LOCUS       sequence-297985-        4987 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 E (envelope).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4987)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4987)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046261
FEATURES             Location/Qualifiers
     source          1..4987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             656..673
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             674..697
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(742..760)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1111..1914
                     /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      1997..2979
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3057..3086
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3108..3695
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3707..3734)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3758..3949)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3950..4741,4742..4810))
                     /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     4573..4592
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4811..4913)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4914..4945)
                     /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 ttnnnnnnnt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atgtattctt ttgttagcga ggaaacggga actcttatag taaactcggt
      481 gctattattc ctggctttcg tcgtttttct cttggttacc ttagcaattt tgactgcctt
      541 gaggttgtgt gcgtactgct gtaatatcgt caacgtttca ttggtcaagc cttctttcta
      601 cgtttactcc agagtcaaaa atttgaactc ctctcgcgtc ccagatttat tggttctcgt
      661 gccgagaggt tcttggagcc atccgcagtt tgaaaaataa acaggcccct tttcctttgt
      721 cgatatcatg taattagtta tgtcacgctt acattcacgc cctcctccca catccgctct
      781 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt ttttttaata
      841 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaaa
      901 gcggccgccg aaccactagt gcactgcagt acacggtttc cttgaaattt ttttgattcg
      961 gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata
     1021 cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga
     1081 acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct
     1141 gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca
     1201 aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca
     1261 ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg
     1321 gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgag
     1381 gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac
     1441 agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt
     1501 agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta
     1561 gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac
     1621 attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga
     1681 agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga
     1741 gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg tttctacagg atctgacatt
     1801 attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt
     1861 tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact
     1921 gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat
     1981 cagttattac ccgagtaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc
     2041 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     2101 cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc
     2161 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     2221 tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa
     2281 aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt
     2341 tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag
     2401 aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg
     2461 cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat
     2521 tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg
     2581 attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt
     2641 ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac
     2701 aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt
     2761 agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag
     2821 atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta
     2881 gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcatc agagcgcttt
     2941 tggttttcaa aagcgctctg aagttcctat actttctagc tagagaatag gaacttcccg
     3001 agcggccgcg tgttacaagc ttgactccag cgtaactgga ctgcaatcaa ctcactggct
     3061 caccttcacg ggtgggcctt tcttcggtag aaaatcaaag gatcttcttg agatcctttt
     3121 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3181 ttgccggatc aagagctacc aactcttttt ccgaggtaac tggcttcagc agagcgcaga
     3241 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3301 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3361 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3421 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3481 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3541 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccaggggtaa
     3601 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag crtcgatttt
     3661 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgca gaaaggccca
     3721 cccgaaggtg agccaggtga ttacatttgg gccctcatca gaggttttca ccgtcatcac
     3781 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     3841 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     3901 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact taccaatgct
     3961 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4021 tccccgtcgt gtagataact acgatgcggg agggcttacc atctggcccc agtgctgcaa
     4081 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4141 ggagggccga gcgcagaagt gatcctgcaa ctttatccgc ctccatccag tctattaatt
     4201 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4261 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4321 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4381 tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4441 cagcactgca taattctctt actgtcatgc catccgtgag atgcttttct gtgactggtg
     4501 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4561 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4621 aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4681 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4741 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4801 gaatactcat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     4861 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa
     4921 agcctccggt cggaggcttt tgactttctg ctatggaggt caggtatgat ttaaatggtc
     4981 agttaga
//
