LOCUS       Exported                5628 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF3a protein.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046437
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5628)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5628)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5628
                     /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             1303..1320
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1321..1338
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1383..1401)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1752..2555
                     /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      2638..3620
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3698..3727
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3749..4336
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4348..4375
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4399..4590)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4591..5451)
                     /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     5214..5233
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5452..5554)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5555..5586
                     /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 gacttattta tgagaatatt cacgatcggc acagtgaccc taaagcaggg tgagattaaa
      541 gatgctactc catcagattt cgttagggca accgctacta tccccattca agccagcttg
      601 ccttttggtt ggttgattgt aggagtcgct ctcttggctg ttttccaatc cgccagtaag
      661 atcattactt taaaaaagag atggcaattg gcgctttcta agggtgttca cttcgtctgt
      721 aatctgttgt tattgtttgt cactgtttat tcccatttgc tcctagttgc cgctggttta
      781 gaagctccat tcctgtactt gtacgctttg gtctatttcc tgcaatctat caactttgtt
      841 cgtatcatta tgagattgtg gttgtgctgg aaatgtcgtt ctaagaaccc gttgttatac
      901 gacgctaatt acttcctttg ttggcacacc aactgttacg attattgtat tccatataac
      961 tcggttactt cctctatagt tatcacctcc ggggacggta ccacaagtcc aatttcagaa
     1021 catgactacc aaattggtgg ttacactgaa aagtgggagt ctggtgtcaa agattgcgta
     1081 gtgttacaca gctattttac ttctgattac taccagttgt atagcaccca attgtccact
     1141 gacaccggag tggaacacgt cacatttttc atctacaata agatagtcga tgaacctgaa
     1201 gaacatgttc aaatacacac tattgacggc tctagcggtg tagttaaccc agttatggaa
     1261 ccaatctatg atgagccaac aacaacgacc tctgttccct tactcgtgcc gagaggttct
     1321 caccatcatc accatcatta aacaggcccc ttttcctttg tcgatatcat gtaattagtt
     1381 atgtcacgct tacattcacg ccctcctccc acatccgctc taaccgaaaa ggaaggagtt
     1441 agacaacctg aagtctaggt ccctatttat tttttttaat agttatgtta gtattaagaa
     1501 cgttatttat atttcaaatt tttctttttt ttctgtacaa agcggccgcc gaaccactag
     1561 tgcactgcag tacacggttt ccttgaaatt tttttgattc ggtaatctcc gaacagaagg
     1621 aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag
     1681 aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacg tgcaggaaac
     1741 gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg
     1801 ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg
     1861 atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt
     1921 tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc
     1981 taaaggcatt atccgccaag tacaattttt tactcttcga ggacagaaaa tttgctgaca
     2041 ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag
     2101 acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg
     2161 cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg
     2221 gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag
     2281 attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt
     2341 ggttgattat gacacccggt gtgggtttag atgacaaggg agatgcattg ggtcaacagt
     2401 atagaaccgt ggatgatgtg gtttctacag gatctgacat tattattgtt ggaagaggac
     2461 tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg
     2521 aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat
     2581 gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta cccgagtaac
     2641 gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa
     2701 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc gctattttac
     2761 caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga gcgctaattt
     2821 ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg agagcgctat
     2881 tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc cgagagcgct
     2941 atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc tataatgcag
     3001 tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct actttggtgt
     3061 ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg attactagcg
     3121 aagctgcggg tgcatttttt caagataaag gcatccccga ttatattcta taccgatgtg
     3181 gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat tggtcagaaa
     3241 attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat gtttacattt
     3301 tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt gtctaaagag
     3361 taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc
     3421 gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc aaagagatac
     3481 ttttgagcaa tgtttgtgga agcggtattc gcaatatttt agtagctcgt tacagtccgg
     3541 tgcgtttttg gttttttgaa agtgcgtcat cagagcgctt ttggttttca aaagcgctct
     3601 gaagttccta tactttctag ctagagaata ggaacttccc gagcggccgc gtgttacaag
     3661 cttgactcca gcgtaactgg actgcaatca actcactggc tcaccttcac gggtgggcct
     3721 ttcttcggta gaaaatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3781 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3841 caactctttt tccgaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3901 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3961 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4021 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4081 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4141 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4201 ggtcggaaca ggagagcgca cgagggagct tccaggggta aacgcctggt atctttatag
     4261 tcctgtcggg tttcgccacc tctgacttga gcatcgattt ttgtgatgct cgtcaggggg
     4321 gcggagccta tggaaaaacg ccagcaacgc agaaaggccc acccgaaggt gagccaggtg
     4381 attacatttg ggccctcatc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
     4441 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
     4501 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
     4561 gttaagggcg gttttttcct gtttggtcac ttaccaatgc ttaatcagtg aggcacctat
     4621 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4681 tacgatgcgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4741 ctcaccggct ccagatttat cagcaataaa ccagccagcc gggagggccg agcgcagaag
     4801 tgatcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4861 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4921 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4981 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgc
     5041 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5101 tactgtcatg ccatccgtga gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5161 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5221 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgtaa
     5281 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5341 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5401 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca ttttagcttc
     5461 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     5521 atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa aagcctccgg tcggaggctt
     5581 ttgactttct gctatggagg tcaggtatga tttaaatggt cagttaga
//
