LOCUS       Exported                7467 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 3'-to-5' exonuclease.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046252
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7467)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7467)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7467
                     /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             2062..2079
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(3222..3240)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3591..4394
                     /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      4477..5459
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5537..5566
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5588..6175
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      6187..6214
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6238..6429)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6430..7290)
                     /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     7053..7072
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7291..7393)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      7394..7425
                     /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 gccgagaatg tcacaggcct attcaaggat tgctcgaaag tgatcaccgg tttacaccct
      541 acgcaggctc caactcattt gtccgttgac accaagttta agactgaagg attgtgtgtt
      601 gacataccag gtattcccaa agatatgact taccgtaggc tgatttctat gatgggtttc
      661 aagatgaact atcaagttaa cggttaccca aatatgttta tcaccagaga agaagctatt
      721 agacacgtac gtgcatggat cggtttcgat gtcgaaggtt gtcatgctac tagagaagcg
      781 gtcggtacaa acttgccatt acaacttggc ttttcaaccg gtgttaattt ggttgctgtt
      841 cctacagggt atgtcgacac tccaaacaac acggatttct ctcgagtgtc cgccaagcca
      901 ccacctggtg atcaattcaa acacttgatc ccgctcatgt acaagggttt gccttggaac
      961 gttgtcagaa ttaaaatagt acaaatgtta tctgatactt tgaagaatct gagtgacaga
     1021 gttgtcttcg tcctttgggc tcacggtttt gagctaacca gcatgaagta ctttgtgaag
     1081 attggaccag aacggacttg ttgcttgtgt gacagaagag ccacttgttt ctccaccgct
     1141 agcgatacat atgcatgttg gcatcattct attggtttcg actacgttta caaccccttt
     1201 atgatcgacg ttcaacagtg gggtttcacc ggtaatttgc aatcgaacca cgacttgtat
     1261 tgtcaagttc acggcaatgc tcacgtggct tcatgcgatg ctattatgac aaggtgttta
     1321 gccgtccatg aatgtttcgt taaaagagta gactggacta tcgaataccc aattatcggt
     1381 gatgaactca agattaacgc tgcctgtaga aaggtccaac atatggttgt aaaggcagct
     1441 ctattagctg acaaatttcc agttttgcac gatataggga atccaaaggc cataaagtgt
     1501 gttccacaag ctgacgtcga atggaaattc tacgatgcgc aaccctgctc tgataaagct
     1561 tataagatcg aggaattgtt ttactcttac gctacccact ccgataagtt cactgacggt
     1621 gtgtgtcttt tctggaactg caatgttgat agatatccag caaactccat cgtttgtaga
     1681 ttcgacacgc gagtcttgag caacttgaac ttaccaggtt gtgatggtgg ttccttgtac
     1741 gtcaataaac atgcatttca cactcccgct ttcgacaagt ctgcttttgt taatttaaag
     1801 caattgcctt ttttctatta ctctgatagt ccatgcgaat ctcatggtaa gcaagtcgtt
     1861 tccgacattg attacgtacc actgaagtca gccacctgta taacaagatg taaccttgga
     1921 ggtgctgtct gccgccacca tgctaacgag tacagactat acttggatgc ctataacatg
     1981 atgatttcag ccggtttctc tttatgggtt tacaagcagt ttgacactta caacttgtgg
     2041 aacaccttca cgcgtttgca actcgtgccg agaggttcta aaatcgagga aggtaagcta
     2101 gtcatttgga taaatggaga taagggttac aacggtttgg cagaagtagg caagaaattt
     2161 gaaaaggaca caggtatcaa ggtgacggtt gaacatccag acaagttaga agaaaaattc
     2221 cctcaagttg ctgctactgg tgatggtccc gacattattt tctgggccca cgatcgtttt
     2281 ggtggatatg ctcagtcagg tttgctggct gagatcaccc cagacaaggc cttccaagat
     2341 aaattgtacc cattcacttg ggatgctgtc agatacaacg gtaagcttat agcatatcca
     2401 attgctgttg aagccttatc gttgatctac aataaggact tgttacctaa cccaccaaag
     2461 acatgggaag aaattccagc tttggataaa gagctaaagg ccaagggaaa aagcgctttg
     2521 atgtttaact tacaagaacc ttatttcacc tggccattaa tcgctgctga cggtggttac
     2581 gcatttaaat acgaaaacgg taagtatgac attaaggatg tcggcgttga taatgctggt
     2641 gcaaaggctg gtttgacctt cttggttgat ttaataaaga ataaacacat gaacgctgac
     2701 actgactact ctatcgccga agccgctttc aacaagggtg aaactgctat gaccataaat
     2761 ggtccatggg cctggagtaa cattgatact tctaaagtaa actatggtgt cactgttttg
     2821 cctacattca aaggccaacc atccaagccc tttgtcggtg ttctatccgc tggcatcaat
     2881 gctgcttctc caaacaagga gctagctaaa gaatttttgg aaaattactt gttaactgac
     2941 gaaggtttgg aggccgttaa caaggacaag ccattgggtg ctgtcgcttt gaaatcttac
     3001 gaagaagaat tggctaagga tcccagaatt gccgctacca tggagaacgc ccaaaagggt
     3061 gaaattatgc caaacatccc acaaatgtca gccttctggt acgcagttag gactgccgtg
     3121 attaatgctg cttccggtag acaaaccgtc gacgaagcat taaaagatgc tcaaacgtaa
     3181 acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc
     3241 cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
     3301 cctatttatt ttttttaata gttatgttag tattaagaac gttatttata tttcaaattt
     3361 ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt acacggtttc
     3421 cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
     3481 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
     3541 tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag
     3601 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
     3661 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
     3721 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
     3781 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
     3841 acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca gtcaaattgc
     3901 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
     3961 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
     4021 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
     4081 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
     4141 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     4201 tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     4261 tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     4321 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     4381 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     4441 gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt gcttcatttt
     4501 gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     4561 tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca
     4621 tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     4681 gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata
     4741 cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt
     4801 agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac
     4861 tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa
     4921 aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc
     4981 aagataaagg catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac
     5041 agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat
     5101 tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact
     5161 ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa
     5221 aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta
     5281 tatagggata tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa
     5341 gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa
     5401 gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctagc
     5461 tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag cgtaactgga
     5521 ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag aaaatcaaag
     5581 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5641 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaggtaac
     5701 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5761 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5821 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5881 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5941 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6001 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6061 gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6121 ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6181 cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg gccctcatca
     6241 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     6301 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     6361 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     6421 tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6481 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg agggcttacc
     6541 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6601 agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa ctttatccgc
     6661 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6721 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6781 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6841 caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt
     6901 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtgag
     6961 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7021 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7081 aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct
     7141 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7201 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7261 aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg aaaatctcga
     7321 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     7381 tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg ctatggaggt
     7441 caggtatgat ttaaatggtc agttaga
//
