LOCUS       Exported                7555 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 RNA-dependent RNA 
            polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046258
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7555)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7555)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7555
                     /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             3224..3241
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             3242..3265
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3310..3328)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3679..4482
                     /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      4565..5547
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5625..5654
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5676..6263
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      6275..6302
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6326..6517)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6518..7378)
                     /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     7141..7160
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7379..7481)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      7482..7513
                     /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 atggctgatg cacaatcgtt tttaaaccgg gtttgcggtg taagtgcagc
      481 ccgtcttaca ccgtgcggca caggcactag tactgatgtc gtatacaggg cttttgacat
      541 ctacaatgat aaagtagctg gttttgctaa attcctaaaa actaattgtt gtcgcttcca
      601 agaaaaggac gaagatgaca atttaattga ttcttacttt gtagttaaga gacacacttt
      661 ctctaactac caacatgaag aaacaattta taatttactt aaggattgtc cagctgttgc
      721 taaacatgac ttctttaagt ttagaataga cggtgacatg gtaccacata tatcacgtca
      781 acgtcttact aaatacacaa tggcagacct cgtctatgct ttaaggcatt ttgatgaagg
      841 taattgtgac acattaaaag aaatacttgt cacatacaat tgttgtgatg atgattattt
      901 caataaaaag gactggtatg attttgtaga aaacccagat atattacgcg tatacgccaa
      961 cttaggtgaa cgtgtacgcc aagctttgtt aaaaacagta caattctgtg atgccatgcg
     1021 aaatgctggt attgttggtg tactgacatt agataatcaa gatctcaatg gtaactggta
     1081 tgatttcggt gatttcatac aaaccacgcc aggtagtgga gttcctgttg tagattctta
     1141 ttattcattg ttaatgccta tattaacctt gaccagggct ttaactgcag agtcacatgt
     1201 tgacactgac ttaacaaagc cttacattaa gtgggatttg ttaaaatatg acttcacgga
     1261 agagaggtta aaactctttg accgttattt taaatattgg gatcagacat accacccaaa
     1321 ttgtgttaac tgtttggatg acagatgcat tctgcattgt gcaaacttta atgttttatt
     1381 ctctacagtg ttcccaccta caagttttgg accactagtg agaaaaatat ttgttgatgg
     1441 tgttccattt gtagtttcaa ctggatacca cttcagagag ctaggtgttg tacataatca
     1501 ggatgtaaac ttacatagct ctagacttag ttttaaggaa ttacttgtgt atgctgctga
     1561 ccctgctatg cacgctgctt ctggtaatct attactagat aaacgcacta cgtgcttttc
     1621 agtagctgca cttactaaca atgttgcttt tcaaactgtc aaacccggta attttaacaa
     1681 agacttctat gactttgctg tgtctaaggg tttctttaag gaaggaagtt ctgttgaatt
     1741 aaaacacttc ttctttgctc aggatggtaa tgctgctatc agcgattatg actactatcg
     1801 ttataatcta ccaacaatgt gtgatatcag acaactacta tttgtagttg aagttgttga
     1861 taagtacttt gattgttacg atggtggctg tattaatgct aaccaagtca tcgtcaacaa
     1921 cctagacaaa tcagctggtt ttccatttaa taaatggggt aaggctagac tttattatga
     1981 ttcaatgagt tatgaggatc aagatgcact tttcgcatat acaaaacgta atgtcatccc
     2041 tactataact caaatgaatc ttaagtatgc cattagtgca aagaatagag ctcgcaccgt
     2101 agctggtgtc tctatctgta gtactatgac caatagacag tttcatcaaa aattattgaa
     2161 atcaatagcc gccactagag gagctactgt agtaattgga acaagcaaat tctatggtgg
     2221 ttggcacaac atgttaaaaa ctgtttatag tgatgtagaa aaccctcacc ttatgggttg
     2281 ggattatcct aaatgtgata gagccatgcc taacatgctt agaattatgg cctcacttgt
     2341 tcttgctcgc aaacatacaa cgtgttgtag cttgtcacac cgtttctata gattagctaa
     2401 tgagtgtgct caagtattga gtgaaatggt catgtgtggc ggttcactat atgttaaacc
     2461 aggtggaacc tcatcaggag atgccacaac tgcttatgct aatagtgttt ttaacatttg
     2521 tcaagctgtc acggccaatg ttaatgcact tttatctact gatggtaaca aaattgccga
     2581 taagtatgtc cgcaatttac aacacagact ttatgagtgt ctctatagaa atagagatgt
     2641 tgacacagac tttgtgaatg agttttacgc atatttgcgt aaacatttct caatgatgat
     2701 actctctgac gatgctgttg tgtgtttcaa tagcacttat gcatctcaag gtctagtggc
     2761 tagcataaag aactttaagt cagttcttta ttatcaaaac aatgttttta tgtctgaagc
     2821 aaaatgttgg actgagactg accttactaa aggacctcat gaattttgct ctcaacatac
     2881 aatgctagtt aaacagggtg atgattatgt gtaccttcct tacccagatc catcaagaat
     2941 cctaggggcc ggctgttttg tagatgatat cgtaaaaaca gatggtacac ttatgattga
     3001 acggttcgtg tctttagcta tagatgctta cccacttact aaacatccta atcaggagta
     3061 tgctgatgtc tttcatttgt acttacaata cataagaaag ctacatgatg agttaacagg
     3121 acacatgtta gacatgtatt ctgttatgct tactaatgat aacacttcaa ggtattggga
     3181 acctgagttt tatgaggcta tgtacacacc gcatacagtc ttactcgtgc cgagaggttc
     3241 ttggagccat ccgcagtttg aaaaataaac aggccccttt tcctttgtcg atatcatgta
