pWALIUM10 vector (V002214)

Basic Vector Information

Vector Name:
pWALIUM10
Antibiotic Resistance:
Ampicillin
Length:
13426 bp
Type:
RNAi cloning vector
Replication origin:
ori
Source/Author:
Ni J-Q., Liu L-P., Perkins LA, Perrimon N.
Promoter:
hsp70

pWALIUM10 vector Map

pWALIUM1013426 bp600120018002400300036004200480054006000660072007800840090009600102001080011400120001260013200f1 oriM13 fwdT7 promoterwhiteattBgypsy insulatorloxP5X UASLoxP5XUAShsp70 promoterattR1lac UV5 promoterCmRccdBattR2ftz intronattR2ccdBCmRlac UV5 promoterattR1ftz intronsmall t intronSV40 NLSSV40 poly(A) signalgypsy insulatorT3 promoterM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoter

pWALIUM10 vector Sequence

LOCUS       40924_46168       13426 bp DNA     circular SYN 18-DEC-2018
DEFINITION  RNAi cloning vector pWALIUM10, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 13426)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-FEB-2010) Genetics, Harvard Medical School, 77 Avenue Louis 
            Pasteur, Boston, MA 02115, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (20-JUN-2012) Genetics, Harvard Medical School, 77 Avenue Louis 
            Pasteur, Boston, MA 02115, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     On Jun 20, 2012 this sequence version replaced GU931388.1.
FEATURES             Location/Qualifiers
     source          1..13426
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     protein_bind    4923..4992
                     /label=attB
                     /note="attB site for the phi-C31 integrase (Groth et al.,
                     2000)"
     protein_bind    5230..5660
                     /label=gypsy insulator
                     /note="chromatin insulator from Drosophila"
     protein_bind    5678..5711
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5724..5818
                     /label=5X UAS
                     /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)"
     promoter        5998..6236
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     protein_bind    6281..6405
                     /label=attR1
                     /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..7170
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             7515..7817
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(7861..7985)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     intron          8008..8154
                     /note="ftz intron"
     protein_bind    8165..8289
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(8333..8635)
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             complement(8980..9636)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(9690..9720)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    complement(9745..9869)
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     intron          9916..10062
                     /note="ftz intron"
     intron          10162..10227
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             10357..10377
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    10649..10783
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    10792..11222
                     /label=gypsy insulator
                     /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    complement(11373..11394)
                     /label=CAP binding site
                     /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(12444..13301)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(13302..13406)
                     /label=AmpR promoter

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