LOCUS       Exported                7830 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  CRIMIC donor vector pM37(2), complete sequence.
ACCESSION   MH753012
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7830)
  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 7830)
  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 7830)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7830
                     /organism="CRIMIC donor vector pM37(2)"
                     /mol_type="other DNA"
                     /db_xref="taxon:2340946"
     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     775..874
                     /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"
     misc_feature    1117..1170
                     /label=similar to T2A peptide
                     /note="similar to T2A peptide"
     CDS             1117..1170
                     /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             1171..3816
                     /codon_start=1
                     /product="GAL4"
                     /label=GAL4
                     /protein_id="AYC12542.1"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVSIDSAAHHDNSTIPLDF
                     MPRDALHGFDWSEEDDMSDGLPFLKTDPNNNGFFGDGSLLCILRSIGFKPENYTNSNVN
                     RLPTMITDRYTLASRSTTSRLLQSYLNNFHPYCPIVHSPTLMMLYNNQIEIASKDQWQI
                     LFNCILAIGAWCIEGESTDIDVFYYQNAKSHLTSKVFESGSIILVTALHLLSRYTQWRQ
                     KTNTSYNFHSFSIRMAISLGLNRDLPSSFSDSSILEQRRRIWWSVYSWEIQLSLLYGRS
                     IQLSQNTISFPSSVDDVQRTTTGPTIYHGIIETARLLQVFTKIYELDKTVTAEKSPICA
                     KKCLMICNEIEEVSRQAPKFLQMDISTTALTNLLKEHPWLSFTRFELKWKQLSLIIYVL
                     RDFFTNFTQKKSQLEQDQNDHQSYEVKRCSIMLSDAAQRTVMSVSSYMDNHNVTPYFAW
                     NCSYYLFNAVLVPIKTLLSNSKSNAENNETAQLLQQINTVLMLLKKLATFKIQTCEKYI
                     QVLEEVCAPFLLSQCAIPLPHISYNNSNGSAIKNIVGSATIAQYPTLPEENVNNISVKY
                     VSPGSVGPSPVPLKSGASFSDLVKLLSNRPPSRNSPVTIPRSTPSHRSVTPFLGQQQQL
                     QSLVPLTPSALFGGANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     regulatory      3832..3966
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 polyadenylation signal"
     polyA_signal    3832..3966
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      4092..4297
                     /regulatory_class="promoter"
                     /note="3XP3 promoter"
     regulatory      4319..4328
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4325..5044
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=enhanced GFP
                     /protein_id="AYC12543.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      5045..5179
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 polyadenylation signal"
     polyA_signal    5045..5179
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_recomb     5287..5320
                     /label=FRT (minimal)
                     /note="FRT (minimal)"
     protein_bind    5287..5320
                     /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(5377..5476)
                     /label=attP phage phi-C31 attachment site
                     /note="attP phage phi-C31 attachment site"
     protein_bind    5377..5476
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     primer_bind     complement(5596..5612)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5642..5660)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5681..5697)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5705..5721
                     /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(5729..5759)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5774..5795
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6083..6671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6842..7702)
                     /codon_start=1
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="AYC12544.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7703..7807)
                     /gene="bla"
                     /label=AmpR promoter