LOCUS       p415 TEF cyto roGFP2-Grx1(yeast)        7790 bp ds-DNA     circular     23-MAR-2015
DEFINITION  sequence-optimized, yeast Grx1.
ACCESSION   . (modified)
VERSION     .
KEYWORDS    Untitled 16.
SOURCE      null
  ORGANISM  
            .
REFERENCE   1  (bases 1 to 7790)
  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..7790
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      4407..4654
                     /gene="S. cerevisiae CYC1"
                     /note="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     CDS             6034..7128
                     /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      4862..5317
                     /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(4388..4404)
                     /note="KS primer"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4701..4717)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5617..6021
                     /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(4673..4691)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3948..4037
                     /product="Linker"
     CDS             4047..4379
                     /product="Grx1 (yeast)"
     CDS             3216..3941
                     /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 aattgtataa aactagtggt ggttcaggtg
     3961 gtggtggttc aggtggtggt ggttcaggtg gaggaggatc aggaggagga ggatcaggag
     4021 gaggaggatc aggaggagaa ttcaccatgg tatctcaaga aactatcaag cacgtcaagg
     4081 accttattgc agaaaacgag atcttcgtcg catccaaaac gtactgtcca tactgccatg
     4141 cagccctaaa cacgcttttt gaaaagttaa aggttcccag gtccaaagtt ctggttttgc
     4201 aattgaatga catgaaggaa ggcgcagaca ttcaggctgc gttatatgag attaatggcc
     4261 aaagaaccgt gccaaacatc tatattaatg gtaaacatat tggaggcaac gacgacttgc
     4321 aggaattgag ggagactggt gaattggagg aattgttaga acctattctt gcaaattaaa
     4381 agcttatcga taccgtcgac ctcgagtcat gtaattagtt atgtcacgct tacattcacg
     4441 ccctcccccc acatccgctc taaccgaaaa ggaaggagtt agacaacctg aagtctaggt
     4501 ccctatttat ttttttatag ttatgttagt attaagaacg ttatttatat ttcaaatttt
     4561 tctttttttt ctgtacagac gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga
     4621 gaaggttttg ggacgctcga aggctttaat ttgcggccgg tacccaattc gccctatagt
     4681 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     4741 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     4801 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     4861 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4921 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4981 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     5041 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     5101 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     5161 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     5221 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     5281 aacgcgaatt ttaacaaaat attaacgttt acaatttcct gatgcggtat tttctcctta
     5341 cgcatctgtg cggtatttca caccgcatat cgacggtcga ggagaacttc tagtatatcc
     5401 acatacctaa tattattgcc ttattaaaaa tggaatccca acaattacat caaaatccac
     5461 attctcttca aaatcaattg tcctgtactt ccttgttcat gtgtgttcaa aaacgttata
     5521 tttataggat aattatactc tatttctcaa caagtaattg gttgtttggc cgagcggtct
     5581 aaggcgcctg attcaagaaa tatcttgacc gcagttaact gtgggaatac tcaggtatcg
     5641 taagatgcaa gagttcgaat ctcttagcaa ccattatttt tttcctcaac ataacgagaa
     5701 cacacagggg cgctatcgca cagaatcaaa ttcgatgact ggaaattttt tgttaatttc
     5761 agaggtcgcc tgacgcatat acctttttca actgaaaaat tgggagaaaa aggaaaggtg
     5821 agaggccgga accggctttt catatagaat agagaagcgt tcatgactaa atgcttgcat
     5881 cacaatactt gaagttgaca atattattta aggacctatt gttttttcca ataggtggtt
     5941 agcaatcgtc ttactttcta acttttctta ccttttacat ttcagcaata tatatatata
     6001 tttcaaggat ataccattct aatgtctgcc cctatgtctg cccctaagaa gatcgtcgtt
     6061 ttgccaggtg accacgttgg tcaagaaatc acagccgaag ccattaaggt tcttaaagct
     6121 atttctgatg ttcgttccaa tgtcaagttc gatttcgaaa atcatttaat tggtggtgct
     6181 gctatcgatg ctacaggtgt cccacttcca gatgaggcgc tggaagcctc caagaaggtt
     6241 gatgccgttt tgttaggtgc tgtgggtggt cctaaatggg gtaccggtag tgttagacct
     6301 gaacaaggtt tactaaaaat ccgtaaagaa cttcaattgt acgccaactt aagaccatgt
     6361 aactttgcat ccgactctct tttagactta tctccaatca agccacaatt tgctaaaggt
     6421 actgacttcg ttgttgtcag agaattagtg ggaggtattt actttggtaa gagaaaggaa
     6481 gacgatggtg atggtgtcgc ttgggatagt gaacaataca ccgttccaga agtgcaaaga
     6541 atcacaagaa tggccgcttt catggcccta caacatgagc caccattgcc tatttggtcc
     6601 ttggataaag ctaatgtttt ggcctcttca agattatgga gaaaaactgt ggaggaaacc
     6661 atcaagaacg aattccctac attgaaggtt caacatcaat tgattgattc tgccgccatg
     6721 atcctagtta agaacccaac ccacctaaat ggtattataa tcaccagcaa catgtttggt
     6781 gatatcatct ccgatgaagc ctccgttatc ccaggttcct tgggtttgtt gccatctgcg
     6841 tccttggcct ctttgccaga caagaacacc gcatttggtt tgtacgaacc atgccacggt
     6901 tctgctccag atttgccaaa gaataaggtt gaccctatcg ccactatctt gtctgctgca
     6961 atgatgttga aattgtcatt gaacttgcct gaagaaggta aggccattga agatgcagtt
     7021 aaaaaggttt tggatgcagg tatcagaact ggtgatttag gtggttccaa cagtaccacc
     7081 gaagtcggtg atgctgtcgc cgaagaagtt aagaaaatcc ttgcttaaaa agattctctt
     7141 tttttatgat atttgtacat aaactttata aatgaaattc ataatagaaa cgacacgaaa
     7201 ttacaaaatg gaatatgttc atagggtaga cgaaactata tacgcaatct acatacattt
     7261 atcaagaagg agaaaaagga ggatagtaaa ggaatacagg taagcaaatt gatactaatg
     7321 gctcaacgtg ataaggaaaa agaattgcac tttaacatta atattgacaa ggaggagggc
     7381 accacacaaa aagttaggtg taacagaaaa tcatgaaact acgattccta atttgatatt
     7441 ggaggatttt ctctaaaaaa aaaaaaatac aacaaataaa aaacactcaa tgacctgacc
     7501 atttgatgga gtttaagtca ataccttctt gaaccatttc ccataatggt gaaagttccc
     7561 tcaagaattt tactctgtca gaaacggcct tacgacgtag tcgatatggt gcactctcag
     7621 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     7681 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     7741 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
//
