LOCUS       p415TEF cyto roGFP(iL)-Grx1(human)        7775 bp ds-DNA     circular     24-MAR-2015
DEFINITION  sequence-optimized.
ACCESSION   . (modified)
VERSION     .
KEYWORDS    Untitled 2.
SOURCE      null
  ORGANISM  
            .
REFERENCE   1  (bases 1 to 7775)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dienstag, 24. 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..7775
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      4392..4639
                     /gene="S. cerevisiae CYC1"
                     /note="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     rep_origin      1746..2334
                     /direction="RIGHT"
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      4847..5302
                     /direction="RIGHT"
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             6019..7113
                     /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"
     CDS             4047..4364
                     /product="Grx1 (human)"
     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(4373..4389)
                     /note="KS primer"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4686..4702)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5602..6006
                     /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(4658..4676)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3951..4040
                     /product="Linker"
     CDS             3216..3944
                     /product="roGFP-iL"
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 tagcgaattc tctaaaggtg
     3241 aagaattgtt cacaggtgta gtccctattt tggtcgaatt ggatggtgac gtaaacggtc
     3301 ataagttctc agtatcaggt gaaggtgaag gtgacgcaac atacggtaaa ttgaccttga
     3361 agtttatttc aactactggt aaattgccag ttccttggcc aactttggta accactttaa
     3421 gttatggtgt acaatgtttc tcaagatacc ctgatcatat gaaacaacac gactttttca
     3481 agtctgctat gccagaaggt tacgttcaag aaagaacaat tttctttaag gatgacggta
     3541 actacaagac tagagcagaa gtcaaatttg aaggtgacac attggttaac agaatcgaat
     3601 tgaagggtat cgatttcaag gaagacggta acatcttggg tcataaattg gaatacaact
     3661 acaactgttt gtcaaacgta tacatcatgg ccgataagca aaagaatggt atcaaagtca
     3721 acttcaagat cagacataac atcgaagatg gttccgttca attagctgac cactatcaac
     3781 aaaatacccc tattggtgac ggtcctgttt tgttaccaga caaccattac ttgtctacat
     3841 gctcagcttt atccaaagat ccaaatgaaa agagagacca catggtcttg ttagaatttg
     3901 tcaccgccgc aggtatcaca ttgggtatgg acgaattgta taaaactagt ggtggttcag
     3961 gtggtggtgg ttcaggtggt ggtggttcag gtggaggagg atcaggagga ggaggatcag
     4021 gaggaggagg atcaggagga gaattcgctc aagagtttgt gaactgcaaa atccagcctg
     4081 ggaaggtggt tgtgttcatc aagcccacct gcccgtactg caggagggcc caagagatcc
     4141 tcagtcaatt gcccatcaaa caagggcttc tggaatttgt cgatatcaca gccaccaacc
     4201 acactaacga gattcaagat tatttgcaac agctcacggg agcaagaacg gtgcctcgag
     4261 tctttattgg taaagattgt ataggcggat gcagtgatct agtctctttg caacagagtg
     4321 gggaactgct gacgcggcta aagcagattg gagctctgca gtaaaagctt atcgataccg
     4381 tcgacctcga gtcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc
     4441 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     4501 tatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta
     4561 cagacgcgtg tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg
     4621 ctcgaaggct ttaatttgcg gccggtaccc aattcgccct atagtgagtc gtattacgcg
     4681 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     4741 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     4801 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcgacgc gccctgtagc
