LOCUS       40924_3199        8687 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pAAV-TRE-CAG-Fluc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8687)
  AUTHORS   Chen Y, Cao L, Luo C, Ditzel DA, Peter J, Sprengel R.
  TITLE     RANGE: Gene Transfer of Reversibly Controlled Polycistronic Genes
  JOURNAL   Mol Ther Nucleic Acids 2, E85 (2013)
  PUBMED    23571608
REFERENCE   2  (bases 1 to 8687)
  AUTHORS   Chen Y, Sprengel R.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-NOV-2012) Molecular Neurobiology, Max Planck institute
            for medical research, Jahnstr. 29, Heidelberg, BW 69120, Germany
REFERENCE   3  (bases 1 to 8687)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8687)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Ther 
            Nucleic Acids 2, E85 (2013)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-NOV-2012) Molecular Neurobiology, Max Planck institute for 
            medical research, Jahnstr. 29, Heidelberg, BW 69120, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   19..162
                     /label=AAV2 ITR
                     /note="AAV2 ITR"
     protein_bind    complement(258..528)
                     /label=tetracycline response element
                     /note="contains seven copies of the tetracycline operator
                     tetO"
     regulatory      543..559
                     /label=CMVmin
                     /note="CMVmin"
                     /regulatory_class="promoter"
     promoter        814..1091
                     /label=chicken beta-actin promoter
     CDS             1521..3170
                     /label=luciferase
                     /note="firefly luciferase"
     misc_feature    3174..3227
                     /label=2A peptide from Thosea asigna virus
                     /note="2A peptide from Thosea asigna virus"
     CDS             3174..3227
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3240..4085
                     /label=tTS
                     /note="fusion of a hybrid tetracycline repressor TetR with
                     a nuclear localization signal and a KRAB silencing domain"
     misc_feature    4089..4142
                     /label=2A peptide from Thosea asigna virus
                     /note="2A peptide from Thosea asigna virus"
     CDS             4089..4142
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4155..4808
                     /label=hmKO
                     /note="humanized monomeric Kusabira-Orange fluorescent
                     protein"
     misc_feature    4844..5432
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    5474..5681
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5691..5834
                     /label=AAV2 ITR
                     /note="AAV2 ITR"
     rep_origin      5933..6388
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6720..6824
                     /label=AmpR promoter
     CDS             6825..7682
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      7856..8444
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gggggggggg ggggggggtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
       61 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
      121 gcgcgcagag agggagtggc caactccatc actaggggtt cctagatcta ggatcacgcg
      181 ttctagtcct agggatatct tcgaaccatg gttaattaag tttaaacatc gatcaattcg
      241 agctcgactt tcacttttct ctatcactga tagggagtgg taaactcgac tttcactttt
      301 ctctatcact gatagggagt ggtaaactcg actttcactt ttctctatca ctgataggga
      361 gtggtaaact cgactttcac ttttctctat cactgatagg gagtggtaaa ctcgactttc
      421 acttttctct atcactgata gggagtggta aactcgactt tcacttttct ctatcactga
      481 tagggagtgg taaactcgac tttcactttt ctctatcact gatagggagt ggtaaactcg
      541 acctatataa gcagagctcg gtaccattga cgtcaataat gacgtatgtt cccatagtaa
      601 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      661 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      721 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      781 acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca
      841 ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat
      901 tttgtgcagc gatgggggcg gggggggggg gggggcgcgc gccaggcggg gcggggcggg
      961 gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc
     1021 tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg
     1081 cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct
     1141 cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg
     1201 gcccttctcc tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt
     1261 ggctgcgtga aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg
     1321 ggctgtccgc ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc
     1381 tggcgtgtga ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc
     1441 ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattg
     1501 gatctagtcg attcgaattc atggaagacg ccaaaaacat aaagaaaggc ccggcgccat
     1561 tctatccgct agaggatgga accgctggag agcaactgca taaggctatg aagagatacg
     1621 ccctggttcc tggaacaatt gcttttacag atgcacatat cgaggtgaac atcacgtacg
     1681 cggaatactt cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata
     1741 caaatcacag aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg
     1801 gcgcgttatt tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat
     1861 tgctcaacag tatgaacatt tcgcagccta ccgtagtgtt tgtttccaaa aaggggttgc
     1921 aaaaaatttt gaacgtgcaa aaaaaattac caataatcca gaaaattatt atcatggatt
     1981 ctaaaacgga ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc
     2041 ccggttttaa tgaatacgat tttgtaccag agtcctttga tcgtgacaaa acaattgcac
     2101 tgataatgaa ctcctctgga tctactgggt tacctaaggg tgtggccctt ccgcatagaa
     2161 ctgcctgcgt cagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg
     2221 atactgcgat tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg
     2281 gatatttgat atgtggattt cgagtcgtct taatgtatag atttgaagaa gagctgtttt
     2341 tacgatccct tcaggattac aaaattcaaa gtgcgttgct agtaccaacc ctattttcat
     2401 tcttcgccaa aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt
     2461 ctgggggcgc acctctttcg aaagaagtcg gggaagcggt tgcaaaacgc ttccatcttc
     2521 cagggatacg acaaggatat gggctcactg agactacatc agctattctg attacacccg
     2581 agggggatga taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg
     2641 tggatctgga taccgggaaa acgctgggcg ttaatcagag aggcgaatta tgtgtcagag
     2701 gacctatgat tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca
     2761 aggatggatg gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca
     2821 tagttgaccg cttgaagtct ttaattaaat acaaaggata ccaggtggcc cccgctgaat
