LOCUS       40924_3204        8361 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pAAV-TRE-Syn-Fluc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8361)
  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 8361)
  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 8361)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8361)
  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..8361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   19..162
                     /label=ITR
                     /note="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"
     polyA_signal    616..664
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     promoter        739..1186
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             1195..2844
                     /label=luciferase
                     /note="firefly luciferase"
     misc_feature    2848..2901
                     /label=2A peptide from Thosea asigna virus
                     /note="2A peptide from Thosea asigna virus"
     CDS             2848..2901
                     /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             2914..3759
                     /label=tTS
                     /note="fusion of a hybrid tetracycline repressor TetR with
                     a nuclear localization signal and a KRAB silencing domain"
     misc_feature    3763..3816
                     /label=2A peptide from Thosea asigna virus
                     /note="2A peptide from Thosea asigna virus"
     CDS             3763..3816
                     /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             3829..4482
                     /label=hmKO
                     /note="humanized monomeric Kusabira-Orange fluorescent
                     protein"
     misc_feature    4518..5106
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    5148..5355
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5365..5508
                     /label=ITR
                     /note="ITR"
     rep_origin      5607..6062
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6394..6498
                     /label=AmpR promoter
     CDS             6499..7356
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      7530..8118
                     /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 gtacccgggg atcctctaga aatattaagg tacgggaggt
      601 acttggagcg gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg
      661 tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta
      721 gcaaaatagg ctgtccccag tgcaagtggg ttttaggacc aggatgaggc ggggtggggg
      781 tgcctacctg acgaccgacc ccgacccact ggacaagcac ccaaccccca ttccccaaat
      841 tgcgcatccc ctatcagaga gggggagggg aaacaggatg cggcgaggcg cgtgcgcact
      901 gccagcttca gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg
      961 cctcagcact gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc cccttcccgg
     1021 ccaccttggt cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga
     1081 gatagggggg cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag cgctgcctca
     1141 gtctgcggtg ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga attcatggaa
     1201 gacgccaaaa acataaagaa aggcccggcg ccattctatc cgctagagga tggaaccgct
     1261 ggagagcaac tgcataaggc tatgaagaga tacgccctgg ttcctggaac aattgctttt
     1321 acagatgcac atatcgaggt gaacatcacg tacgcggaat acttcgaaat gtccgttcgg
     1381 ttggcagaag ctatgaaacg atatgggctg aatacaaatc acagaatcgt cgtatgcagt
     1441 gaaaactctc ttcaattctt tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt
     1501 gcgcccgcga acgacattta taatgaacgt gaattgctca acagtatgaa catttcgcag
     1561 cctaccgtag tgtttgtttc caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaaa
     1621 ttaccaataa tccagaaaat tattatcatg gattctaaaa cggattacca gggatttcag
     1681 tcgatgtaca cgttcgtcac atctcatcta cctcccggtt ttaatgaata cgattttgta
     1741 ccagagtcct ttgatcgtga caaaacaatt gcactgataa tgaactcctc tggatctact
     1801 gggttaccta agggtgtggc ccttccgcat agaactgcct gcgtcagatt ctcgcatgcc
     1861 agagatccta tttttggcaa tcaaatcatt ccggatactg cgattttaag tgttgttcca
     1921 ttccatcacg gttttggaat gtttactaca ctcggatatt tgatatgtgg atttcgagtc
     1981 gtcttaatgt atagatttga agaagagctg tttttacgat cccttcagga ttacaaaatt
     2041 caaagtgcgt tgctagtacc aaccctattt tcattcttcg ccaaaagcac tctgattgac
     2101 aaatacgatt tatctaattt acacgaaatt gcttctgggg gcgcacctct ttcgaaagaa
     2161 gtcggggaag cggttgcaaa acgcttccat cttccaggga tacgacaagg atatgggctc
     2221 actgagacta catcagctat tctgattaca cccgaggggg atgataaacc gggcgcggtc
     2281 ggtaaagttg ttccattttt tgaagcgaag gttgtggatc tggataccgg gaaaacgctg
     2341 ggcgttaatc agagaggcga attatgtgtc agaggaccta tgattatgtc cggttatgta
     2401 aacaatccgg aagcgaccaa cgccttgatt gacaaggatg gatggctaca ttctggagac
     2461 atagcttact gggacgaaga cgaacacttc ttcatagttg accgcttgaa gtctttaatt
     2521 aaatacaaag gataccaggt ggcccccgct gaattggagt cgatattgtt acaacacccc
     2581 aacatcttcg acgcgggcgt ggcaggtctt cccgacgatg acgccggtga acttcccgcc
     2641 gccgttgttg ttttggagca cggaaagacg atgacggaaa aagagatcgt ggattacgtc
     2701 gccagtcaag taacaaccgc gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta
     2761 ccgaaaggtc ttaccggaaa actcgacgca agaaaaatca gagagatcct cataaaggcc
     2821 aagaagggcg gaaagtccaa attgctcgag ggcagaggaa gtcttctaac atgcggtgac
     2881 gtggaggaga atcccggccc tgcaccggga tccatgtcca gattagataa aagtaaagtg
     2941 attaacagcg cattagagct gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa
     3001 ctcgcccaga agctaggtgt agagcagcct acattgtatt ggcacgtgcg caacaagcag
     3061 actcttatga acatgctttc agaggcaata ctggcgaagc atcacacccg ttcagcaccg
     3121 ttaccgactg agagttggca gcagtttctc caggaaaatg ctctgagttt ccgtaaagca
