LOCUS       Exported                8258 bp ds-DNA     circular SYN 19-AUG-2015
DEFINITION  Yeast/E.coli shuttle vector built from pRS416.  Expresses the Z3EV 
            transcription factor and contains a Z3 promoter element driving 
            yEGFP.
ACCESSION   .
VERSION     .
KEYWORDS    RB3579
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8258)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 19, 2015 from SnapGene 2.8.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8258
                     /organism="synthetic DNA construct"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
     gene            191..1291
                     /note="URA3"
                     /note="URA3 from S. cerevisiae; contains the full-length 
                     ORF  220 bp upstream and 78 bp downstream.  Notably there 
                     are a few mutations compared to the SGD sequence."
     gene            complement(1999..2715)
                     /note="yEGFP"
                     /note="Yeast codon-optimized E-GFP (sequence from the Kurt 
                     Thorn vector pKT127); i included AAC infront of the start 
                     codon so that there is an ""A"" at the -3 position for 
                     increased expression."
     promoter        complement(2725..3423)
                     /note="Z3pr"
                     /note="Z3 promoter (Z3EV binding sites have replaced the 
                     Gal4 binding sites in this modified GAL1 promoter)"
     misc_feature    3430..5450
                     /note="ACT1pr-yZ3EV-3'UTR"
                     /note="Yeast codon-optimized Z3EV transcription factor 
                     under the control of the ACT1 promoter; has the A. gossypii
                     TEF3 3'UTR"
     rep_origin      5892..6520
                     /note="Bluescript (ColE1) origin"
                     /note="The bacterial origin of replication from the 
                     bluescript vector (pBluescript) that was used as the 
                     original backbone for the pRS plasmids.  *This ColE1 origin
                     is modified from the original and supports higher plasmid 
                     copy number (300-500 copies per cell)."
     gene            complement(6669..7529)
                     /note="bla"
                     /note="bla encodes the ampicillin-resistance enzyme 
                     (beta-lactamase)... this casette also includes 198 bp 
                     upstream that contains the promoter."
     rep_origin      7661..8176
                     /note="CEN6/ARS4"
                     /note="Contains the autonomous replicating sequence 
                     (origin)  ARS4  and the centromeric sequence  CEN6  from S.
                     cerevisiae."
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc
      241 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
      301 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
      361 cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc
      421 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
      481 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
      541 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
      601 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
      661 caatttttta ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca
      721 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
      781 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
      841 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
      901 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
      961 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     1021 gggtttagat gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt
     1081 ctctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     1141 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
     1201 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
     1261 agcttcaatt taattatatc agttattacc ctatgcggtg tgaaataccg cacagatgcg
     1321 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     1381 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     1441 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     1501 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     1561 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     1621 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     1681 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     1741 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     1801 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     1861 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     1921 ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta
     1981 tagggcgaat tgggtacctt atttgtacaa ttcatcaata ccatgggtaa taccagcagc
     2041 agtaacaaat tctaacaaga ccatgtggtc tctcttttcg tttggatctt tggataaggc
     2101 agattgagtg gataagtaat ggttgtctgg taacaagact ggaccatcac caattggagt
     2161 attttgttga taatggtcag ctaattgaac agaaccatct tcaatgttgt gtctaatttt
     2221 gaagttaact ttgataccat tcttttgttt gtcagccatg atgtaaacat tgtgagagtt
     2281 atagttgtat tccaatttgt gacctaaaat gttaccatct tctttaaaat caataccttt
     2341 taattcgatt ctattaacta aggtatcacc ttcaaacttg acttcagctc tggtcttgta
     2401 gttaccgtca tctttgaaaa aaatagttct ttcttgaaca taaccttctg gcatggcaga
     2461 cttgaaaaag tcatgttgtt tcatatgatc tgggtatcta gaaaaacatt gaacaccata
     2521 agttaaagta gtgactaagg ttggccatgg aactggcaat ttaccagtag tacaaataaa
     2581 ttttaaggtc aatttaccgt aagtagcatc accttcacct tcaccggaga cagaaaattt
     2641 gtgaccatta acatcaccat ctaattcaac caaaattggg acaacaccag tgaataattc
     2701 ttcaccttta gacatgttct cgagtatagt tttttctcct tgacgttaaa gtatagaggt
     2761 atattaacaa ttttttgttg atacttttat gacatttgaa taagaagtaa tacaaactga
     2821 aaatgttgaa agtattagtt aaagtggtta tgcagctttt ccatttatat atctgttaat
     2881 agatcaaaaa tcatcgcttc gctgattaat taccccagaa ataaggctaa aaaactaatc
     2941 gcattatcat cctatggttg ttaatttgat tcgttaattt gaaggtttgt ggggccaggt
