LOCUS       Exported                5349 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 leader protein.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046340
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5349)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5349)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5349
                     /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             1018..1035
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1036..1059
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1104..1122)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1473..2276
                     /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      2359..3341
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3419..3448
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3470..4057
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4069..4096
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4120..4311)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4312..5172)
                     /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     4935..4954
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5173..5275)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5276..5307
                     /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 gaatctttgg tccctggatt taacgagaaa acccacgtac agctatccct tcccgtgttg
      541 caagttcgtg acgttctggt tcggggtttc ggtgattcag tcgaagaagt cttgagtgaa
      601 gccaggcaac atttaaagga tggtacttgt ggcttagttg aagtcgaaaa gggtgtgttg
      661 ccacaacttg agcaaccata cgttttcatt aagagatctg acgcaagaac agcgccacac
      721 ggtcatgtta tggttgaatt ggtcgctgaa ttggagggta tacaatatgg cagatcgggt
      781 gaaactttag gtgtattggt cccacacgta ggtgaaatcc ctgttgctta cagaaaagtt
      841 ttattaagaa agaatggtaa caagggtgct ggtggacact cctacggagc cgatctgaag
      901 tcttttgact tgggtgatga attgggtact gatccatatg aagacttcca agaaaattgg
      961 aacaccaaac atagctctgg tgttacgcgt gaattgatga gagaacttaa tggtggtctc
     1021 gtgccgagag gttcttggag ccatccgcag tttgaaaaat aaacaggccc cttttccttt
     1081 gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc cacatccgct
     1141 ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta ttttttttaa
     1201 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca
     1261 aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat ttttttgatt
     1321 cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga ttggtatata
     1381 tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc caactgcaca
     1441 gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat aaggaacgtg
     1501 ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac gaaaagcaaa
     1561 caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag ttagttgaag
     1621 cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact gatttttcca
     1681 tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt ttactcttcg
     1741 aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct gcgggtgtat
     1801 acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc ccaggtattg
     1861 ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc cttttgatgt
     1921 tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag ggtactgttg
     1981 acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga gacatgggtg
     2041 gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta gatgacaagg
     2101 gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca ggatctgaca
     2161 ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac
     2221 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa
     2281 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
     2341 atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac
     2401 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     2461 aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat
     2521 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     2581 aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca
     2641 aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc
     2701 tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt
     2761 agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc
     2821 cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg
     2881 attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga
     2941 tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac
     3001 gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact
     3061 acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt
     3121 ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag
     3181 agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt
     3241 tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca tcagagcgct
     3301 tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat aggaacttcc
     3361 cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc aactcactgg
     3421 ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct tgagatcctt
     3481 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3541 gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca gcagagcgca
     3601 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3661 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3721 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3781 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3841 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3901 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggt
     3961 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt
     4021 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cagaaaggcc
     4081 cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt caccgtcatc
     4141 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     4201 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     4261 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca cttaccaatg
     4321 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4381 actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc ccagtgctgc
     4441 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4501 cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc agtctattaa
     4561 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4621 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4681 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4741 cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac tcatggttat
     4801 ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt ctgtgactgg
     4861 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4921 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4981 aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5041 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5101 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5161 ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
     5221 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaagtca
     5281 aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg atttaaatgg
     5341 tcagttaga
//