     3301 attagttatg tcacgcttac attcacgccc tcctcccaca tccgctctaa ccgaaaagga
     3361 aggagttaga caacctgaag tctaggtccc tatttatttt ttttaatagt tatgttagta
     3421 ttaagaacgt tatttatatt tcaaattttt cttttttttc tgtacaaagc ggccgccgaa
     3481 ccactagtgc actgcagtac acggtttcct tgaaattttt ttgattcggt aatctccgaa
     3541 cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt
     3601 gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacgtgc
     3661 aggaaacgaa gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct
     3721 agtcctgttg ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct
     3781 tcattggatg ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa
     3841 atttgtttac taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt
     3901 aagccgctaa aggcattatc cgccaagtac aattttttac tcttcgagga cagaaaattt
     3961 gctgacattg gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa
     4021 tgggcagaca ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag
     4081 caggcggcag aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca
     4141 tgcaagggct ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc
     4201 gacaaagatt ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt
     4261 tacgattggt tgattatgac acccggtgtg ggtttagatg acaagggaga tgcattgggt
     4321 caacagtata gaaccgtgga tgatgtggtt tctacaggat ctgacattat tattgttgga
     4381 agaggactat ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca
     4441 ggctgggaag catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta
     4501 aatgcatgta tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc
     4561 gagtaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat
     4621 ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct
     4681 attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg
     4741 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga
     4801 gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga
     4861 gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat
     4921 aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact
     4981 ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt
     5041 actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac
     5101 cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg
     5161 tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt
     5221 tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc
     5281 taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc
     5341 aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa
     5401 gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac
     5461 agtccggtgc gtttttggtt ttttgaaagt gcgtcatcag agcgcttttg gttttcaaaa
     5521 gcgctctgaa gttcctatac tttctagcta gagaatagga acttcccgag cggccgcgtg
     5581 ttacaagctt gactccagcg taactggact gcaatcaact cactggctca ccttcacggg
     5641 tgggcctttc ttcggtagaa aatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5701 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5761 gagctaccaa ctctttttcc gaggtaactg gcttcagcag agcgcagata ccaaatactg
     5821 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5881 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5941 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6001 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6061 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6121 gcggcagggt cggaacagga gagcgcacga gggagcttcc aggggtaaac gcctggtatc
     6181 tttatagtcc tgtcgggttt cgccacctct gacttgagca tcgatttttg tgatgctcgt
     6241 caggggggcg gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc cgaaggtgag
     6301 ccaggtgatt acatttgggc cctcatcaga ggttttcacc gtcatcaccg aaacgcgcga
     6361 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     6421 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     6481 gggccatgtt aagggcggtt ttttcctgtt tggtcactta ccaatgctta atcagtgagg
     6541 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6601 agataactac gatgcgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6661 acccacgctc accggctcca gatttatcag caataaacca gccagccggg agggccgagc
     6721 gcagaagtga tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6781 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6841 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6901 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6961 tcgttgccag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7021 attctcttac tgtcatgcca tccgtgagat gcttttctgt gactggtgag tactcaacca
     7081 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7141 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7201 ggcgtaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7261 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7321 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcattt
     7381 tagcttcctt agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta
     7441 tttcattatg gtgaaagttg gaacctctta cgtgccgatc aagtcaaaag cctccggtcg
     7501 gaggcttttg actttctgct atggaggtca ggtatgattt aaatggtcag ttaga
//