     2881 tggagtcgat attgttacaa caccccaaca tcttcgacgc gggcgtggca ggtcttcccg
     2941 acgatgacgc cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga
     3001 cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg
     3061 gaggagttgt gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa
     3121 aaatcagaga gatcctcata aaggccaaga agggcggaaa gtccaaattg ctcgagggca
     3181 gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctgca ccgggatcca
     3241 tgtccagatt agataaaagt aaagtgatta acagcgcatt agagctgctt aatgaggtcg
     3301 gaatcgaagg tttaacaacc cgtaaactcg cccagaagct aggtgtagag cagcctacat
     3361 tgtattggca cgtgcgcaac aagcagactc ttatgaacat gctttcagag gcaatactgg
     3421 cgaagcatca cacccgttca gcaccgttac cgactgagag ttggcagcag tttctccagg
     3481 aaaatgctct gagtttccgt aaagcattac tggtccatcg tgatggagcc cgattgcata
     3541 tagggacctc tcctacgccc ccccagtttg aacaagcaga ggcgcaacta cgctgtctat
     3601 gcgatgcagg gttttcggtc gaggaggctc ttttcattct gcaatctatc agccatttta
     3661 cgttgggtgc agtattagag gagcaagcaa caaaccagat agaaaataat catgtgatag
     3721 acgctgcacc accattatta caagaggcat ttaatattca ggcgagaacc tctgctgaaa
     3781 tggccttcca tttcgggctg aaatcattaa tatttggatt ttctgcacag ttagatgaaa
     3841 aaaagcatac acccattgag gatggtaata aaccaaaaaa gaagagaaag ctagcagtgt
     3901 cagtgacatt tgaagatgtg gctgtgctct ttactcggga cgagtggaag aagctggatc
     3961 tgtctcagag aagcctgtac cgtgaggtga tgctggagaa ttacagcaac ctggcctcca
     4021 tggcaggatt cctgtttacc aaaccaaagg tgatctccct gttgcagcaa ggagaggacc
     4081 cgtatctcga gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc
     4141 ctgcaccggg atccatggtg agtgtgatta aaccagaaat gaagatgagg tactacatgg
     4201 acggctccgt caatgggcat gagttcacaa ttgaaggtga aggcacaggc agaccttacg
     4261 agggacatca agagatgaca ctacgcgtca caatggccaa gggcgggcca atgcctttcg
     4321 cgtttgactt agtgtcacac gtgttctgtt acggccacag accttttact aaatatccag
     4381 aagagatacc agactatttc aaacaagcat ttcctgaagg cctgtcatgg gaaaggtcgt
     4441 tggagttcga agatggtggg tccgcttcag tcagtgcgca tataagcctt agaggaaaca
     4501 ccttctacca caaatccaaa tttactgggg ttaactttcc tgccgatggt cctatcatgc
     4561 aaaaccaaag tgttgattgg gagccatcaa ccgagaaaat tactgccagc gacggagttc
     4621 tgaagggtga tgttacgatg tacctaaaac ttgaaggagg cggcaatcac aaatgccaat
     4681 tcaagactac ttacaaggcg gcaaaaaaga ttcttaaaat gccaggaagc cattacatca
     4741 gccatcgcct cgtcaggaaa accgaaggca acattactga gctggtagaa gatgcagtag
     4801 ctcattcctg atgtacaagt aaagcggccg caagcttatc gataatcaac ctctggatta
     4861 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     4921 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     4981 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     5041 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     5101 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     5161 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     5221 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     5281 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     5341 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     5401 gagtcggatc tccctttggg ccgcctcccc gcctgatcga taccgtcgac tagagctcgc
     5461 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     5521 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     5581 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     5641 aagggggagg attgggaaga caatagcagg catgctgggg agagatctga ggaaccccta
     5701 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgcccgggca
     5761 aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
     5821 gagggagtgg ccaacccccc cccccccccc cctgcagcct ggcgtaatag cgaagaggcc
     5881 cgcaccgatc gcccttccca acagttgcgt agcctgaatg gcgaatggcg cgacgcgccc
     5941 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     6001 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     6061 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     6121 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     6181 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     6241 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     6301 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     6361 tttaacaaaa tattaacgtt tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     6421 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     6481 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     6541 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     6601 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     6661 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     6721 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     6781 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     6841 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     6901 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     6961 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     7021 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     7081 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     7141 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     7201 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     7261 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     7321 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     7381 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     7441 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     7501 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     7561 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     7621 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     7681 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     7741 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7801 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7861 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7921 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7981 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     8041 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     8101 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     8161 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     8221 ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa
     8281 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     8341 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     8401 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     8461 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     8521 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     8581 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     8641 aaccgcctct ccccgcgcgt tggccgattc attaatgcag gctgcag
//
