LOCUS       Exported                5217 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp10.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046463
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5217)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5217)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5217
                     /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"
     CDS             910..927
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(972..990)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1341..2144
                     /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      2227..3209
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3287..3316
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3338..3925
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      3937..3964
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3988..4179)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4180..5040)
                     /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     4803..4822
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5041..5143)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5144..5175
                     /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 agaggttctc accatcatca ccatcattaa acaggcccct tttcctttgt cgatatcatg
      961 taattagtta tgtcacgctt acattcacgc cctcctccca catccgctct aaccgaaaag
     1021 gaaggagtta gacaacctga agtctaggtc cctatttatt ttttttaata gttatgttag
     1081 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaaa gcggccgccg
     1141 aaccactagt gcactgcagt acacggtttc cttgaaattt ttttgattcg gtaatctccg
     1201 aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata cgcatatgta
     1261 gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga acaaaaacgt
     1321 gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct gctactcatc
     1381 ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca aacttgtgtg
     1441 cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca ttaggtccca
     1501 aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg gagggcacag
     1561 ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgag gacagaaaat
     1621 ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac agaatagcag
     1681 aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt agcggtttga
     1741 agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta gcagaattgt
     1801 catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac attgcgaaga
     1861 gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga agagatgaag
     1921 gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga gatgcattgg
     1981 gtcaacagta tagaaccgtg gatgatgtgg tttctacagg atctgacatt attattgttg
     2041 gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag
     2101 caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact gtattataag
     2161 taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat cagttattac
     2221 ccgagtaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta
     2281 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg
     2341 ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag
     2401 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     2461 gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc
     2521 gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct
     2581 ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta
     2641 ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga
     2701 ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat tatattctat
     2761 accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt
     2821 ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg
     2881 tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg
     2941 tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt
     3001 tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca
     3061 aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt
     3121 acagtccggt gcgtttttgg ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa
     3181 aagcgctctg aagttcctat actttctagc tagagaatag gaacttcccg agcggccgcg
     3241 tgttacaagc ttgactccag cgtaactgga ctgcaatcaa ctcactggct caccttcacg
     3301 ggtgggcctt tcttcggtag aaaatcaaag gatcttcttg agatcctttt tttctgcgcg
     3361 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3421 aagagctacc aactcttttt ccgaggtaac tggcttcagc agagcgcaga taccaaatac
     3481 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3541 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3601 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3661 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3721 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3781 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccaggggtaa acgcctggta
     3841 tctttatagt cctgtcgggt ttcgccacct ctgacttgag catcgatttt tgtgatgctc
     3901 gtcagggggg cggagcctat ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg
     3961 agccaggtga ttacatttgg gccctcatca gaggttttca ccgtcatcac cgaaacgcgc
     4021 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     4081 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     4141 gcgggccatg ttaagggcgg ttttttcctg tttggtcact taccaatgct taatcagtga
     4201 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4261 gtagataact acgatgcggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4321 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg ggagggccga
     4381 gcgcagaagt gatcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4441 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4501 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4561 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4621 gatcgttgcc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4681 taattctctt actgtcatgc catccgtgag atgcttttct gtgactggtg agtactcaac
     4741 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4801 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4861 ggggcgtaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4921 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4981 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5041 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     5101 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa agcctccggt
     5161 cggaggcttt tgactttctg ctatggaggt caggtatgat ttaaatggtc agttaga
//