     3181 ttactggtcc atcgtgatgg agcccgattg catataggga cctctcctac gcccccccag
     3241 tttgaacaag cagaggcgca actacgctgt ctatgcgatg cagggttttc ggtcgaggag
     3301 gctcttttca ttctgcaatc tatcagccat tttacgttgg gtgcagtatt agaggagcaa
     3361 gcaacaaacc agatagaaaa taatcatgtg atagacgctg caccaccatt attacaagag
     3421 gcatttaata ttcaggcgag aacctctgct gaaatggcct tccatttcgg gctgaaatca
     3481 ttaatatttg gattttctgc acagttagat gaaaaaaagc atacacccat tgaggatggt
     3541 aataaaccaa aaaagaagag aaagctagca gtgtcagtga catttgaaga tgtggctgtg
     3601 ctctttactc gggacgagtg gaagaagctg gatctgtctc agagaagcct gtaccgtgag
     3661 gtgatgctgg agaattacag caacctggcc tccatggcag gattcctgtt taccaaacca
     3721 aaggtgatct ccctgttgca gcaaggagag gacccgtatc tcgagggcag aggaagtctt
     3781 ctaacatgcg gtgacgtgga ggagaatccc ggccctgcac cgggatccat ggtgagtgtg
     3841 attaaaccag aaatgaagat gaggtactac atggacggct ccgtcaatgg gcatgagttc
     3901 acaattgaag gtgaaggcac aggcagacct tacgagggac atcaagagat gacactacgc
     3961 gtcacaatgg ccaagggcgg gccaatgcct ttcgcgtttg acttagtgtc acacgtgttc
     4021 tgttacggcc acagaccttt tactaaatat ccagaagaga taccagacta tttcaaacaa
     4081 gcatttcctg aaggcctgtc atgggaaagg tcgttggagt tcgaagatgg tgggtccgct
     4141 tcagtcagtg cgcatataag ccttagagga aacaccttct accacaaatc caaatttact
     4201 ggggttaact ttcctgccga tggtcctatc atgcaaaacc aaagtgttga ttgggagcca
     4261 tcaaccgaga aaattactgc cagcgacgga gttctgaagg gtgatgttac gatgtaccta
     4321 aaacttgaag gaggcggcaa tcacaaatgc caattcaaga ctacttacaa ggcggcaaaa
     4381 aagattctta aaatgccagg aagccattac atcagccatc gcctcgtcag gaaaaccgaa
     4441 ggcaacatta ctgagctggt agaagatgca gtagctcatt cctgatgtac aagtaaagcg
     4501 gccgcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     4561 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     4621 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     4681 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     4741 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     4801 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     4861 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     4921 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     4981 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     5041 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     5101 ccccgcctga tcgataccgt cgactagagc tcgctgatca gcctcgactg tgccttctag
     5161 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     5221 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     5281 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     5341 caggcatgct ggggagagat ctgaggaacc cctagtgatg gagttggcca ctccctctct
     5401 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     5461 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaacc cccccccccc
     5521 cccccctgca gcctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     5581 gcgtagcctg aatggcgaat ggcgcgacgc gccctgtagc ggcgcattaa gcgcggcggg
     5641 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     5701 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5761 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5821 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5881 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5941 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     6001 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     6061 ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     6121 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     6181 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     6241 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     6301 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     6361 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6421 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6481 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6541 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6601 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6661 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6721 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6781 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6841 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6901 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6961 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     7021 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     7081 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     7141 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     7201 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     7261 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     7321 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     7381 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7441 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7501 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7561 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7621 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     7681 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7741 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7801 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7861 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7921 cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7981 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     8041 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     8101 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     8161 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     8221 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     8281 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     8341 attcattaat gcaggctgca g
//
