pAAV-CAG-Fluc vector (V009683)

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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"