LOCUS       Exported               13426 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  RNAi cloning vector pWALIUM10, complete sequence.
ACCESSION   GU931388
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13426)
  AUTHORS   Ni J-Q., Liu L-P., Perkins LA, Perrimon N.
  TITLE     Vectors for transgenic flies
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 13426)
  AUTHORS   Ni J-Q., Liu L-P., Perkins LA, Perrimon N.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-FEB-2010) Genetics, Harvard Medical School, 77 Avenue 
            Louis Pasteur, Boston, MA 02115, USA
REFERENCE   3  (bases 1 to 13426)
  AUTHORS   Hu Y, Perkins LA.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-JUN-2012) Genetics, Harvard Medical School, 77 Avenue 
            Louis Pasteur, Boston, MA 02115, USA
REFERENCE   4  (bases 1 to 13426)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     On Jun 20, 2012 this sequence version replaced GU931388.1.
FEATURES             Location/Qualifiers
     source          1..13426
                     /organism="RNAi cloning vector pWALIUM10"
                     /lab_host="Drosophila melanogaster"
                     /mol_type="other DNA"
                     /db_xref="taxon:747301"
     rep_origin      complement(6..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     603..619
                     /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_feature    682..4808
                     /label=white
                     /note="white"
     CDS             complement(join(1444..2058,2129..2260,2481..2796,2858..3512,
                     3587..3860,4255..4326))
                     /codon_start=1
                     /product="white"
                     /label=white
                     /note="selectable marker"
                     /protein_id="ADE08350.1"
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLLPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     intron          3513..3586
                     /note="white intron"
     misc_recomb     4851..5218
                     /label=attB docking site
                     /note="attB docking site"
     protein_bind    4923..4992
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     misc_feature    5230..5660
                     /label=gypsy insulator
                     /note="gypsy insulator"
     protein_bind    5230..5660
                     /label=gypsy insulator
                     /bound_moiety="Su(Hw), CP190, and Mod(mdg4)67.2"
                     /note="chromatin insulator from Drosophila"
     misc_recomb     5678..5711
                     /label=LoxP
                     /note="LoxP"
     protein_bind    complement(5678..5711)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    5718..5827
                     /label=5XUAS
                     /note="5XUAS"
     protein_bind    5724..5818
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     misc_recomb     5834..5867
                     /label=LoxP
                     /note="LoxP"
     protein_bind    complement(5834..5867)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    5874..5983
                     /label=5XUAS
                     /note="5XUAS"
     protein_bind    5880..5974
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     regulatory      5990..6249
                     /regulatory_class="promoter"
                     /note="Hsp70 promoter"
     promoter        5998..6236
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     misc_recomb     6277..7988
                     /note="attR; gateway recombination"
     protein_bind    6281..6405
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        6430..6460
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             6514..7173
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             7515..7820
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(7861..7985)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     intron          8008..8154
                     /note="ftz intron"
     misc_recomb     8165..9869
                     /note="attR; gateway recombination"
     protein_bind    8165..8289
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(8330..8635)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(8977..9636)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(9690..9720)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(9745..9869)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     intron          9916..10062
                     /note="ftz intron"
     misc_feature    10086..10785
                     /label=SV40 polyA region
                     /note="SV40 polyA region"
     intron          10162..10227
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             10357..10377
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    10649..10770
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    10792..11222
                     /label=gypsy insulator
                     /note="gypsy insulator"
     protein_bind    10792..11222
                     /label=gypsy insulator
                     /bound_moiety="Su(Hw), CP190, and Mod(mdg4)67.2"
                     /note="chromatin insulator from Drosophila"
     promoter        complement(11241..11259)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11280..11296)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    11304..11320
                     /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(11328..11358)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11373..11394
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(11682..12270)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(12441..13301)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    12444..13103
                     /label=AmpR
                     /note="AmpR"
     promoter        complement(13302..13406)
                     /gene="bla"
                     /label=AmpR promoter