LOCUS       Exported                5293 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 leader protein.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046449
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5293)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5293)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5293
                     /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             968..985
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             986..1003
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1048..1066)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1417..2220
                     /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      2303..3285
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3363..3392
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3414..4001
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      4013..4040
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4064..4255)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4256..5116)
                     /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     4879..4898
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5117..5219)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      5220..5251
                     /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 atggagagcc ttgtccctgg tttcaacgag aaaacacacg tccaactcag
      481 tttgcctgtt ttacaggttc gcgacgtgct cgtacgtggc tttggagact ccgtggagga
      541 ggtcttatca gaggcacgtc aacatcttaa agatggcact tgtggcttag tagaagttga
      601 aaaaggcgtt ttgcctcaac ttgaacagcc ctatgtgttc atcaaacgtt cggatgctcg
      661 aactgcacct catggtcatg ttatggttga gctggtagca gaactcgaag gcattcagta
      721 cggtcgtagt ggtgagacac ttggtgtcct tgtccctcat gtgggcgaaa taccagtggc
      781 ttaccgcaag gttcttcttc gtaagaacgg taataaagga gctggtggcc atagttacgg
      841 cgccgatcta aagtcatttg acttaggcga cgagcttggc actgatcctt atgaagattt
      901 tcaagaaaac tggaacacta aacatagcag tggtgttacc cgtgaactca tgcgtgagct
      961 taacggactc gtgccgagag gttctcacca tcatcaccat cattaaacag gccccttttc
     1021 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc
     1081 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     1141 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     1201 tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt
     1261 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
     1321 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
     1381 cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa
     1441 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag
     1501 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt
     1561 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt
     1621 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc
     1681 ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt
     1741 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt
     1801 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg
     1861 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact
     1921 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg
     1981 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac
     2041 aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct
     2101 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt
     2161 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
     2221 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
     2281 ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg
     2341 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     2401 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     2461 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     2521 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     2581 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     2641 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     2701 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     2761 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     2821 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     2881 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     2941 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     3001 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     3061 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     3121 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     3181 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag
     3241 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
     3301 ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca
     3361 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat
     3421 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3481 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag
     3541 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3601 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3661 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3721 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3781 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3841 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3901 gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc
     3961 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa
     4021 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt
     4081 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     4141 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     4201 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc
     4261 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     4321 cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg
     4381 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4441 cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta
     4501 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4561 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4621 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4681 gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg
     4741 ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga
     4801 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4861 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4921 ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt
     4981 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5041 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5101 aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     5161 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     5221 gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa
     5281 atggtcagtt aga
//
