LOCUS       40924_3027        8323 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pAAV-CAG-Fluc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8323)
  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 8323)
  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 8323)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8323)
  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..8323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   19..162
                     /label=AAV2 ITR
                     /note="AAV2 ITR"
     promoter        450..727
                     /label=chicken beta-actin promoter
     CDS             1157..2806
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     misc_feature    2810..2863
                     /label=2A peptide from Thosea asigna virus
                     /note="2A peptide from Thosea asigna virus"
     CDS             2810..2863
                     /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             2876..3721
                     /codon_start=1
                     /label=tTS
                     /note="fusion of a hybrid tetracycline repressor TetR with
                     a nuclear localization signal and a KRAB silencing domain"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     RNKQTLMNMLSEAILAKHHTRSAPLPTESWQQFLQENALSFRKALLVHRDGARLHIGTS
                     PTPPQFEQAEAQLRCLCDAGFSVEEALFILQSISHFTLGAVLEEQATNQIENNHVIDAA
                     PPLLQEAFNIQARTSAEMAFHFGLKSLIFGFSAQLDEKKHTPIEDGNKPKKKRKLAVSV
                     TFEDVAVLFTRDEWKKLDLSQRSLYREVMLENYSNLASMAGFLFTKPKVISLLQQGEDP
                     Y"
     misc_feature    3725..3778
                     /label=2A peptide from Thosea asigna virus
                     /note="2A peptide from Thosea asigna virus"
     CDS             3725..3778
                     /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             3791..4444
                     /codon_start=1
                     /label=hmKO
                     /note="humanized monomeric Kusabira-Orange fluorescent
                     protein"
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAKGGPMPFAFDLVSHVFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKKILKMPGSHYISHRLVRKTEGNITELVEDAVAHS"
     misc_feature    4480..5068
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    5110..5317
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5327..5470
                     /label=AAV2 ITR
                     /note="AAV2 ITR"
     rep_origin      5569..6024
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6356..6460
                     /label=AmpR promoter
     CDS             6461..7318
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7492..8080
                     /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 agggatgtac cattgacgtc aataatgacg tatgttccca tagtaacgcc
      241 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      301 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      361 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      421 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
      481 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      541 tgcagcgatg ggggcggggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga
      601 ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg
      661 aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg
      721 gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg
      781 ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc
      841 ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct
      901 gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcggggct
      961 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     1021 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
     1081 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattggatc
     1141 tagtcgattc gaattcatgg aagacgccaa aaacataaag aaaggcccgg cgccattcta
     1201 tccgctagag gatggaaccg ctggagagca actgcataag gctatgaaga gatacgccct
     1261 ggttcctgga acaattgctt ttacagatgc acatatcgag gtgaacatca cgtacgcgga
     1321 atacttcgaa atgtccgttc ggttggcaga agctatgaaa cgatatgggc tgaatacaaa
     1381 tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc
     1441 gttatttatc ggagttgcag ttgcgcccgc gaacgacatt tataatgaac gtgaattgct
     1501 caacagtatg aacatttcgc agcctaccgt agtgtttgtt tccaaaaagg ggttgcaaaa
     1561 aattttgaac gtgcaaaaaa aattaccaat aatccagaaa attattatca tggattctaa
     1621 aacggattac cagggatttc agtcgatgta cacgttcgtc acatctcatc tacctcccgg
     1681 ttttaatgaa tacgattttg taccagagtc ctttgatcgt gacaaaacaa ttgcactgat
     1741 aatgaactcc tctggatcta ctgggttacc taagggtgtg gcccttccgc atagaactgc
     1801 ctgcgtcaga ttctcgcatg ccagagatcc tatttttggc aatcaaatca ttccggatac
     1861 tgcgatttta agtgttgttc cattccatca cggttttgga atgtttacta cactcggata
     1921 tttgatatgt ggatttcgag tcgtcttaat gtatagattt gaagaagagc tgtttttacg
     1981 atcccttcag gattacaaaa ttcaaagtgc gttgctagta ccaaccctat tttcattctt
     2041 cgccaaaagc actctgattg acaaatacga tttatctaat ttacacgaaa ttgcttctgg
     2101 gggcgcacct ctttcgaaag aagtcgggga agcggttgca aaacgcttcc atcttccagg
     2161 gatacgacaa ggatatgggc tcactgagac tacatcagct attctgatta cacccgaggg
     2221 ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga aggttgtgga
     2281 tctggatacc gggaaaacgc tgggcgttaa tcagagaggc gaattatgtg tcagaggacc
     2341 tatgattatg tccggttatg taaacaatcc ggaagcgacc aacgccttga ttgacaagga
     2401 tggatggcta cattctggag acatagctta ctgggacgaa gacgaacact tcttcatagt
     2461 tgaccgcttg aagtctttaa ttaaatacaa aggataccag gtggcccccg ctgaattgga
     2521 gtcgatattg ttacaacacc ccaacatctt cgacgcgggc gtggcaggtc ttcccgacga
     2581 tgacgccggt gaacttcccg ccgccgttgt tgttttggag cacggaaaga cgatgacgga
     2641 aaaagagatc gtggattacg tcgccagtca agtaacaacc gcgaaaaagt tgcgcggagg
     2701 agttgtgttt gtggacgaag taccgaaagg tcttaccgga aaactcgacg caagaaaaat
     2761 cagagagatc ctcataaagg ccaagaaggg cggaaagtcc aaattgctcg agggcagagg
     2821 aagtcttcta acatgcggtg acgtggagga gaatcccggc cctgcaccgg gatccatgtc
