LOCUS       Exported               12502 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  RNAi cloning vector pRISE DNA, complete sequence.
ACCESSION   AB262394
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12502)
  AUTHORS   Kondo T, Inagaki S, Yasuda K, Kageyama Y.
  TITLE     Rapid construction of Drosophila RNAi transgenes using pRISE, a 
            P-element-mediated transformation vector exploiting an in vitro 
            recombination system
  JOURNAL   Genes Genet. Syst. 81 (2), 129-134 (2006)
  PUBMED    16755136
REFERENCE   2  (bases 1 to 12502)
  AUTHORS   Kondo T, Inagaki S, Kageyama Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JUN-2006) Takefumi Kondo, Nara Inst. Sci. Tech., Grad.
            Sch. Biol. Sci.; 8916-5 Takayama, Ikoma, Nara 630-0192, Japan 
            (E-mail:ta-kondo@bs.naist.jp, Tel:81-743-72-5554, 
            Fax:81-743-72-5559)
REFERENCE   3  (bases 1 to 12502)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12502
                     /organism="RNAi cloning vector pRISE"
                     /mol_type="other DNA"
                     /db_xref="taxon:389357"
     misc_feature    1..5459
                     /note="Corresponging to 1..5608 of pUAST backbone 
                     (http://flybase.bio.indiana.edu/.bin/ 
                     tpseq.html?FBmc0000383)(2006/06/08). There are some 
                     spontaneous mutations compared to the original pUAST 
                     sequence: A 154 bp region at the position 5026-5180 of the 
                     original pUAST is deleted. 4645G of the original pUAST has 
                     been substituted by A, which result in an additional EcoRV 
                     site. There may be other mutations but not extensively 
                     checked."
     repeat_region   1..587
                     /label=P5
                     /note="P5"
     misc_feature    1..586
                     /label=P element 5 end
     misc_feature    complement(602..4728)
                     /label=mini-white
                     /note="mini-white"
     CDS             complement(join(1364..1978,2049..2180,2401..2716,2778..3432,
                     3507..3780,4175..4246))
                     /codon_start=1
                     /gene="white"
                     /product="Drosophila white gene eye color pigment"
                     /label=mini-white
                     /note="This is a modified version of the white gene lacking
                     part of the first intron."
                     /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY
                     GTLLPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH
                     IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG
                     QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL
                     SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ
                     VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP
                     TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP
                     ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG
                     VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL
                     FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV
                     GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS
                     NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE"
     regulatory      complement(4756..5452)
                     /regulatory_class="terminator"
                     /note="SV40 terminator"
     polyA_signal    4756..4890
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5161..5181)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          5311..5376
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     misc_feature    5460..7164
                     /note="Gateway Reading Frame Cassette A 2: attR1 ccdB CmR 
                     attR2"
     protein_bind    5460..5584
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        5609..5639
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             5693..6352
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             6694..6999
                     /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(7040..7164)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    complement(7175..7321)
                     /label=fushi-tarazu intron
                     /note="fushi-tarazu intron"
     misc_feature    complement(7341..9052)
                     /note="Gateway Reading Frame cassette A 1 : attR1 ccdB CmR 
                     attR2"
     protein_bind    7344..7468
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(7509..7814)
                     /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(8156..8815)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(8869..8899)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(8924..9048)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    9053..12502
                     /note="Corresponding to 5617..9053 of pUAST backbone 
                     (http://flybase.bio.indiana.edu/.bin/ 
                     tpseq.html?FBmc0000383)(2006/06/08). There are some 
                     spontaneous mutations compared to the original pUAST 
                     sequence: A 14 bp fragment CTCCCAACCGTTGG (9482..9495 of 
                     pRISE) is inserted into the original pUAST at the potision 
                     6045. There may be other mutations but not extensively 
                     checked."
     promoter        complement(9098..9336)
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     regulatory      complement(9099..9341)
                     /regulatory_class="promoter"
                     /note="Hsp70Bb promoter"
     misc_feature    complement(9342..9463)
                     /label=5xUAS
                     /note="5xUAS"
     protein_bind    9355..9449
                     /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)"
     repeat_region   9510..9742
                     /label=P3
                     /note="P3"
     misc_feature    9510..9742
                     /label=P element 3 end
                     /note="P element 3 end"
     misc_feature    10245..12274
                     /label=pUC8
                     /note="pUC8"
     promoter        10544..10648
                     /gene="bla"
                     /label=AmpR promoter
     CDS             10649..11509
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      11680..12268
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"