     3001 tactgccaat ttttcctctt cataaccata aaagctagta ttgtagaatc tttattgttc
     3061 ggagcagtgc ggcgcgaggc acatctgcgt ttcaggaacg cgaccggtga agacgaggac
     3121 gcacggtcta gacgcccacg cacgcccacg cccgcccacg cacgcccacg cccgcccacg
     3181 cacgcccacg cggccgcact tcaatatagc aatgagcagt taagcgtatt actgaaagtt
     3241 ccaaagagaa ggttttttta ggctaagata atggggctct ttacatttcc acaacatata
     3301 agtaagatta gatatggata tgtatatgga tatgtatatg gtggtaatgc catgtaatat
     3361 gattattaaa cttctttgcg tccatccaaa aaaaaagtaa gaatttttga aaattcaata
     3421 taaactagtc cctttaaaaa catatgcctc accctaacat attttccaat taaccctcaa
     3481 tatttctctg tcacccggcc tctattttcc attttcttct ttacccgcca cgcgtttttt
     3541 tctttcaaat ttttttcttc cttcttcttt ttcttccacg tcctcttgca taaataaata
     3601 aaccgttttg aaaccaaact cgcctctctc tctccttttt gaaatatttt tgggtttgtt
     3661 tgatcctttc cttcccaatc tctcttgttt aatatatatt catttatatc acgctctctt
     3721 tttatcttcc tttttttcct ctctcttgta ttcttccttc ccctttctac tcaaaccaag
     3781 aagaaaaaga aaaggtcaat ctttgttaaa gaataggatc ttctactaca tcagctttta
     3841 gatttttcac gcttactgct tttttcttcc caagatcgaa aatttactga attaacaatg
     3901 ggaacgagac cctacgcttg ccctgtagaa agttgtgata ggagattcag cagatctgac
     3961 gagctgacta ggcatattcg tattcacacc ggccaaaagc ctttccaatg tagaatctgt
     4021 atgagaaact tcagcagaag tgaccatttg actacgcaca ttagaaccca caccggcgag
     4081 aagccgttcg cgtgtgacat ttgtggaaga aagttcgcca gatctgatga aagaaagaga
     4141 catacaaaga tccatacagg aggtgggggc actccggcag cagcctcaac cctagaagat
     4201 ccaagcgctg gagacatgag ggcagccaac ttgtggccaa gcccattaat gatcaaaaga
     4261 tctaagaaga attctcttgc actgtctttg accgctgatc aaatggtttc cgcactactt
     4321 gatgctgaac cacctatatt gtacagtgaa tacgacccaa ctcgtccttt ttccgaggct
     4381 tccatgatgg gtcttttgac aaatctagca gacagggaat tagttcatat gattaattgg
     4441 gccaaaagag ttcccggctt tgttgacttg acattacatg accaggtaca tcttcttgaa
     4501 tgcgcgtggt tagaaatatt gatgatcgga ttagtgtgga gaagtatgga gcaccctgta
     4561 aaattattat ttgcacctaa tttattattg gacaggaacc aaggtaaatg cgtagaaggt
     4621 atggttgaga tttttgatat gcttcttgcc actagctcca ggttcagaat gatgaatttg
     4681 caaggagaag aattcgtttg cctgaaatca attattttgt taaatagcgg tgtctataca
     4741 tttttaagct caactctgaa aagcctggaa gagaaagacc atatacatag agtgttggac
     4801 aagatcactg atacattgat tcaccttatg gcgaaagcag gattgactct gcagcaacag
     4861 catcagaggc tagctcagtt actactaatt ttgtcacaca ttagacatat gtctaacaag
     4921 ggtatggagc acctgtattc tatgaagtgt aaaaacgtcg ttcctctgta tgatttatta
     4981 ctagaaatgc tggacgctca caggttgcat gcaccaactt ctagaggcgg agcctccgtt
     5041 gaagaaaccg accaatccca cctagcgaca gcagggtcta catctagcga actgcatttg
     5101 gacggtgaag acgttgccat ggcccacgct gacgctttag acgattttga tttagatatg
     5161 ttgggtgacg gggactctcc agggcctggc ttcactccac atgattcagc accttacggt
     5221 gcgctggata tggctgactt cgaatttgag cagatgttca ccgatgctct gggtattgat
     5281 gagtatggtg gttagtagta agctggttct tctttctgcc atttaattgt agacttgttt
     5341 tctatttaat atcactgtat cctgtagcgt ttgtatacat tccgtaatag aaggtccgat
     5401 actttacttt catatcagtc gcagtttgcc acttgagggg agatcgacgc gagctccagc
     5461 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     5521 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5581 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5641 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     5701 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5761 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5821 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5881 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5941 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6001 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6061 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6121 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6181 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6241 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6301 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6361 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6421 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6481 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6541 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6601 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6661 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6721 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6781 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6841 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6901 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6961 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7021 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7081 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7141 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7201 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7261 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7321 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7381 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7441 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7501 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7561 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7621 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ggtccttttc atcacgtgct
     7681 ataaaaataa ttataattta aattttttaa tataaatata taaattaaaa atagaaagta
     7741 aaaaaagaaa ttaaagaaaa aatagttttt gttttccgaa gatgtaaaag actctagggg
     7801 gatcgccaac aaatactacc ttttatcttg ctcttcctgc tctcaggtat taatgccgaa
     7861 ttgtttcatc ttgtctgtgt agaagaccac acacgaaaat cctgtgattt tacattttac
     7921 ttatcgttaa tcgaatgtat atctatttaa tctgcttttc ttgtctaata aatatatatg
     7981 taaagtacgc tttttgttga aattttttaa acctttgttt attttttttt cttcattccg
     8041 taactcttct accttcttta tttactttct aaaatccaaa tacaaaacat aaaaataaat
     8101 aaacacagag taaattccca aattattcca tcattaaaag atacgaggcg cgtgtaagtt
     8161 acaggcaagc gatccgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     8221 cgtatcacga ggccctttcg tcacgaggcc ctttcgtc
//
