LOCUS       Exported                7333 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp4.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046239
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7333)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7333)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7333
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1928..1945
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(3088..3106)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3457..4260
                     /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      4343..5325
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5403..5432
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5454..6041
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      6053..6080
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6104..6295)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6296..7156)
                     /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     6919..6938
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7157..7259)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      7260..7291
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atgattgtta ataattggtt gaagcagtta attaaagtta cacttgtgtt
      481 cctttttgtt gctgctattt tctatttaat aacacctgtt catgtcatgt ctaaacatac
      541 tgacttttca agtgaaatca taggatacaa ggctattgat ggtggtgtca ctcgtgacat
      601 agcatctaca gatacttgtt ttgctaacaa acatgctgat tttgacacat ggtttagcca
      661 gcgtggtggt agttatacta atgacaaagc ttgcccattg attgctgcag tcataacaag
      721 agaagtgggt tttgtcgtgc ctggtttgcc tggcacgata ttacgcacaa ctaatggtga
      781 ctttttgcat ttcttaccta gagtttttag tgcagttggt aacatctgtt acacaccatc
      841 aaaacttata gagtacactg actttgcaac atcagcttgt gttttggctg ctgaatgtac
      901 aatttttaaa gatgcttctg gtaagccagt accatattgt tatgatacca atgtactaga
      961 aggttctgtt gcttatgaaa gtttacgccc tgacacacgt tatgtgctca tggatggctc
     1021 tattattcaa tttcctaaca cctaccttga aggttctgtt agagtggtaa caacttttga
     1081 ttctgagtac tgtaggcacg gcacttgtga aagatcagaa gctggtgttt gtgtatctac
     1141 tagtggtaga tgggtactta acaatgatta ttacagatct ttaccaggag ttttctgtgg
     1201 tgtagatgct gtaaatttac ttactaatat gtttacacca ctaattcaac ctattggtgc
     1261 tttggacata tcagcatcta tagtagctgg tggtattgta gctatcgtag taacatgcct
     1321 tgcctactat tttatgaggt ttagaagagc ttttggtgaa tacagtcatg tagttgcctt
     1381 taatacttta ctattcctta tgtcattcac tgtactctgt ttaacaccag tttactcatt
     1441 cttacctggt gtttattctg ttatttactt gtacttgaca ttttatctta ctaatgatgt
     1501 ttctttttta gcacatattc agtggatggt tatgttcaca cctttagtac ctttctggat
     1561 aacaattgct tatatcattt gtatttccac aaagcatttc tattggttct ttagtaatta
     1621 cctaaagaga cgtgtagtct ttaatggtgt ttcctttagt acttttgaag aagctgcgct
     1681 gtgcaccttt ttgttaaata aagaaatgta tctaaagttg cgtagtgatg tgctattacc
     1741 tcttacgcaa tataatagat acttagctct ttataataag tacaagtatt ttagtggagc
     1801 aatggataca actagctaca gagaagctgc ttgttgtcat ctcgcaaagg ctctcaatga
     1861 cttcagtaac tcaggttctg atgttcttta ccaaccacca caaacctcta tcacctcagc
     1921 tgttttgctc gtgccgagag gttctaaaat cgaggaaggt aagctagtca tttggataaa
     1981 tggagataag ggttacaacg gtttggcaga agtaggcaag aaatttgaaa aggacacagg
     2041 tatcaaggtg acggttgaac atccagacaa gttagaagaa aaattccctc aagttgctgc
     2101 tactggtgat ggtcccgaca ttattttctg ggcccacgat cgttttggtg gatatgctca
     2161 gtcaggtttg ctggctgaga tcaccccaga caaggccttc caagataaat tgtacccatt
     2221 cacttgggat gctgtcagat acaacggtaa gcttatagca tatccaattg ctgttgaagc
     2281 cttatcgttg atctacaata aggacttgtt acctaaccca ccaaagacat gggaagaaat
     2341 tccagctttg gataaagagc taaaggccaa gggaaaaagc gctttgatgt ttaacttaca
     2401 agaaccttat ttcacctggc cattaatcgc tgctgacggt ggttacgcat ttaaatacga
     2461 aaacggtaag tatgacatta aggatgtcgg cgttgataat gctggtgcaa aggctggttt
     2521 gaccttcttg gttgatttaa taaagaataa acacatgaac gctgacactg actactctat
     2581 cgccgaagcc gctttcaaca agggtgaaac tgctatgacc ataaatggtc catgggcctg
     2641 gagtaacatt gatacttcta aagtaaacta tggtgtcact gttttgccta cattcaaagg
     2701 ccaaccatcc aagccctttg tcggtgttct atccgctggc atcaatgctg cttctccaaa
     2761 caaggagcta gctaaagaat ttttggaaaa ttacttgtta actgacgaag gtttggaggc
     2821 cgttaacaag gacaagccat tgggtgctgt cgctttgaaa tcttacgaag aagaattggc
     2881 taaggatccc agaattgccg ctaccatgga gaacgcccaa aagggtgaaa ttatgccaaa
     2941 catcccacaa atgtcagcct tctggtacgc agttaggact gccgtgatta atgctgcttc
     3001 cggtagacaa accgtcgacg aagcattaaa agatgctcaa acgtaaacag gccccttttc
     3061 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc
     3121 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     3181 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     3241 tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt
     3301 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
     3361 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
     3421 cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa
     3481 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag
     3541 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt
     3601 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt
     3661 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc
     3721 ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt
     3781 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt
     3841 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg
     3901 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact
     3961 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg
     4021 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac
     4081 aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct
     4141 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt
     4201 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
     4261 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
     4321 ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg
     4381 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     4441 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     4501 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     4561 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     4621 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     4681 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     4741 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     4801 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     4861 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     4921 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     4981 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     5041 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     5101 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     5161 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     5221 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag
     5281 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
     5341 ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca
     5401 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat
     5461 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5521 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag
     5581 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5641 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5701 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5761 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5821 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5881 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5941 gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc
     6001 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa
     6061 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt
     6121 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     6181 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     6241 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc
     6301 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6361 cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg
     6421 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6481 cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta
     6541 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6601 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6661 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6721 gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg
     6781 ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga
     6841 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6901 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6961 ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt
     7021 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7081 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7141 aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     7201 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     7261 gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa
     7321 atggtcagtt aga
//