     2881 cagattagat aaaagtaaag tgattaacag cgcattagag ctgcttaatg aggtcggaat
     2941 cgaaggttta acaacccgta aactcgccca gaagctaggt gtagagcagc ctacattgta
     3001 ttggcacgtg cgcaacaagc agactcttat gaacatgctt tcagaggcaa tactggcgaa
     3061 gcatcacacc cgttcagcac cgttaccgac tgagagttgg cagcagtttc tccaggaaaa
     3121 tgctctgagt ttccgtaaag cattactggt ccatcgtgat ggagcccgat tgcatatagg
     3181 gacctctcct acgccccccc agtttgaaca agcagaggcg caactacgct gtctatgcga
     3241 tgcagggttt tcggtcgagg aggctctttt cattctgcaa tctatcagcc attttacgtt
     3301 gggtgcagta ttagaggagc aagcaacaaa ccagatagaa aataatcatg tgatagacgc
     3361 tgcaccacca ttattacaag aggcatttaa tattcaggcg agaacctctg ctgaaatggc
     3421 cttccatttc gggctgaaat cattaatatt tggattttct gcacagttag atgaaaaaaa
     3481 gcatacaccc attgaggatg gtaataaacc aaaaaagaag agaaagctag cagtgtcagt
     3541 gacatttgaa gatgtggctg tgctctttac tcgggacgag tggaagaagc tggatctgtc
     3601 tcagagaagc ctgtaccgtg aggtgatgct ggagaattac agcaacctgg cctccatggc
     3661 aggattcctg tttaccaaac caaaggtgat ctccctgttg cagcaaggag aggacccgta
     3721 tctcgagggc agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctgc
     3781 accgggatcc atggtgagtg tgattaaacc agaaatgaag atgaggtact acatggacgg
     3841 ctccgtcaat gggcatgagt tcacaattga aggtgaaggc acaggcagac cttacgaggg
     3901 acatcaagag atgacactac gcgtcacaat ggccaagggc gggccaatgc ctttcgcgtt
     3961 tgacttagtg tcacacgtgt tctgttacgg ccacagacct tttactaaat atccagaaga
     4021 gataccagac tatttcaaac aagcatttcc tgaaggcctg tcatgggaaa ggtcgttgga
     4081 gttcgaagat ggtgggtccg cttcagtcag tgcgcatata agccttagag gaaacacctt
     4141 ctaccacaaa tccaaattta ctggggttaa ctttcctgcc gatggtccta tcatgcaaaa
     4201 ccaaagtgtt gattgggagc catcaaccga gaaaattact gccagcgacg gagttctgaa
     4261 gggtgatgtt acgatgtacc taaaacttga aggaggcggc aatcacaaat gccaattcaa
     4321 gactacttac aaggcggcaa aaaagattct taaaatgcca ggaagccatt acatcagcca
     4381 tcgcctcgtc aggaaaaccg aaggcaacat tactgagctg gtagaagatg cagtagctca
     4441 ttcctgatgt acaagtaaag cggccgcaag cttatcgata atcaacctct ggattacaaa
     4501 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     4561 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     4621 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     4681 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     4741 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     4801 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     4861 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     4921 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     4981 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     5041 cggatctccc tttgggccgc ctccccgcct gatcgatacc gtcgactaga gctcgctgat
     5101 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     5161 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     5221 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     5281 gggaggattg ggaagacaat agcaggcatg ctggggagag atctgaggaa cccctagtga
     5341 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc
     5401 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
     5461 gagtggccaa cccccccccc cccccccctg cagcctggcg taatagcgaa gaggcccgca
     5521 ccgatcgccc ttcccaacag ttgcgtagcc tgaatggcga atggcgcgac gcgccctgta
     5581 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5641 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5701 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5761 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     5821 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5881 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     5941 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     6001 acaaaatatt aacgtttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
     6061 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     6121 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     6181 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     6241 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     6301 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     6361 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     6421 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     6481 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     6541 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     6601 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     6661 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     6721 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     6781 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6841 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6901 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6961 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     7021 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     7081 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     7141 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     7201 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     7261 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     7321 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     7381 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     7441 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     7501 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     7561 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     7621 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     7681 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     7741 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7801 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7861 actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc
     7921 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7981 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     8041 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     8101 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     8161 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     8221 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     8281 gcctctcccc gcgcgttggc cgattcatta atgcaggctg cag
//
