LOCUS       p415 TEF cyto roGFP2-Grx2(yeast)        7781 bp ds-DNA     linear       23-MAR-2015
DEFINITION  sequence-optimized, yeast Grx2.
ACCESSION   . (modified)
VERSION     .
KEYWORDS    Untitled 17.
SOURCE      null
  ORGANISM  
            .
REFERENCE   1  (bases 1 to 7781)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Montag, 23. Mrz 2015 from SnapGene Viewer 2.6.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     misc_feature    70..573
                     /note="CEN/ARS"
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     protein_bind    2696..2712
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     source          1..7781
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      4398..4645
                     /gene="S. cerevisiae CYC1"
                     /note="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     CDS             6025..7119
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /note="LEU2"
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENH
                     LIGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQL
                     YANLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSE
                     QYTVPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLK
                     VQHQLIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPD
                     KNTAFGLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLD
                     AGIRTGDLGGSNSTTEVGDAVAEEVKKILA"
     CDS             715..1575
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     rep_origin      1746..2334
                     /direction="RIGHT"
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      4853..5308
                     /direction="RIGHT"
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     2720..2736
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3195..3211
                     /note="SK primer"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4379..4395)
                     /note="KS primer"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4692..4708)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5608..6012
                     /gene="S. cerevisiae LEU2"
                     /note="LEU2 promoter"
     promoter        2793..3191
                     /gene="S. cerevisiae TEF1"
                     /note="TEF1 promoter"
                     /note="promoter for EF-1-alpha"
     promoter        610..714
                     /gene="bla"
                     /note="AmpR promoter"
     promoter        2658..2688
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     promoter        2757..2775
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(4664..4682)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3945..4034
                     /product="Linker"
     CDS             4041..4370
                     /product="Grx2(yeast)"
     CDS             3216..3938
                     /product="roGFP2"
ORIGIN      
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     2761 aaccctcact aaagggaaca aaagctggag ctcatagctt caaaatgttt ctactccttt
     2821 tttactcttc cagattttct cggactccgc gcatcgccgt accacttcaa aacacccaag
     2881 cacagcatac taaatttccc ctctttcttc ctctagggtg tcgttaatta cccgtactaa
     2941 aggtttggaa aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa aaaggcaata
     3001 aaaattttta tcacgtttct ttttcttgaa aatttttttt ttgatttttt tctctttcga
     3061 tgacctccca ttgatattta agttaataaa cggtcttcaa tttctcaagt ttcagtttca
     3121 tttttcttgt tctattacaa ctttttttac ttcttgctca ttagaaagaa agcatagcaa
     3181 tctaatctaa gttttctaga actagtggat ccaccatggc tagcgagttc tcaaagggtg
     3241 aagaattgtt tacaggtgtt gttcctattt tagtcgaatt ggacggtgac gttaatggtc
     3301 ataagtttag tgttagtggt gaaggtgaag gtgacgcaac atacggtaaa ttgaccttga
     3361 agtttatttc aactactggt aaattgccag ttccttggcc aactttggta accactttaa
     3421 catatggtgt acaatgtttc agtagatacc ctgatcatat gaaacaacac gactttttca
     3481 agtctgctat gccagaaggt tacgttcaag aaagaactat tttctttaag gatgacggta
     3541 actacaagac cagagcagaa gtcaaatttg aaggtgacac tttggttaac agaatcgaat
     3601 tgaagggtat cgatttcaag gaagacggta acatcttggg tcataaattg gaatacaact
     3661 acaactgtca caatgtatac ataatggccg ataagcaaaa gaatggtatc aaagtcaact
     3721 tcaagatcag acataacatc gaagatggtt ctgttcaatt agctgaccac tatcaacaaa
     3781 atacacctat tggtgacggt cctgttttgt taccagacaa ccattacttg tctacctgct
     3841 cagctttatc caaagatcca aatgaaaaga gagaccatat ggtattgtta gaatttgtca
     3901 ccgcagcagg tatcacattg ggtatggatg aattgtatac tagtggtggt tcaggtggtg
     3961 gtggttcagg tggtggtggt tcaggtggag gaggatcagg aggaggagga tcaggaggag
