LOCUS       Exported                4846 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp11.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046432
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4846)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4846)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4846
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             473..490
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             491..556
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(601..619)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             970..1773
                     /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      1856..2838
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2916..2945
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2967..3554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      3566..3593
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3617..3808)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3809..4669)
                     /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     4432..4451
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4670..4772)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      4773..4804
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atgtccgctg acgcacaatc attcctaaat gggtttgctg tactcgtgcc
      481 gagaggttct gattataaag atcatgatgg tgattataaa gatcatgata ttgattataa
      541 agatgatgat gataaataaa caggcccctt ttcctttgtc gatatcatgt aattagttat
      601 gtcacgctta cattcacgcc ctcctcccac atccgctcta accgaaaagg aaggagttag
      661 acaacctgaa gtctaggtcc ctatttattt tttttaatag ttatgttagt attaagaacg
      721 ttatttatat ttcaaatttt tctttttttt ctgtacaaag cggccgccga accactagtg
      781 cactgcagta cacggtttcc ttgaaatttt tttgattcgg taatctccga acagaaggaa
      841 gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa
      901 acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaacgtg caggaaacga
      961 agataaatca tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt
     1021 gctgccaagc tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat
     1081 gttcgtacca ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta
     1141 ctaaaaacac atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta
     1201 aaggcattat ccgccaagta caatttttta ctcttcgagg acagaaaatt tgctgacatt
     1261 ggtaatacag tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac
     1321 attacgaatg cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca
     1381 gaagaagtaa caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc
     1441 tccctatcta ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat
     1501 tttgttatcg gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg
     1561 ttgattatga cacccggtgt gggtttagat gacaagggag atgcattggg tcaacagtat
     1621 agaaccgtgg atgatgtggt ttctacagga tctgacatta ttattgttgg aagaggacta
     1681 tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa
     1741 gcatatttga gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt
     1801 atactaaact cacaaattag agcttcaatt taattatatc agttattacc cgagtaacga
     1861 agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac
     1921 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca
     1981 acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt
     2041 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt
     2101 taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat
     2161 ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc
     2221 tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct
     2281 attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa
     2341 gctgcgggtg cattttttca agataaaggc atccccgatt atattctata ccgatgtgga
     2401 ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat
     2461 tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc
     2521 gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta
     2581 atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga
     2641 aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt
     2701 ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg
     2761 cgtttttggt tttttgaaag tgcgtcatca gagcgctttt ggttttcaaa agcgctctga
     2821 agttcctata ctttctagct agagaatagg aacttcccga gcggccgcgt gttacaagct
     2881 tgactccagc gtaactggac tgcaatcaac tcactggctc accttcacgg gtgggccttt
     2941 cttcggtaga aaatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3001 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3061 actctttttc cgaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3121 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3181 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3241 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3301 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3361 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3421 tcggaacagg agagcgcacg agggagcttc caggggtaaa cgcctggtat ctttatagtc
     3481 ctgtcgggtt tcgccacctc tgacttgagc atcgattttt gtgatgctcg tcaggggggc
     3541 ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac ccgaaggtga gccaggtgat
     3601 tacatttggg ccctcatcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     3661 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     3721 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     3781 taagggcggt tttttcctgt ttggtcactt accaatgctt aatcagtgag gcacctatct
     3841 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     3901 cgatgcggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     3961 caccggctcc agatttatca gcaataaacc agccagccgg gagggccgag cgcagaagtg
     4021 atcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4081 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4141 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4201 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgcca
     4261 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4321 ctgtcatgcc atccgtgaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4381 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4441 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgtaaac
     4501 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4561 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4621 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcatt ttagcttcct
     4681 tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat
     4741 ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa gcctccggtc ggaggctttt
     4801 gactttctgc tatggaggtc aggtatgatt taaatggtca gttaga
//
