LOCUS       Exported                4828 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  SiRNA vector pSUPER-hSyn-EGFP-CytB-AS, complete sequence.
ACCESSION   AY640629
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4828)
  AUTHORS   Michel U, Malik I, Ebert S, Bahr M, Kugler S.
  TITLE     Long-term in vivo and in vitro AAV-2-mediated RNA interference in 
            rat retinal ganglion cells and cultured primary neurons
  JOURNAL   Biochem. Biophys. Res. Commun. 326 (2), 307-312 (2005)
  PUBMED    15582578
REFERENCE   2  (bases 1 to 4828)
  AUTHORS   Michel U, Kuegler S.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-MAY-2004) Neurology, University of Goettingen, Waldweg
            33, Goettingen 37073, Germany
REFERENCE   3  (bases 1 to 4828)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4828
                     /organism="siRNA vector pSUPER-hSyn-EGFP-CytB-AS"
                     /mol_type="other DNA"
                     /db_xref="taxon:297631"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      135..441
                     /label=F1-Ori
                     /note="F1-Ori"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      625..645
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     677..693
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(694..854)
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40-pA; SV40 polyadenylation sequence"
     polyA_signal    707..841
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     intron          complement(854..998)
                     /note="SV40 chimeric intron derived from pCI-NEO"
     intron          857..990
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             complement(1002..1721)
                     /codon_start=1
                     /gene="EGFP"
                     /product="enhanced green fluorescent protein"
                     /label=EGFP
                     /note="derived from Clontechs pEGFP-N1"
                     /protein_id="AAV73960.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     gene            complement(1002..1721)
                     /gene="EGFP"
                     /label=EGFP
     regulatory      1718..1727
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    complement(1740..1758)
                     /note="MCS; multiple cloning site"
     regulatory      complement(1758..2227)
                     /regulatory_class="promoter"
                     /note="HSYN-promoter; human synapsin 1 gene promoter (-422 
                     to +53)"
     promoter        complement(1764..2211)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      2239..2454
                     /regulatory_class="promoter"
                     /note="disabled human H1 RNA promoter"
     promoter        2239..2453
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     misc_feature    2455..2586
                     /note="Cytb-AS; partial human cytochrome B antisense 
                     sequence"
     primer_bind     complement(2594..2610)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(2639..2657)
                     /regulatory_class="promoter"
                     /note="T3 promoter"
     promoter        complement(2640..2658)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2679..2695)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2703..2719
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(2727..2757)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2772..2793
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      3025..3692
                     /label=pUC Ori
                     /note="pUC Ori"
     rep_origin      complement(3081..3669)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3840..4700)
                     /codon_start=1
                     /gene="Amp"
                     /product="beta-lactamase"
                     /label=Amp
                     /note="ampicillin resistance"
                     /protein_id="AAV73961.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3840..4700)
                     /gene="Amp"
                     /label=Amp
     promoter        complement(4701..4805)
                     /gene="bla"
                     /label=AmpR promoter