LOCUS       40924_3209        7993 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAAV-tTS, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7993)
  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 7993)
  AUTHORS   Chen Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-MAY-2012) Molecular Neurobiology, Max Planck Institute
            for Medical Research, Jahnstr. 29, Heidelberg, BW 69120, Germany
REFERENCE   3  (bases 1 to 7993)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7993)
  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 
            (01-MAY-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..7993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   19..163
                     /rpt_family="AAV2 ITR"
     polyA_signal    248..296
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     promoter        371..818
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             827..2476
                     /label=luciferase
                     /note="firefly luciferase"
     mat_peptide     2480..2533
                     /product="2A peptide"
                     /note="self-cleaving"
     CDS             2480..2533
                     /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             2546..3391
                     /label=tTS
                     /note="fusion of a hybrid tetracycline repressor TetR with
                     a nuclear localization signal and a KRAB silencing domain"
     mat_peptide     3395..3448
                     /product="2A peptide"
     CDS             3395..3448
                     /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             3461..4114
                     /label=hmKO
                     /note="humanized monomeric Kusabira-Orange fluorescent
                     protein"
     misc_feature    4150..4738
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    4780..4987
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4996..5140
                     /rpt_family="AAV2 ITR"
     rep_origin      5239..5694
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6026..6130
                     /label=AmpR promoter
     CDS             6131..6988
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      7162..7750
                     /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 agggatccgg ggatcctcta gaaatattaa ggtacgggag gtacttggag
      241 cggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc
      301 gatagtacta acatacgctc tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata
      361 ggctgtcccc agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc
      421 tgacgaccga ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc
      481 ccctatcaga gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt
      541 cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca
      601 ctgaaggcgc gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg
      661 gtcgcgtccg cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg
      721 ggcacgggcg cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg
      781 tgggcagcgg aggagtcgtg tcgtgcctga gagcgcagtc gaattcatgg aagacgccaa
      841 aaacataaag aaaggcccgg cgccattcta tccgctagag gatggaaccg ctggagagca
      901 actgcataag gctatgaaga gatacgccct ggttcctgga acaattgctt ttacagatgc
      961 acatatcgag gtgaacatca cgtacgcgga atacttcgaa atgtccgttc ggttggcaga
     1021 agctatgaaa cgatatgggc tgaatacaaa tcacagaatc gtcgtatgca gtgaaaactc
     1081 tcttcaattc tttatgccgg tgttgggcgc gttatttatc ggagttgcag ttgcgcccgc
     1141 gaacgacatt tataatgaac gtgaattgct caacagtatg aacatttcgc agcctaccgt
     1201 agtgtttgtt tccaaaaagg ggttgcaaaa aattttgaac gtgcaaaaaa aattaccaat
     1261 aatccagaaa attattatca tggattctaa aacggattac cagggatttc agtcgatgta
     1321 cacgttcgtc acatctcatc tacctcccgg ttttaatgaa tacgattttg taccagagtc
     1381 ctttgatcgt gacaaaacaa ttgcactgat aatgaactcc tctggatcta ctgggttacc
     1441 taagggtgtg gcccttccgc atagaactgc ctgcgtcaga ttctcgcatg ccagagatcc
     1501 tatttttggc aatcaaatca ttccggatac tgcgatttta agtgttgttc cattccatca
     1561 cggttttgga atgtttacta cactcggata tttgatatgt ggatttcgag tcgtcttaat
     1621 gtatagattt gaagaagagc tgtttttacg atcccttcag gattacaaaa ttcaaagtgc
     1681 gttgctagta ccaaccctat tttcattctt cgccaaaagc actctgattg acaaatacga
     1741 tttatctaat ttacacgaaa ttgcttctgg gggcgcacct ctttcgaaag aagtcgggga
     1801 agcggttgca aaacgcttcc atcttccagg gatacgacaa ggatatgggc tcactgagac
     1861 tacatcagct attctgatta cacccgaggg ggatgataaa ccgggcgcgg tcggtaaagt
     1921 tgttccattt tttgaagcga aggttgtgga tctggatacc gggaaaacgc tgggcgttaa
     1981 tcagagaggc gaattatgtg tcagaggacc tatgattatg tccggttatg taaacaatcc
     2041 ggaagcgacc aacgccttga ttgacaagga tggatggcta cattctggag acatagctta
     2101 ctgggacgaa gacgaacact tcttcatagt tgaccgcttg aagtctttaa ttaaatacaa
     2161 aggataccag gtggcccccg ctgaattgga gtcgatattg ttacaacacc ccaacatctt
     2221 cgacgcgggc gtggcaggtc ttcccgacga tgacgccggt gaacttcccg ccgccgttgt
     2281 tgttttggag cacggaaaga cgatgacgga aaaagagatc gtggattacg tcgccagtca
     2341 agtaacaacc gcgaaaaagt tgcgcggagg agttgtgttt gtggacgaag taccgaaagg
     2401 tcttaccgga aaactcgacg caagaaaaat cagagagatc ctcataaagg ccaagaaggg
     2461 cggaaagtcc aaattgctcg agggcagagg aagtcttcta acatgcggtg acgtggagga
     2521 gaatcccggc cctgcaccgg gatccatgtc cagattagat aaaagtaaag tgattaacag
     2581 cgcattagag ctgcttaatg aggtcggaat cgaaggttta acaacccgta aactcgccca
     2641 gaagctaggt gtagagcagc ctacattgta ttggcacgtg cgcaacaagc agactcttat
