LOCUS       Exported                7642 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 helicase.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046209
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7642)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7642)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7642
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             2237..2254
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(3397..3415)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3766..4569
                     /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      4652..5634
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5712..5741
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5763..6350
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      6362..6389
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6413..6604)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6605..7465)
                     /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     7228..7247
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7466..7568)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      7569..7600
                     /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 ttttnnnnnn
      241 nnnctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atggcagtcg gcgcttgtgt attatgcaac tcccagacgt cgttgagatg
      481 tggtgcttgt attcgtagac ctttcctttg ctgtaaatgt tgctatgacc acgttatctc
      541 aacttctcat aagctagttc tgagcgtcaa tccatacgtg tgtaacgccc ccgggtgtga
      601 tgttacagac gttacccaat tgtacttggg aggtatgtct tactattgta agtcccacaa
      661 gccacctata tcttttccat tatgcgctaa cggtcaagtc ttcggtttgt acaaaaatac
      721 ctgtgttggt agtgataatg ttaccgactt taacgccatt gctacttgtg attggactaa
      781 cgcgggtgat tatatcttgg ctaacacttg taccgagagg cttaagttgt tcgccgctga
      841 aaccttaaag gctacggaag aaacattcaa attgagttac ggtatcgcta ctgtcagaga
      901 agttctatct gatagagaat tgcatctctc ctgggaggtg ggtaagccaa gaccaccgtt
      961 gaaccggaat tacgtcttta ctggatacag ggttactaag aactctaagg ttcaaattgg
     1021 ggaatatacc tttgaaaagg gcgactacgg tgacgccgta gtctatagag gtaccaccac
     1081 ttacaaattg aacgtcggtg actatttcgt tttaaccagc cacacagtga tgccactgtc
     1141 agcgccaaca ttagttccgc aagagcatta cgttagaatt actggtttgt atccaactct
     1201 aaatatctct gatgaatttt cctccaatgt ggctaactac caaaaagtag gtatgcaaaa
     1261 gtactctacc ttgcaaggtc cacctggtac tgggaagtcg cacttcgcta taggtttggc
     1321 actgtactac ccatctgcta gaattgttta cacggcatgt tctcatgctg ctgtagacgc
     1381 cttgtgtgaa aaggccttga aatatttacc cattgataaa tgttcaagaa ttatcccagc
     1441 acgagctcgt gttgaatgtt tcgacaagtt taaggtcaac agcacactag aacaatacgt
     1501 cttctgcact gttaatgctt tgccagaaac gactgctgat atcgtcgtct ttgatgaaat
     1561 ctcgatggcc actaactatg acttatccgt agttaatgct agacttcgtg caaagcacta
     1621 cgtctatatt ggtgatccag ctcagctgcc tgcaccaaga actttgttaa ccaagggtac
     1681 cttggaacca gaatacttca actctgtgtg tcgcctgatg aagacaatcg gccctgatat
     1741 gttcctaggt acatgtagac gttgtccagc tgaaattgtg gacacggtca gtgccttggt
     1801 ttacgataac aaattaaagg cgcataagga caaaagcgct caatgtttca agatgtttta
     1861 caagggtgtc attactcacg atgttagttc ggccatcaat agaccccaaa taggtgttgt
     1921 tagagaattt ttgacaagga acccagcatg gagaaaggct gttttcatat ccccatacaa
     1981 ttctcaaaac gcggtagcct caaaaatctt gggtcttccc actcagaccg ttgattcttc
     2041 ccaaggaagt gaatatgact acgtgatttt cactcaaacc accgagactg cccactcttg
     2101 caacgtcaat cgtttcaacg ttgctattac gagagccaag gttggcatct tgtgtatcat
     2161 gtctgataga gacttgtacg ataaattaca gtttacatcc ttagaaatac caagacgaaa
     2221 cgtcgctact ttgcaactcg tgccgagagg ttctaaaatc gaggaaggta agctagtcat
     2281 ttggataaat ggagataagg gttacaacgg tttggcagaa gtaggcaaga aatttgaaaa
     2341 ggacacaggt atcaaggtga cggttgaaca tccagacaag ttagaagaaa aattccctca
     2401 agttgctgct actggtgatg gtcccgacat tattttctgg gcccacgatc gttttggtgg
     2461 atatgctcag tcaggtttgc tggctgagat caccccagac aaggccttcc aagataaatt
     2521 gtacccattc acttgggatg ctgtcagata caacggtaag cttatagcat atccaattgc
     2581 tgttgaagcc ttatcgttga tctacaataa ggacttgtta cctaacccac caaagacatg
     2641 ggaagaaatt ccagctttgg ataaagagct aaaggccaag ggaaaaagcg ctttgatgtt
     2701 taacttacaa gaaccttatt tcacctggcc attaatcgct gctgacggtg gttacgcatt
     2761 taaatacgaa aacggtaagt atgacattaa ggatgtcggc gttgataatg ctggtgcaaa
     2821 ggctggtttg accttcttgg ttgatttaat aaagaataaa cacatgaacg ctgacactga
     2881 ctactctatc gccgaagccg ctttcaacaa gggtgaaact gctatgacca taaatggtcc
     2941 atgggcctgg agtaacattg atacttctaa agtaaactat ggtgtcactg ttttgcctac
     3001 attcaaaggc caaccatcca agccctttgt cggtgttcta tccgctggca tcaatgctgc
     3061 ttctccaaac aaggagctag ctaaagaatt tttggaaaat tacttgttaa ctgacgaagg
     3121 tttggaggcc gttaacaagg acaagccatt gggtgctgtc gctttgaaat cttacgaaga
     3181 agaattggct aaggatccca gaattgccgc taccatggag aacgcccaaa agggtgaaat
     3241 tatgccaaac atcccacaaa tgtcagcctt ctggtacgca gttaggactg ccgtgattaa
     3301 tgctgcttcc ggtagacaaa ccgtcgacga agcattaaaa gatgctcaaa cgtaaacagg
     3361 ccccttttcc tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc
     3421 tcccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
     3481 ttattttttt taatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     3541 ttttttctgt acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga
     3601 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
     3661 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
     3721 acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca
     3781 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg
     3841 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg
     3901 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg
     3961 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat
     4021 tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac
     4081 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg
     4141 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga
     4201 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact
     4261 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa
     4321 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt
     4381 ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct
     4441 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag
     4501 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag
     4561 caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct
     4621 tcaatttaat tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga
     4681 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     4741 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt
     4801 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     4861 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct
     4921 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt
     4981 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag
     5041 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc
     5101 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat
     5161 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa
     5221 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt
     5281 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg
     5341 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat
     5401 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag
     5461 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt
     5521 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg
     5581 tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag
     5641 aataggaact tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca
     5701 atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct
     5761 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5821 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct
     5881 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5941 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6001 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6061 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6121 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6181 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6241 agcttccagg ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6301 ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6361 acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt
     6421 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     6481 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     6541 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     6601 tcacttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6661 ccatagttgc ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg
     6721 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6781 taaaccagcc agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca
     6841 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6901 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6961 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7021 aagcggttag ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat
     7081 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct
     7141 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7201 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7261 tgctcatcat tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga
     7321 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7381 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7441 cgacacggaa atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact
     7501 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     7561 gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt
     7621 atgatttaaa tggtcagtta ga
//
