LOCUS       Exported                6733 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast expression plasmid for SARS-CoV-2 2'-O-ribose 
            methyltransferase.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046279
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6733)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6733)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6733
                     /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             1328..1345
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2488..2506)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2857..3660
                     /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      3743..4725
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4803..4832
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4854..5441
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      5453..5480
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5504..5695)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5696..6556)
                     /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     6319..6338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6557..6659)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      6660..6691
                     /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 atgtcctctc aagcctggca gcctggtgtg gctatgccca atttgtataa
      481 gatgcaaaga atgcttctgg agaaatgcga tttgcaaaac tacggagaca gcgctacatt
      541 accaaagggt attatgatga acgtagcaaa gtacacccaa ttgtgtcaat acttaaatac
      601 gttgactttg gctgttccgt ataacatgcg tgtcatacac tttggggccg gttcagacaa
      661 aggcgtcgct ccaggtaccg cagttctaag gcaatggctc ccaactggta ctctattggt
      721 tgattctgat ctgaacgact tcgtcagtga tgctgactcc actttaatcg gtgattgtgc
      781 taccgttcat acggcgaaca agtgggactt gattatttcg gatatgtacg acccaaagac
      841 caaaaatgtt acaaaggaaa atgattctaa agaaggtttc tttacttata tctgtggttt
      901 catccagcaa aagcttgcct tgggcggttc tgtggctatt aagatcactg aacactcctg
      961 gaacgccgac ttatacaaat tgatgggtca tttcgcttgg tggaccgcgt ttgttaccaa
     1021 cgttaacgct agttcttcgg aagcctttct catcggttgt aattatttag gaaagccacg
     1081 cgaacaaatt gatggttacg tcatgcacgc taactacatt ttctggagaa atactaaccc
     1141 tatacaattg tcaagctata gtttattcga catgtcaaag tttccattga agttgagagg
     1201 tacagctgtc atgtccttga aggaagggca gatcaacgat atgatcttat ctttgctgtc
     1261 taaaggtaga ctcattatta gagaaaataa tcgggttgtt atctcttctg atgtgctagt
     1321 caacaacctc gtgccgagag gttctaaaat cgaggaaggt aagctagtca tttggataaa
     1381 tggagataag ggttacaacg gtttggcaga agtaggcaag aaatttgaaa aggacacagg
     1441 tatcaaggtg acggttgaac atccagacaa gttagaagaa aaattccctc aagttgctgc
     1501 tactggtgat ggtcccgaca ttattttctg ggcccacgat cgttttggtg gatatgctca
     1561 gtcaggtttg ctggctgaga tcaccccaga caaggccttc caagataaat tgtacccatt
     1621 cacttgggat gctgtcagat acaacggtaa gcttatagca tatccaattg ctgttgaagc
     1681 cttatcgttg atctacaata aggacttgtt acctaaccca ccaaagacat gggaagaaat
     1741 tccagctttg gataaagagc taaaggccaa gggaaaaagc gctttgatgt ttaacttaca
     1801 agaaccttat ttcacctggc cattaatcgc tgctgacggt ggttacgcat ttaaatacga
     1861 aaacggtaag tatgacatta aggatgtcgg cgttgataat gctggtgcaa aggctggttt
     1921 gaccttcttg gttgatttaa taaagaataa acacatgaac gctgacactg actactctat
     1981 cgccgaagcc gctttcaaca agggtgaaac tgctatgacc ataaatggtc catgggcctg
     2041 gagtaacatt gatacttcta aagtaaacta tggtgtcact gttttgccta cattcaaagg
     2101 ccaaccatcc aagccctttg tcggtgttct atccgctggc atcaatgctg cttctccaaa
     2161 caaggagcta gctaaagaat ttttggaaaa ttacttgtta actgacgaag gtttggaggc
     2221 cgttaacaag gacaagccat tgggtgctgt cgctttgaaa tcttacgaag aagaattggc
     2281 taaggatccc agaattgccg ctaccatgga gaacgcccaa aagggtgaaa ttatgccaaa
     2341 catcccacaa atgtcagcct tctggtacgc agttaggact gccgtgatta atgctgcttc
     2401 cggtagacaa accgtcgacg aagcattaaa agatgctcaa acgtaaacag gccccttttc
     2461 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc
     2521 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     2581 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     2641 tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt
     2701 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
     2761 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
     2821 cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa
     2881 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag
     2941 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt
     3001 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt
     3061 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc
     3121 ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt
     3181 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt
     3241 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg
     3301 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact
     3361 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg
     3421 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac
     3481 aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct
     3541 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt
     3601 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
     3661 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
     3721 ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg
     3781 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     3841 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     3901 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     3961 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     4021 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     4081 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     4141 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     4201 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     4261 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     4321 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     4381 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     4441 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     4501 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     4561 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     4621 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag
     4681 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
     4741 ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca
     4801 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat
     4861 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4921 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag
     4981 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5041 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5101 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5161 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5221 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     5281 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5341 gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc
     5401 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa
     5461 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt
     5521 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     5581 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     5641 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc
     5701 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5761 cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg
     5821 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5881 cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta
     5941 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6001 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6061 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6121 gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg
     6181 ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga
     6241 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     6301 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     6361 ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt
     6421 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6481 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6541 aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     6601 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     6661 gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa
     6721 atggtcagtt aga
//
