LOCUS       Exported                8687 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pAAV-TRE-CAG-Fluc, complete sequence.
ACCESSION   KC152482
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8687
                     /organism="Expression vector pAAV-TRE-CAG-Fluc"
                     /mol_type="other DNA"
                     /db_xref="taxon:1337869"
     repeat_region   19..162
                     /label=AAV2 ITR
                     /note="AAV2 ITR"
     misc_feature    245..542
                     /label=tetracycline responsive element: TRE
                     /note="tetracycline responsive element: TRE"
     protein_bind    258..528
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     regulatory      543..559
                     /regulatory_class="promoter"
                     /note="CMVmin"
     regulatory      564..1504
                     /regulatory_class="promoter"
                     /note="CMV earlyenhancer/chicken beta actin promoter: CAG"
     promoter        814..1091
                     /label=chicken beta-actin promoter
     misc_feature    1521..3173
                     /label=firefly luciferase
                     /note="firefly luciferase"
     CDS             1521..3170
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     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"
     misc_feature    3240..4088
                     /label=tTSKid
                     /note="tTSKid"
     misc_feature    3240..3872
                     /label=Tet repressor: tetR[B/E]
                     /note="Tet repressor: tetR[B/E]"
     misc_feature    3873..3890
                     /label=nuclear localization signal: NLS
                     /note="nuclear localization signal: NLS"
     misc_feature    3897..4082
                     /note="KRAB (Kruppel-associated box) domain; from Rat zinc 
                     finger protein (kid-1)"
     CDS             3897..4022
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the rat zinc finger protein Kid-1 
                     (Witzgall et al., 1994)"
                     /label=KRAB-A
                     /translation="VSVTFEDVAVLFTRDEWKKLDLSQRSLYREVMLENYSNLASM"
     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..4811
                     /codon_start=1
                     /gene="mKO"
                     /product="monomeric Kusabira-orange"
                     /label=mKO
                     /note="fluorescent protein"
                     /protein_id="AGM53875.1"
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAKGGPMPFAFDLVSHVFCYGHRPFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKKILKMPGSHYISHRLVRKTEGNITELVEDAVAHS"
     gene            4155..4811
                     /gene="mKO"
                     /label=mKO
     ncRNA           4842..5438
                     /product="post-transcriptional regulatory element: WPRE"
                     /label=other
                     /ncRNA_class="other"
     misc_feature    4844..5432
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(5315..5326)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    5471..5681
                     /label=bovine growth hormone polyA signal: bGH pA
                     /note="bovine growth hormone polyA signal: bGH pA"
     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
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6720..6824
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6825..7685
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             7023..7685
                     /codon_start=1
                     /gene="Bla"
                     /product="beta-lactamase"
                     /label=Bla
                     /note="ampicillin resistance"
                     /protein_id="AGM53876.1"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     gene            7023..7685
                     /gene="Bla"
                     /label=Bla
     rep_origin      7856..8444
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"