     4861 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4921 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4981 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     5041 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     5101 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5161 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     5221 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     5281 aaaatattaa cgtttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     5341 tttcacaccg catatcgacg gtcgaggaga acttctagta tatccacata cctaatatta
     5401 ttgccttatt aaaaatggaa tcccaacaat tacatcaaaa tccacattct cttcaaaatc
     5461 aattgtcctg tacttccttg ttcatgtgtg ttcaaaaacg ttatatttat aggataatta
     5521 tactctattt ctcaacaagt aattggttgt ttggccgagc ggtctaaggc gcctgattca
     5581 agaaatatct tgaccgcagt taactgtggg aatactcagg tatcgtaaga tgcaagagtt
     5641 cgaatctctt agcaaccatt atttttttcc tcaacataac gagaacacac aggggcgcta
     5701 tcgcacagaa tcaaattcga tgactggaaa ttttttgtta atttcagagg tcgcctgacg
     5761 catatacctt tttcaactga aaaattggga gaaaaaggaa aggtgagagg ccggaaccgg
     5821 cttttcatat agaatagaga agcgttcatg actaaatgct tgcatcacaa tacttgaagt
     5881 tgacaatatt atttaaggac ctattgtttt ttccaatagg tggttagcaa tcgtcttact
     5941 ttctaacttt tcttaccttt tacatttcag caatatatat atatatttca aggatatacc
     6001 attctaatgt ctgcccctat gtctgcccct aagaagatcg tcgttttgcc aggtgaccac
     6061 gttggtcaag aaatcacagc cgaagccatt aaggttctta aagctatttc tgatgttcgt
     6121 tccaatgtca agttcgattt cgaaaatcat ttaattggtg gtgctgctat cgatgctaca
     6181 ggtgtcccac ttccagatga ggcgctggaa gcctccaaga aggttgatgc cgttttgtta
     6241 ggtgctgtgg gtggtcctaa atggggtacc ggtagtgtta gacctgaaca aggtttacta
     6301 aaaatccgta aagaacttca attgtacgcc aacttaagac catgtaactt tgcatccgac
     6361 tctcttttag acttatctcc aatcaagcca caatttgcta aaggtactga cttcgttgtt
     6421 gtcagagaat tagtgggagg tatttacttt ggtaagagaa aggaagacga tggtgatggt
     6481 gtcgcttggg atagtgaaca atacaccgtt ccagaagtgc aaagaatcac aagaatggcc
     6541 gctttcatgg ccctacaaca tgagccacca ttgcctattt ggtccttgga taaagctaat
     6601 gttttggcct cttcaagatt atggagaaaa actgtggagg aaaccatcaa gaacgaattc
     6661 cctacattga aggttcaaca tcaattgatt gattctgccg ccatgatcct agttaagaac
     6721 ccaacccacc taaatggtat tataatcacc agcaacatgt ttggtgatat catctccgat
     6781 gaagcctccg ttatcccagg ttccttgggt ttgttgccat ctgcgtcctt ggcctctttg
     6841 ccagacaaga acaccgcatt tggtttgtac gaaccatgcc acggttctgc tccagatttg
     6901 ccaaagaata aggttgaccc tatcgccact atcttgtctg ctgcaatgat gttgaaattg
     6961 tcattgaact tgcctgaaga aggtaaggcc attgaagatg cagttaaaaa ggttttggat
     7021 gcaggtatca gaactggtga tttaggtggt tccaacagta ccaccgaagt cggtgatgct
     7081 gtcgccgaag aagttaagaa aatccttgct taaaaagatt ctcttttttt atgatatttg
     7141 tacataaact ttataaatga aattcataat agaaacgaca cgaaattaca aaatggaata
     7201 tgttcatagg gtagacgaaa ctatatacgc aatctacata catttatcaa gaaggagaaa
     7261 aaggaggata gtaaaggaat acaggtaagc aaattgatac taatggctca acgtgataag
     7321 gaaaaagaat tgcactttaa cattaatatt gacaaggagg agggcaccac acaaaaagtt
     7381 aggtgtaaca gaaaatcatg aaactacgat tcctaatttg atattggagg attttctcta
     7441 aaaaaaaaaa aatacaacaa ataaaaaaca ctcaatgacc tgaccatttg atggagttta
     7501 agtcaatacc ttcttgaacc atttcccata atggtgaaag ttccctcaag aattttactc
     7561 tgtcagaaac ggccttacga cgtagtcgat atggtgcact ctcagtacaa tctgctctga
     7621 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     7681 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     7741 tcagaggttt tcaccgtcat caccgaaacg cgcga
//
