca ttttagcttc
     5221 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     5281 atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg tcggaggctt
     5341 ttgactttct gctatggagg tcaggtatga tttaaatggt cagttaga
//
