LOCUS       Exported                5838 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 endoRNAse.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046393
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5838)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5838)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5838
                     /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..1548
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1593..1611)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1962..2765
                     /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      2848..3830
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3908..3937
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3959..4546
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4558..4585
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4609..4800)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4801..5661)
                     /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     5424..5443
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5662..5764)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5765..5796
                     /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 caccatcatc accatcatta aacaggcccc
     1561 ttttcctttg tcgatatcat gtaattagtt atgtcacgct tacattcacg ccctcctccc
     1621 acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat
     1681 tttttttaat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
     1741 ttctgtacaa agcggccgcc gaaccactag tgcactgcag tacacggttt ccttgaaatt
     1801 tttttgattc ggtaatctcc gaacagaagg aagaacgaag gaaggagcac agacttagat
     1861 tggtatatat acgcatatgt agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc
     1921 aactgcacag aacaaaaacg tgcaggaaac gaagataaat catgtcgaaa gctacatata
     1981 aggaacgtgc tgctactcat cctagtcctg ttgctgccaa gctatttaat atcatgcacg
     2041 aaaagcaaac aaacttgtgt gcttcattgg atgttcgtac caccaaggaa ttactggagt
     2101 tagttgaagc attaggtccc aaaatttgtt tactaaaaac acatgtggat atcttgactg
     2161 atttttccat ggagggcaca gttaagccgc taaaggcatt atccgccaag tacaattttt
     2221 tactcttcga ggacagaaaa tttgctgaca ttggtaatac agtcaaattg cagtactctg
     2281 cgggtgtata cagaatagca gaatgggcag acattacgaa tgcacacggt gtggtgggcc
     2341 caggtattgt tagcggtttg aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc
     2401 ttttgatgtt agcagaattg tcatgcaagg gctccctatc tactggagaa tatactaagg
     2461 gtactgttga cattgcgaag agcgacaaag attttgttat cggctttatt gctcaaagag
     2521 acatgggtgg aagagatgaa ggttacgatt ggttgattat gacacccggt gtgggtttag
     2581 atgacaaggg agatgcattg ggtcaacagt atagaaccgt ggatgatgtg gtttctacag
     2641 gatctgacat tattattgtt ggaagaggac tatttgcaaa gggaagggat gctaaggtag
     2701 agggtgaacg ttacagaaaa gcaggctggg aagcatattt gagaagatgc ggccagcaaa
     2761 actaaaaaac tgtattataa gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa
     2821 tttaattata tcagttatta cccgagtaac gaagcatctg tgcttcattt tgtagaacaa
     2881 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     2941 cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc atttttgtaa
     3001 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     3061 cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat acttcttttt
     3121 tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct tagattactt
     3181 tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca ctgtaggtcc
     3241 gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa aaagcctgac
     3301 tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt caagataaag
     3361 gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa cagaaagtga
     3421 tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta ttttgtctct
     3481 atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac tctatgaata
     3541 gttcttacta caattttttt gtctaaagag taatactaga gataaacata aaaaatgtag
     3601 aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt atatagggat
     3661 atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga agcggtattc
     3721 gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa agtgcgtcat
     3781 cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag ctagagaata
     3841 ggaacttccc gagcggccgc gtgttacaag cttgactcca gcgtaactgg actgcaatca
     3901 actcactggc tcaccttcac gggtgggcct ttcttcggta gaaaatcaaa ggatcttctt
     3961 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4021 cggtggtttg tttgccggat caagagctac caactctttt tccgaggtaa ctggcttcag
     4081 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4141 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4201 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4261 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4321 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4381 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4441 tccaggggta aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4501 gcatcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4561 agaaaggccc acccgaaggt gagccaggtg attacatttg ggccctcatc agaggttttc
     4621 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     4681 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     4741 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     4801 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4861 agttgcctga ctccccgtcg tgtagataac tacgatgcgg gagggcttac catctggccc
     4921 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4981 ccagccagcc gggagggccg agcgcagaag tgatcctgca actttatccg cctccatcca
     5041 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5101 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5161 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5221 ggttagctcc ttcggtcctc cgatcgttgc cagaagtaag ttggccgcag tgttatcact
     5281 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtga gatgcttttc
     5341 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5401 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5461 catcattgga aaacgttctt cggggcgtaa actctcaagg atcttaccgc tgttgagatc
     5521 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5581 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5641 acggaaatgt tgaatactca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     5701 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
     5761 atcaagtcaa aagcctccgg tcggaggctt ttgactttct gctatggagg tcaggtatga
     5821 tttaaatggt cagttaga
//
