LOCUS       Exported                6297 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp10.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046398
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6297)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6297)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6297
                     /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             892..909
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2052..2070)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2421..3224
                     /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      3307..4289
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4367..4396
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4418..5005
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      5017..5044
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5068..5259)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5260..6120)
                     /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     5883..5902
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6121..6223)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      6224..6255
                     /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 ggtaatgcaa cagaagtgcc tgccaattca actgtattat ctttctgtgc ttttgctgta
      541 gatgctgcta aagcttacaa agattatcta gctagtgggg gacaaccaat cactaattgt
      601 gttaagatgt tgtgtacaca cactggtact ggtcaggcaa taacagttac accggaagcc
      661 aatatggatc aagaatcctt tggtggtgca tcgtgttgtc tgtactgccg ttgccacata
      721 gatcatccaa atcctaaagg attttgtgac ttaaaaggta agtatgtaca aatacctaca
      781 acttgtgcta atgaccctgt gggttttaca cttaaaaaca cagtctgtac cgtctgcggt
      841 atgtggaaag gttatggctg tagttgtgat caactccgcg aacccatgct tctcgtgccg
      901 agaggttcta aaatcgagga aggtaagcta gtcatttgga taaatggaga taagggttac
      961 aacggtttgg cagaagtagg caagaaattt gaaaaggaca caggtatcaa ggtgacggtt
     1021 gaacatccag acaagttaga agaaaaattc cctcaagttg ctgctactgg tgatggtccc
     1081 gacattattt tctgggccca cgatcgtttt ggtggatatg ctcagtcagg tttgctggct
     1141 gagatcaccc cagacaaggc cttccaagat aaattgtacc cattcacttg ggatgctgtc
     1201 agatacaacg gtaagcttat agcatatcca attgctgttg aagccttatc gttgatctac
     1261 aataaggact tgttacctaa cccaccaaag acatgggaag aaattccagc tttggataaa
     1321 gagctaaagg ccaagggaaa aagcgctttg atgtttaact tacaagaacc ttatttcacc
     1381 tggccattaa tcgctgctga cggtggttac gcatttaaat acgaaaacgg taagtatgac
     1441 attaaggatg tcggcgttga taatgctggt gcaaaggctg gtttgacctt cttggttgat
     1501 ttaataaaga ataaacacat gaacgctgac actgactact ctatcgccga agccgctttc
     1561 aacaagggtg aaactgctat gaccataaat ggtccatggg cctggagtaa cattgatact
     1621 tctaaagtaa actatggtgt cactgttttg cctacattca aaggccaacc atccaagccc
     1681 tttgtcggtg ttctatccgc tggcatcaat gctgcttctc caaacaagga gctagctaaa
     1741 gaatttttgg aaaattactt gttaactgac gaaggtttgg aggccgttaa caaggacaag
     1801 ccattgggtg ctgtcgcttt gaaatcttac gaagaagaat tggctaagga tcccagaatt
     1861 gccgctacca tggagaacgc ccaaaagggt gaaattatgc caaacatccc acaaatgtca
     1921 gccttctggt acgcagttag gactgccgtg attaatgctg cttccggtag acaaaccgtc
     1981 gacgaagcat taaaagatgc tcaaacgtaa acaggcccct tttcctttgt cgatatcatg
     2041 taattagtta tgtcacgctt acattcacgc cctcctccca catccgctct aaccgaaaag
     2101 gaaggagtta gacaacctga agtctaggtc cctatttatt ttttttaata gttatgttag
     2161 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaaa gcggccgccg
     2221 aaccactagt gcactgcagt acacggtttc cttgaaattt ttttgattcg gtaatctccg
     2281 aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata cgcatatgta
     2341 gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga acaaaaacgt
     2401 gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct gctactcatc
     2461 ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca aacttgtgtg
     2521 cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca ttaggtccca
     2581 aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg gagggcacag
     2641 ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgag gacagaaaat
     2701 ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac agaatagcag
     2761 aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt agcggtttga
     2821 agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta gcagaattgt
     2881 catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac attgcgaaga
     2941 gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga agagatgaag
     3001 gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga gatgcattgg
     3061 gtcaacagta tagaaccgtg gatgatgtgg tttctacagg atctgacatt attattgttg
     3121 gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag
     3181 caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact gtattataag
     3241 taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat cagttattac
     3301 ccgagtaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta
     3361 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg
     3421 ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag
     3481 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     3541 gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc
     3601 gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct
     3661 ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta
     3721 ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga
     3781 ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat tatattctat
     3841 accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt
     3901 ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg
     3961 tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg
     4021 tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt
     4081 tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca
     4141 aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt
     4201 acagtccggt gcgtttttgg ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa
     4261 aagcgctctg aagttcctat actttctagc tagagaatag gaacttcccg agcggccgcg
     4321 tgttacaagc ttgactccag cgtaactgga ctgcaatcaa ctcactggct caccttcacg
     4381 ggtgggcctt tcttcggtag aaaatcaaag gatcttcttg agatcctttt tttctgcgcg
     4441 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4501 aagagctacc aactcttttt ccgaggtaac tggcttcagc agagcgcaga taccaaatac
     4561 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4621 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4681 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4741 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4801 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4861 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccaggggtaa acgcctggta
     4921 tctttatagt cctgtcgggt ttcgccacct ctgacttgag catcgatttt tgtgatgctc
     4981 gtcagggggg cggagcctat ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg
     5041 agccaggtga ttacatttgg gccctcatca gaggttttca ccgtcatcac cgaaacgcgc
     5101 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     5161 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     5221 gcgggccatg ttaagggcgg ttttttcctg tttggtcact taccaatgct taatcagtga
     5281 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5341 gtagataact acgatgcggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5401 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg ggagggccga
     5461 gcgcagaagt gatcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5521 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5581 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5641 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5701 gatcgttgcc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5761 taattctctt actgtcatgc catccgtgag atgcttttct gtgactggtg agtactcaac
     5821 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5881 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5941 ggggcgtaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6001 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6061 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6121 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     6181 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa agcctccggt
     6241 cggaggcttt tgactttctg ctatggaggt caggtatgat ttaaatggtc agttaga
//
