LOCUS Exported 4535 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Rabies virus-derived expression vector pTIT-N, complete sequence.
ACCESSION GU299212
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4535)
AUTHORS Finke S, Conzelmann KK.
TITLE Virus promoters determine interference by defective RNAs: selective
amplification of mini-RNA vectors and rescue from cDNA by a 3
copy-back ambisense rabies virus
JOURNAL J. Virol. 73 (5), 3818-3825 (1999)
PUBMED 10196276
REFERENCE 2 (bases 1 to 4535)
AUTHORS Wickersham IR, Sullivan HA, Seung HS.
TITLE Production of glycoprotein-deleted rabies viruses for monosynaptic
tracing and high-level gene expression in neurons
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 4535)
AUTHORS Wickersham IR.
TITLE Direct Submission
JOURNAL Submitted (06-DEC-2009) Brain and Cognitive Sciences, Howard Hughes
Medical Institute and Massachusetts Institute of Technology, 43
Vassar St., Cambridge, MA 02139, USA
REFERENCE 4 (bases 1 to 4535)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..4535
/organism="Rabies virus-derived expression vector pTIT-N"
/mol_type="other DNA"
/db_xref="taxon:715317"
primer_bind 148..164
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
regulatory 171..190
/regulatory_class="promoter"
/note="T7"
promoter 171..189
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 204..703
/label=EMCV IRES
/note="EMCV IRES"
misc_feature 240..701
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 706..2058
/codon_start=1
/gene="N"
/product="SAD B19 nucleocapsid protein"
/label=N
/protein_id="ADB43104.1"
/translation="MDADKIVFKVNNQVVSLKPEIIVDQYEYKYPAIKDLKKPCITLGK
APDLNKAYKSVLSGMSAAKLNPDDVCSYLAAAMQFFEGTCPEDWTSYGIVIARKGDKIT
PGSLVEIKRTDVEGNWALTGGMELTRDPTVPEHASLVGLLLSLYRLSKISGQNTGNYKT
NIADRIEQIFETAPFVKIVEHHTLMTTHKMCANWSTIPNFRFLAGTYDMFFSRIEHLYS
AIRVGTVVTAYEDCSGLVSFTGFIKQINLTAREAILYFFHKNFEEEIRRMFEPGQETAV
PHSYFIHFRSLGLSGKSPYSSNAVGHVFNLIHFVGCYMGQVRSLNATVIAACAPHEMSV
LGGYLGEEFFGKGTFERRFFRDEKELQEYEAAELTKTDVALADDGTVNSDDEDYFSGET
RSPEAVYTRIMMNGGRLKRSHIRRYVSVSSNHQARPNSFAEFLNKTYSSDS"
gene 706..2058
/gene="N"
/label=N
terminator 2221..2268
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind complement(2291..2307)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(2344..2362)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2383..2399)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2407..2423
/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(2431..2461)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2476..2497
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(2785..3373)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3544..4404)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=bla
/note="ampicillin-resistance protein"
/protein_id="ADB43105.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene complement(3544..4404)
/gene="bla"
/label=bla
promoter complement(4405..4509)
/gene="bla"
/label=AmpR promoter
LOCUS Exported 4535 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Rabies virus-derived expression vector pTIT-N, complete sequence.
ACCESSION GU299212
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4535)
AUTHORS Finke S, Conzelmann KK.
TITLE Virus promoters determine interference by defective RNAs: selective
amplification of mini-RNA vectors and rescue from cDNA by a 3
copy-back ambisense rabies virus
JOURNAL J. Virol. 73 (5), 3818-3825 (1999)
PUBMED 10196276
REFERENCE 2 (bases 1 to 4535)
AUTHORS Wickersham IR, Sullivan HA, Seung HS.
TITLE Production of glycoprotein-deleted rabies viruses for monosynaptic
tracing and high-level gene expression in neurons
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 4535)
AUTHORS Wickersham IR.
TITLE Direct Submission
JOURNAL Submitted (06-DEC-2009) Brain and Cognitive Sciences, Howard Hughes
Medical Institute and Massachusetts Institute of Technology, 43
Vassar St., Cambridge, MA 02139, USA
REFERENCE 4 (bases 1 to 4535)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..4535
/organism="Rabies virus-derived expression vector pTIT-N"
/mol_type="other DNA"
/db_xref="taxon:715317"
primer_bind 148..164
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
regulatory 171..190
/regulatory_class="promoter"
/note="T7"
promoter 171..189
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 204..703
/label=EMCV IRES
/note="EMCV IRES"
misc_feature 240..701
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 706..2058
/codon_start=1
/gene="N"
/product="SAD B19 nucleocapsid protein"
/label=N
/protein_id="ADB43104.1"
/translation="MDADKIVFKVNNQVVSLKPEIIVDQYEYKYPAIKDLKKPCITLGK
APDLNKAYKSVLSGMSAAKLNPDDVCSYLAAAMQFFEGTCPEDWTSYGIVIARKGDKIT
PGSLVEIKRTDVEGNWALTGGMELTRDPTVPEHASLVGLLLSLYRLSKISGQNTGNYKT
NIADRIEQIFETAPFVKIVEHHTLMTTHKMCANWSTIPNFRFLAGTYDMFFSRIEHLYS
AIRVGTVVTAYEDCSGLVSFTGFIKQINLTAREAILYFFHKNFEEEIRRMFEPGQETAV
PHSYFIHFRSLGLSGKSPYSSNAVGHVFNLIHFVGCYMGQVRSLNATVIAACAPHEMSV
LGGYLGEEFFGKGTFERRFFRDEKELQEYEAAELTKTDVALADDGTVNSDDEDYFSGET
RSPEAVYTRIMMNGGRLKRSHIRRYVSVSSNHQARPNSFAEFLNKTYSSDS"
gene 706..2058
/gene="N"
/label=N
terminator 2221..2268
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind complement(2291..2307)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(2344..2362)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2383..2399)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2407..2423
/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(2431..2461)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2476..2497
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(2785..3373)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3544..4404)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=bla
/note="ampicillin-resistance protein"
/protein_id="ADB43105.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
gene complement(3544..4404)
/gene="bla"
/label=bla
promoter complement(4405..4509)
/gene="bla"
/label=AmpR promoter