     2701 gaacatgctt tcagaggcaa tactggcgaa gcatcacacc cgttcagcac cgttaccgac
     2761 tgagagttgg cagcagtttc tccaggaaaa tgctctgagt ttccgtaaag cattactggt
     2821 ccatcgtgat ggagcccgat tgcatatagg gacctctcct acgccccccc agtttgaaca
     2881 agcagaggcg caactacgct gtctatgcga tgcagggttt tcggtcgagg aggctctttt
     2941 cattctgcaa tctatcagcc attttacgtt gggtgcagta ttagaggagc aagcaacaaa
     3001 ccagatagaa aataatcatg tgatagacgc tgcaccacca ttattacaag aggcatttaa
     3061 tattcaggcg agaacctctg ctgaaatggc cttccatttc gggctgaaat cattaatatt
     3121 tggattttct gcacagttag atgaaaaaaa gcatacaccc attgaggatg gtaataaacc
     3181 aaaaaagaag agaaagctag cagtgtcagt gacatttgaa gatgtggctg tgctctttac
     3241 tcgggacgag tggaagaagc tggatctgtc tcagagaagc ctgtaccgtg aggtgatgct
     3301 ggagaattac agcaacctgg cctccatggc aggattcctg tttaccaaac caaaggtgat
     3361 ctccctgttg cagcaaggag aggacccgta tctcgagggc agaggaagtc ttctaacatg
     3421 cggtgacgtg gaggagaatc ccggccctgc accgggatcc atggtgagtg tgattaaacc
     3481 agaaatgaag atgaggtact acatggacgg ctccgtcaat gggcatgagt tcacaattga
     3541 aggtgaaggc acaggcagac cttacgaggg acatcaagag atgacactac gcgtcacaat
     3601 ggccaagggc gggccaatgc ctttcgcgtt tgacttagtg tcacacgtgt tctgttacgg
     3661 ccacagacct tttactaaat atccagaaga gataccagac tatttcaaac aagcatttcc
     3721 tgaaggcctg tcatgggaaa ggtcgttgga gttcgaagat ggtgggtccg cttcagtcag
     3781 tgcgcatata agccttagag gaaacacctt ctaccacaaa tccaaattta ctggggttaa
     3841 ctttcctgcc gatggtccta tcatgcaaaa ccaaagtgtt gattgggagc catcaaccga
     3901 gaaaattact gccagcgacg gagttctgaa gggtgatgtt acgatgtacc taaaacttga
     3961 aggaggcggc aatcacaaat gccaattcaa gactacttac aaggcggcaa aaaagattct
     4021 taaaatgcca ggaagccatt acatcagcca tcgcctcgtc aggaaaaccg aaggcaacat
     4081 tactgagctg gtagaagatg cagtagctca ttcctgatgt acaagtaaag cggccgcaag
     4141 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     4201 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     4261 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     4321 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     4381 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     4441 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     4501 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     4561 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     4621 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     4681 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     4741 gatcgatacc gtcgactaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     4801 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     4861 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     4921 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     4981 ctggggagag atctgaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     5041 gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg
     5101 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa cccccccccc cccccccctg
     5161 cagcctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgtagcc
     5221 tgaatggcga atggcgcgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     5281 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     5341 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     5401 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     5461 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     5521 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     5581 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     5641 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttcctgat
     5701 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     5761 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     5821 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5881 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     5941 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     6001 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     6061 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     6181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     6241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     6301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     6361 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     6421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     6481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     6541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     6601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6661 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     7021 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     7081 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7141 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7201 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7261 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     7321 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7381 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7441 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7501 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga
     7561 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7621 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7681 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7741 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7801 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     7861 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     7921 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     7981 atgcaggctg cag
//