     4021 gaggatcagg aggagaattc atggtatccc aggaaacagt tgctcacgta aaggatctga
     4081 ttggccaaaa ggaagtgttt gttgcagcaa agacatactg cccttactgt aaagctactt
     4141 tgtctaccct cttccaagaa ttgaacgttc ccaaatccaa ggcccttgtg ttggaattag
     4201 atgaaatgag caatggctca gagattcaag acgctttaga agaaatctcg ggccaaaaaa
     4261 ctgtacctaa cgtatacatc aatggcaagc acattggtgg taacagcgat ttggaaactt
     4321 tgaagaaaaa tggcaagtta gctgaaatat tgaagccggt atttcaatag aagcttatcg
     4381 ataccgtcga cctcgagtca tgtaattagt tatgtcacgc ttacattcac gccctccccc
     4441 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta
     4501 tttttttata gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt
     4561 tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt
     4621 gggacgctcg aaggctttaa tttgcggccg gtacccaatt cgccctatag tgagtcgtat
     4681 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     4741 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     4801 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cgacgcgccc
     4861 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4921 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4981 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     5041 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     5101 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5161 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     5221 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5281 tttaacaaaa tattaacgtt tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     5341 gcggtatttc acaccgcata tcgacggtcg aggagaactt ctagtatatc cacataccta
     5401 atattattgc cttattaaaa atggaatccc aacaattaca tcaaaatcca cattctcttc
     5461 aaaatcaatt gtcctgtact tccttgttca tgtgtgttca aaaacgttat atttatagga
     5521 taattatact ctatttctca acaagtaatt ggttgtttgg ccgagcggtc taaggcgcct
     5581 gattcaagaa atatcttgac cgcagttaac tgtgggaata ctcaggtatc gtaagatgca
     5641 agagttcgaa tctcttagca accattattt ttttcctcaa cataacgaga acacacaggg
     5701 gcgctatcgc acagaatcaa attcgatgac tggaaatttt ttgttaattt cagaggtcgc
     5761 ctgacgcata tacctttttc aactgaaaaa ttgggagaaa aaggaaaggt gagaggccgg
     5821 aaccggcttt tcatatagaa tagagaagcg ttcatgacta aatgcttgca tcacaatact
     5881 tgaagttgac aatattattt aaggacctat tgttttttcc aataggtggt tagcaatcgt
     5941 cttactttct aacttttctt accttttaca tttcagcaat atatatatat atttcaagga
     6001 tataccattc taatgtctgc ccctatgtct gcccctaaga agatcgtcgt tttgccaggt
     6061 gaccacgttg gtcaagaaat cacagccgaa gccattaagg ttcttaaagc tatttctgat
     6121 gttcgttcca atgtcaagtt cgatttcgaa aatcatttaa ttggtggtgc tgctatcgat
     6181 gctacaggtg tcccacttcc agatgaggcg ctggaagcct ccaagaaggt tgatgccgtt
     6241 ttgttaggtg ctgtgggtgg tcctaaatgg ggtaccggta gtgttagacc tgaacaaggt
     6301 ttactaaaaa tccgtaaaga acttcaattg tacgccaact taagaccatg taactttgca
     6361 tccgactctc ttttagactt atctccaatc aagccacaat ttgctaaagg tactgacttc
     6421 gttgttgtca gagaattagt gggaggtatt tactttggta agagaaagga agacgatggt
     6481 gatggtgtcg cttgggatag tgaacaatac accgttccag aagtgcaaag aatcacaaga
     6541 atggccgctt tcatggccct acaacatgag ccaccattgc ctatttggtc cttggataaa
     6601 gctaatgttt tggcctcttc aagattatgg agaaaaactg tggaggaaac catcaagaac
     6661 gaattcccta cattgaaggt tcaacatcaa ttgattgatt ctgccgccat gatcctagtt
     6721 aagaacccaa cccacctaaa tggtattata atcaccagca acatgtttgg tgatatcatc
     6781 tccgatgaag cctccgttat cccaggttcc ttgggtttgt tgccatctgc gtccttggcc
     6841 tctttgccag acaagaacac cgcatttggt ttgtacgaac catgccacgg ttctgctcca
     6901 gatttgccaa agaataaggt tgaccctatc gccactatct tgtctgctgc aatgatgttg
     6961 aaattgtcat tgaacttgcc tgaagaaggt aaggccattg aagatgcagt taaaaaggtt
     7021 ttggatgcag gtatcagaac tggtgattta ggtggttcca acagtaccac cgaagtcggt
     7081 gatgctgtcg ccgaagaagt taagaaaatc cttgcttaaa aagattctct ttttttatga
     7141 tatttgtaca taaactttat aaatgaaatt cataatagaa acgacacgaa attacaaaat
     7201 ggaatatgtt catagggtag acgaaactat atacgcaatc tacatacatt tatcaagaag
     7261 gagaaaaagg aggatagtaa aggaatacag gtaagcaaat tgatactaat ggctcaacgt
     7321 gataaggaaa aagaattgca ctttaacatt aatattgaca aggaggaggg caccacacaa
     7381 aaagttaggt gtaacagaaa atcatgaaac tacgattcct aatttgatat tggaggattt
     7441 tctctaaaaa aaaaaaaata caacaaataa aaaacactca atgacctgac catttgatgg
     7501 agtttaagtc aataccttct tgaaccattt cccataatgg tgaaagttcc ctcaagaatt
     7561 ttactctgtc agaaacggcc ttacgacgta gtcgatatgg tgcactctca gtacaatctg
     7621 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     7681 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     7741 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg a
//
