LOCUS       Exported                4803 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046701
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4803)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4803)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4803
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        24..465
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    24..141
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     436..459
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(557..575)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             926..1729
                     /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      1812..2794
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2872..2901
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2923..3510
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      3522..3549
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3573..3764)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3765..4625)
                     /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     4388..4407
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4626..4728)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      4729..4760
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 cggccgcttg ctatattgaa gtacggatta gaagccgccg agcgggcgac agccctccga
       61 cggaagactc tcctccgtgc gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt
      121 gcctcgcgcc gcactgctcc gaacaataaa gattctacaa tactagcttt tatggttatg
      181 aagaggaaaa attggcagta acctggcccc acaaaccttc aaattaacga atcaaattaa
      241 caaccatagg atgataatgc gattagtttt ttagccttat ttctggggta attaatcagc
      301 gaagcgatga tttttgatct attaacagat atataaatgg aaaagctgca taaccacttt
      361 aactaatact ttcaacattt tcagtttgta ttacttctta ttcaaatgtc ataaaagtat
      421 caacaaaaaa ttgttaatat acctctatac tttaacgtca aggagaaaaa actataatgg
      481 ctgatgcaca atcgttttta aacgggtttg cgtaaacagg ccccttttcc tttgtcgata
      541 tcatgtaatt agttatgtca cgcttacatt cacgccctcc tcccacatcc gctctaaccg
      601 aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt taatagttat
      661 gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt acaaagcggc
      721 cgccgaacca ctagtgcact gcagtacacg gtttccttga aatttttttg attcggtaat
      781 ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat
      841 atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa
      901 aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac
      961 tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt
     1021 gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg
     1081 tcccaaaatt tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg
     1141 cacagttaag ccgctaaagg cattatccgc caagtacaat tttttactct tcgaggacag
     1201 aaaatttgct gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat
     1261 agcagaatgg gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg
     1321 tttgaagcag gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga
     1381 attgtcatgc aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc
     1441 gaagagcgac aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga
     1501 tgaaggttac gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagatgc
     1561 attgggtcaa cagtatagaa ccgtggatga tgtggtttct acaggatctg acattattat
     1621 tgttggaaga ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag
     1681 aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt
     1741 ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt
     1801 attacccgag taacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag
     1861 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     1921 aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc
     1981 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     2041 cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc
     2101 atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc
     2161 gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa
     2221 ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt
     2281 actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat
     2341 tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct
     2401 tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg
     2461 aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt
     2521 ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg
     2581 caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata
     2641 tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc
     2701 tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc gcttttggtt
     2761 ttcaaaagcg ctctgaagtt cctatacttt ctagctagag aataggaact tcccgagcgg
     2821 ccgcgtgtta caagcttgac tccagcgtaa ctggactgca atcaactcac tggctcacct
     2881 tcacgggtgg gcctttcttc ggtagaaaat caaaggatct tcttgagatc ctttttttct
     2941 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3001 ggatcaagag ctaccaactc tttttccgag gtaactggct tcagcagagc gcagatacca
     3061 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3121 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3181 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3241 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3301 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3361 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg ggtaaacgcc
     3421 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcatcg atttttgtga
     3481 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcagaaag gcccacccga
     3541 aggtgagcca ggtgattaca tttgggccct catcagaggt tttcaccgtc atcaccgaaa
     3601 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     3661 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     3721 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcacttacca atgcttaatc
     3781 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3841 gtcgtgtaga taactacgat gcgggagggc ttaccatctg gccccagtgc tgcaatgata
     3901 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccgggagg
     3961 gccgagcgca gaagtgatcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4021 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4081 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4141 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4201 cctccgatcg ttgccagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4261 ctgcataatt ctcttactgt catgccatcc gtgagatgct tttctgtgac tggtgagtac
     4321 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4381 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4441 tcttcggggc gtaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4501 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4561 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4621 ctcattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt
     4681 gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaag tcaaaagcct
     4741 ccggtcggag gcttttgact ttctgctatg gaggtcaggt atgatttaaa tggtcagtta
     4801 gag
//
