LOCUS       Exported                5187 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp9.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046550
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5187)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5187)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5187
                     /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             814..831
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             832..897
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(942..960)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1311..2114
                     /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      2197..3179
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3257..3286
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3308..3895
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      3907..3934
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3958..4149)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4150..5010)
                     /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     4773..4792
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5011..5113)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5114..5145
                     /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 aatgagctta gtcctgttgc actacgacag atgtcttgtg ctgccggtac tacacaaact
      541 gcttgcactg atgacaatgc gttagcttac tacaacacaa caaagggagg taggtttgta
      601 cttgcactgt tatccgattt acaggatttg aaatgggcta gattccctaa gagtgatgga
      661 actggtacta tctatacaga actggaacca ccttgtaggt ttgttacaga cacacctaaa
      721 ggtcctaaag tgaagtattt atactttatt aaaggattaa acaacctaaa tagaggtatg
      781 gtacttggta gtttagctgc cacagtacgt ctactcgtgc cgagaggttc tgattataaa
      841 gatcatgatg gtgattataa agatcatgat attgattata aagatgatga tgataaataa
      901 acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc
      961 cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
     1021 cctatttatt ttttttaata gttatgttag tattaagaac gttatttata tttcaaattt
     1081 ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt acacggtttc
     1141 cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
     1201 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
     1261 tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag
     1321 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
     1381 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
     1441 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
     1501 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
     1561 acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca gtcaaattgc
     1621 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
     1681 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
     1741 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
     1801 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
     1861 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     1921 tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     1981 tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     2041 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     2101 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     2161 gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt gcttcatttt
     2221 gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     2281 tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca
     2341 tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     2401 gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata
     2461 cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt
     2521 agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac
     2581 tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa
     2641 aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc
     2701 aagataaagg catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac
     2761 agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat
     2821 tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact
     2881 ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa
     2941 aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta
     3001 tatagggata tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa
     3061 gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa
     3121 gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctagc
     3181 tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag cgtaactgga
     3241 ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag aaaatcaaag
     3301 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3361 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaggtaac
     3421 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3481 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3541 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3601 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3661 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3721 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3781 gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3841 ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3901 cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg gccctcatca
     3961 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     4021 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     4081 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     4141 tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4201 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg agggcttacc
     4261 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4321 agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa ctttatccgc
     4381 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4441 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4501 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4561 caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt
     4621 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtgag
     4681 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4741 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4801 aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct
     4861 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4921 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4981 aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg aaaatctcga
     5041 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     5101 tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg ctatggaggt
     5161 caggtatgat ttaaatggtc agttaga
//
