LOCUS       Exported                5357 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  CRIMIC donor vector pM14, complete sequence.
ACCESSION   MH753008
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5357)
  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 5357)
  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 5357)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5357
                     /organism="CRIMIC donor vector pM14"
                     /mol_type="other DNA"
                     /db_xref="taxon:2340942"
     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     710..930
                     /label=attP phage phi-C31 attachment site
                     /note="attP phage phi-C31 attachment site"
     protein_bind    775..874
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     misc_recomb     931..964
                     /label=FRT (minimal)
                     /note="FRT (minimal)"
     protein_bind    931..964
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    971..1070
                     /label=MHC splice acceptor site
                     /note="MHC splice acceptor site"
     regulatory      1071..1088
                     /regulatory_class="terminator"
                     /note="stop codons in all reading frames"
     polyA_signal    1089..1223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      1093..1284
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 polyadenylation signal"
     regulatory      1331..1596
                     /regulatory_class="promoter"
                     /note="3XP3 promoter"
     regulatory      1597..1839
                     /regulatory_class="promoter"
                     /note="Hsp70 promoter"
     promoter        1602..1839
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     regulatory      1846..1855
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1852..2571
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=enhanced GFP
                     /protein_id="AYC12532.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    2572..2706
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      2576..2767
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 polyadenylation signal"
     misc_recomb     2814..2847
                     /label=FRT (minimal)
                     /note="FRT (minimal)"
     protein_bind    2814..2847
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_recomb     complement(2848..3068)
                     /label=attP phage phi-C31 attachment site
                     /note="attP phage phi-C31 attachment site"
     protein_bind    2904..3003
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     primer_bind     complement(3123..3139)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3169..3187)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3208..3224)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3232..3248
                     /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(3256..3286)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3301..3322
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3610..4198)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4369..5229)
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="AYC12533.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5230..5334)
                     /gene="bla"
                     /label=AmpR promoter