LOCUS       Exported                6306 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp4.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046208
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6306)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6306)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6306
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        46..487
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    46..163
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     458..481
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1996..2013
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2014..2037
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2082..2100)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2451..3254
                     /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      3337..4319
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4397..4426
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4448..5035
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      5047..5074
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5098..5289)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5290..6150)
                     /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     5913..5932
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6151..6253)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      6254..6285
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 tgatttaaat ggtcagttag agcggccgct tgctatattg aagtacggat tagaagccgc
       61 cgagcgggcg acagccctcc gacggaagac tctcctccgt gcgtcctcgt cttcaccggt
      121 cgcgttcctg aaacgcagat gtgcctcgcg ccgcactgct ccgaacaata aagattctac
      181 aatactagct tttatggtta tgaagaggaa aaattggcag taacctggcc ccacaaacct
      241 tcaaattaac gaatcaaatt aacaaccata ggatgataat gcgattagtt ttttagcctt
      301 atttctgggg taattaatca gcgaagcgat gatttttgat ctattaacag atatataaat
      361 ggaaaagctg cataaccact ttaactaata ctttcaacat tttcagtttg tattacttct
      421 tattcaaatg tcataaaagt atcaacaaaa aattgttaat atacctctat actttaacgt
      481 caaggagaaa aaactataat gattgttaat aattggttga agcagttaat taaagttaca
      541 cttgtgttcc tttttgttgc tgctattttc tatttaataa cacctgttca tgtcatgtct
      601 aaacatactg acttttcaag tgaaatcata ggatacaagg ctattgatgg tggtgtcact
      661 cgtgacatag catctacaga tacttgtttt gctaacaaac atgctgattt tgacacatgg
      721 tttagccagc gtggtggtag ttatactaat gacaaagctt gcccattgat tgctgcagtc
      781 ataacaagag aagtgggttt tgtcgtgcct ggtttgcctg gcacgatatt acgcacaact
      841 aatggtgact ttttgcattt cttacctaga gtttttagtg cagttggtaa catctgttac
      901 acaccatcaa aacttataga gtacactgac tttgcaacat cagcttgtgt tttggctgct
      961 gaatgtacaa tttttaaaga tgcttctggt aagccagtac catattgtta tgataccaat
     1021 gtactagaag gttctgttgc ttatgaaagt ttacgccctg acacacgtta tgtgctcatg
     1081 gatggctcta ttattcaatt tcctaacacc taccttgaag gttctgttag agtggtaaca
     1141 acttttgatt ctgagtactg taggcacggc acttgtgaaa gatcagaagc tggtgtttgt
     1201 gtatctacta gtggtagatg ggtacttaac aatgattatt acagatcttt accaggagtt
     1261 ttctgtggtg tagatgctgt aaatttactt actaatatgt ttacaccact aattcaacct
     1321 attggtgctt tggacatatc agcatctata gtagctggtg gtattgtagc tatcgtagta
     1381 acatgccttg cctactattt tatgaggttt agaagagctt ttggtgaata cagtcatgta
     1441 gttgccttta atactttact attccttatg tcattcactg tactctgttt aacaccagtt
     1501 tactcattct tacctggtgt ttattctgtt atttacttgt acttgacatt ttatcttact
     1561 aatgatgttt cttttttagc acatattcag tggatggtta tgttcacacc tttagtacct
     1621 ttctggataa caattgctta tatcatttgt atttccacaa agcatttcta ttggttcttt
     1681 agtaattacc taaagagacg tgtagtcttt aatggtgttt cctttagtac ttttgaagaa
     1741 gctgcgctgt gcaccttttt gttaaataaa gaaatgtatc taaagttgcg tagtgatgtg
     1801 ctattacctc ttacgcaata taatagatac ttagctcttt ataataagta caagtatttt
     1861 agtggagcaa tggatacaac tagctacaga gaagctgctt gttgtcatct cgcaaaggct
     1921 ctcaatgact tcagtaactc aggttctgat gttctttacc aaccaccaca aacctctatc
     1981 acctcagctg ttttgctcgt gccgagaggt tcttggagcc atccgcagtt tgaaaaataa
     2041 acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt acattcacgc
     2101 cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc
     2161 cctatttatt ttttttaata gttatgttag tattaagaac gttatttata tttcaaattt
     2221 ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt acacggtttc
     2281 cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca
     2341 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat
     2401 tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag
     2461 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata
     2521 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat
     2581 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata
     2641 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt
     2701 acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca gtcaaattgc
     2761 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg
     2821 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac
     2881 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat
     2941 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg
     3001 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg
     3061 tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg
     3121 tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg
     3181 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg
     3241 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta
     3301 gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt gcttcatttt
     3361 gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     3421 tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca
     3481 tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     3541 gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa agaatctata
     3601 cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac aaagcatctt
     3661 agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa ctttttgcac
     3721 tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct tccataaaaa
     3781 aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt gcattttttc
     3841 aagataaagg catccccgat tatattctat accgatgtgg attgcgcata ctttgtgaac
     3901 agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg tttcttctat
     3961 tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt tcgattcact
     4021 ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag ataaacataa
     4081 aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat gggtaggtta
     4141 tatagggata tagcacagag atatatagca aagagatact tttgagcaat gtttgtggaa
     4201 gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa
     4261 gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat actttctagc
     4321 tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag cgtaactgga
     4381 ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag aaaatcaaag
     4441 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4501 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaggtaac
     4561 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4621 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4681 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4741 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4801 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4861 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4921 gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4981 ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5041 cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg gccctcatca
     5101 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     5161 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     5221 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     5281 tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5341 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg agggcttacc
     5401 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5461 agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa ctttatccgc
     5521 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5581 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5641 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5701 caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt
     5761 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtgag
     5821 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5881 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5941 aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct
     6001 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6061 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6121 aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg aaaatctcga
     6181 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     6241 tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg ctatggaggt
     6301 caggta
//
