LOCUS       Exported                4798 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046544
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4798)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4798)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4798
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        19..426
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             476..493
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             494..517
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(562..580)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             931..1734
                     /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      1817..2799
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2877..2906
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2928..3515
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      3527..3554
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3578..3769)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3770..4630)
                     /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     4393..4412
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4631..4733)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      4734..4765
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 tcagttagag cggccgcccc acacaccata gcttcaaaat gtttctactc cttttttact
       61 cttccagatt ttctcggact ccgcgcatcg ccgtaccact tcaaaacacc caagcacagc
      121 atactaaatt tcccctcttt cttcctctag ggtgtcgtta attacccgta ctaaaggttt
      181 ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt cgaaaaaggc aataaaaatt
      241 tttatcacgt ttctttttct tgaaaatttt tttttttgat ttttttctct ttcgatgacc
      301 tcccattgat atttaagtta ataaacggtc ttcaatttct caagtttcag tttcattttt
      361 cttgttctat tacaactttt tttacttctt gctcattaga aagaaagcat agcaatctaa
      421 tctaagtttt aattacaaaa tggctgatgc acaatcgttt ttaaacgggt ttgcgctcgt
      481 gccgagaggt tcttggagcc atccgcagtt tgaaaaataa acaggcccct tttcctttgt
      541 cgatatcatg taattagtta tgtcacgctt acattcacgc cctcctccca catccgctct
      601 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt ttttttaata
      661 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaaa
      721 gcggccgccg aaccactagt gcactgcagt acacggtttc cttgaaattt ttttgattcg
      781 gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata
      841 cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga
      901 acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct
      961 gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca
     1021 aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca
     1081 ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg
     1141 gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgag
     1201 gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac
     1261 agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt
     1321 agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta
     1381 gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac
     1441 attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga
     1501 agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga
     1561 gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg tttctacagg atctgacatt
     1621 attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt
     1681 tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact
     1741 gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat
     1801 cagttattac ccgagtaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc
     1861 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     1921 cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc
     1981 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     2041 tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa
     2101 aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt
     2161 tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag
     2221 aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg
     2281 cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat
     2341 tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg
     2401 attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt
     2461 ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac
     2521 aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt
     2581 agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag
     2641 atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta
     2701 gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcatc agagcgcttt
     2761 tggttttcaa aagcgctctg aagttcctat actttctagc tagagaatag gaacttcccg
     2821 agcggccgcg tgttacaagc ttgactccag cgtaactgga ctgcaatcaa ctcactggct
     2881 caccttcacg ggtgggcctt tcttcggtag aaaatcaaag gatcttcttg agatcctttt
     2941 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3001 ttgccggatc aagagctacc aactcttttt ccgaggtaac tggcttcagc agagcgcaga
     3061 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3121 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3181 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3241 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3301 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3361 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccaggggtaa
     3421 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag catcgatttt
     3481 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgca gaaaggccca
     3541 cccgaaggtg agccaggtga ttacatttgg gccctcatca gaggttttca ccgtcatcac
     3601 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     3661 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     3721 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact taccaatgct
     3781 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3841 tccccgtcgt gtagataact acgatgcggg agggcttacc atctggcccc agtgctgcaa
     3901 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3961 ggagggccga gcgcagaagt gatcctgcaa ctttatccgc ctccatccag tctattaatt
     4021 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4081 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4141 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4201 tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4261 cagcactgca taattctctt actgtcatgc catccgtgag atgcttttct gtgactggtg
     4321 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4381 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4441 aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4501 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4561 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4621 gaatactcat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     4681 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa
     4741 agcctccggt cggaggcttt tgactttctg ctatggaggt caggtatgat ttaaatgg
//
