Lee PT, Zirin J, Kanca O, Lin WW, Schulze KL, Li-Kroeger D, Tao R, Devereaux C, Hu Y, Chung V, Fang Y, He Y, Pan H, Ge M, Zuo Z, Housden BE, Mohr SE, Yamamoto S, Levis RW, Spradling AC, Perrimon N, Be
pM36 载体载体图谱
pM36 载体载体序列
LOCUS Exported 5337 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION CRIMIC donor vector pM36, complete sequence.
ACCESSION MH753009
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5337)
AUTHORS Lee PT, Zirin J, Kanca O, Lin WW, Schulze KL, Li-Kroeger D, Tao R,
Devereaux C, Hu Y, Chung V, Fang Y, He Y, Pan H, Ge M, Zuo Z,
Housden BE, Mohr SE, Yamamoto S, Levis RW, Spradling AC, Perrimon N,
Bellen HJ.
TITLE A gene-specific T2A-GAL4 library for Drosophila
JOURNAL Elife 7, e35574 (2018)
PUBMED 29565247
REFERENCE 2 (bases 1 to 5337)
AUTHORS Lee P-T., Zirin J, Kanca O, Lin W-W., Schulze KL, Li-Kroeger D, Tao
R, Devereaux C, Hu Y, Chung V, Fang Y, He Y, Pan H, Ge M, Zuo Z,
Housden BE, Mohr SE, Yamamoto S, Levis RW, Spradling AC, Perrimon N,
Bellen HJ.
TITLE Direct Submission
JOURNAL Submitted (16-AUG-2018) HHMI, Molecular & Human Genetics, Baylor
College of Medicine, 1125 Moursund St. Ste. 1125, Houston, TX 77030,
USA
REFERENCE 3 (bases 1 to 5337)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..5337
/organism="CRIMIC donor vector pM36"
/mol_type="other DNA"
/db_xref="taxon:2340943"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 600..616
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 626..644
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_recomb 733..953
/label=attP phage phi-C31 attachment site
/note="attP phage phi-C31 attachment site"
protein_bind 798..897
/label=phage phi-C31 attP
/bound_moiety="phage phi-C31 integrase"
/note="attachment site of phage phi-C31"
misc_feature 960..1059
/label=MHC splice acceptor site
/note="MHC splice acceptor site"
regulatory 1060..1077
/regulatory_class="terminator"
/note="stop codons in all reading frames"
polyA_signal 1078..1212
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
regulatory 1082..1273
/regulatory_class="polyA_signal_sequence"
/note="SV40 polyadenylation signal"
regulatory 1320..1585
/regulatory_class="promoter"
/note="3XP3 promoter"
regulatory 1586..1828
/regulatory_class="promoter"
/note="Hsp70 promoter"
promoter 1591..1828
/label=hsp70 promoter
/note="Drosophila melanogaster hsp70Bb promoter"
regulatory 1835..1844
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1841..2560
/codon_start=1
/product="enhanced GFP"
/label=enhanced GFP
/protein_id="AYC12534.1"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
polyA_signal 2561..2695
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
regulatory 2565..2756
/regulatory_class="polyA_signal_sequence"
/note="SV40 polyadenylation signal"
misc_recomb complement(2803..3023)
/label=attP phage phi-C31 attachment site
/note="attP phage phi-C31 attachment site"
protein_bind 2859..2958
/label=phage phi-C31 attP
/bound_moiety="phage phi-C31 integrase"
/note="attachment site of phage phi-C31"
primer_bind complement(3103..3119)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(3149..3167)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3188..3204)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3212..3228
/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(3236..3266)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3281..3302
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3590..4178)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4349..5209)
/codon_start=1
/product="AmpR"
/label=AmpR
/protein_id="AYC12535.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5210..5314)
/gene="bla"
/label=AmpR promoter
Lee PT, Zirin J, Kanca O, Lin WW, Schulze KL, Li-Kroeger D, Tao R, Devereaux C, Hu Y, Chung V, Fang Y, He Y, Pan H, Ge M, Zuo Z, Housden BE, Mohr SE, Yamamoto S, Levis RW, Spradling AC, Perrimon N, Be
pM36 载体载体图谱
pM36 载体载体序列
LOCUS Exported 5337 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION CRIMIC donor vector pM36, complete sequence.
ACCESSION MH753009
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5337)
AUTHORS Lee PT, Zirin J, Kanca O, Lin WW, Schulze KL, Li-Kroeger D, Tao R,
Devereaux C, Hu Y, Chung V, Fang Y, He Y, Pan H, Ge M, Zuo Z,
Housden BE, Mohr SE, Yamamoto S, Levis RW, Spradling AC, Perrimon N,
Bellen HJ.
TITLE A gene-specific T2A-GAL4 library for Drosophila
JOURNAL Elife 7, e35574 (2018)
PUBMED 29565247
REFERENCE 2 (bases 1 to 5337)
AUTHORS Lee P-T., Zirin J, Kanca O, Lin W-W., Schulze KL, Li-Kroeger D, Tao
R, Devereaux C, Hu Y, Chung V, Fang Y, He Y, Pan H, Ge M, Zuo Z,
Housden BE, Mohr SE, Yamamoto S, Levis RW, Spradling AC, Perrimon N,
Bellen HJ.
TITLE Direct Submission
JOURNAL Submitted (16-AUG-2018) HHMI, Molecular & Human Genetics, Baylor
College of Medicine, 1125 Moursund St. Ste. 1125, Houston, TX 77030,
USA
REFERENCE 3 (bases 1 to 5337)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..5337
/organism="CRIMIC donor vector pM36"
/mol_type="other DNA"
/db_xref="taxon:2340943"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 600..616
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 626..644
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_recomb 733..953
/label=attP phage phi-C31 attachment site
/note="attP phage phi-C31 attachment site"
protein_bind 798..897
/label=phage phi-C31 attP
/bound_moiety="phage phi-C31 integrase"
/note="attachment site of phage phi-C31"
misc_feature 960..1059
/label=MHC splice acceptor site
/note="MHC splice acceptor site"
regulatory 1060..1077
/regulatory_class="terminator"
/note="stop codons in all reading frames"
polyA_signal 1078..1212
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
regulatory 1082..1273
/regulatory_class="polyA_signal_sequence"
/note="SV40 polyadenylation signal"
regulatory 1320..1585
/regulatory_class="promoter"
/note="3XP3 promoter"
regulatory 1586..1828
/regulatory_class="promoter"
/note="Hsp70 promoter"
promoter 1591..1828
/label=hsp70 promoter
/note="Drosophila melanogaster hsp70Bb promoter"
regulatory 1835..1844
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1841..2560
/codon_start=1
/product="enhanced GFP"
/label=enhanced GFP
/protein_id="AYC12534.1"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
polyA_signal 2561..2695
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
regulatory 2565..2756
/regulatory_class="polyA_signal_sequence"
/note="SV40 polyadenylation signal"
misc_recomb complement(2803..3023)
/label=attP phage phi-C31 attachment site
/note="attP phage phi-C31 attachment site"
protein_bind 2859..2958
/label=phage phi-C31 attP
/bound_moiety="phage phi-C31 integrase"
/note="attachment site of phage phi-C31"
primer_bind complement(3103..3119)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(3149..3167)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3188..3204)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3212..3228
/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(3236..3266)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3281..3302
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3590..4178)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4349..5209)
/codon_start=1
/product="AmpR"
/label=AmpR
/protein_id="AYC12535.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5210..5314)
/gene="bla"
/label=AmpR promoter