Basic Vector Information
piggyBac_RB vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
piggyBac_RB vector Sequence
LOCUS 40924_25426 10035 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector piggyBac_RB, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10035) AUTHORS Thibault ST, Singer MA, Miyazaki WY, Milash B, Dompe NA, Singh CM, Buchholz R, Demsky M, Fawcett R, Francis-Lang HL, Ryner L, Cheung LM, Chong A, Erickson C, Fisher WW, Greer K, Hartouni SR, Howie E, Jakkula L, Joo D, Killpack K, Laufer A, Mazzotta J, Smith RD, Stevens LM, Stuber C, Tan LR, Ventura R, Woo A, Zakrajsek I, Zhao L, Chen F, Swimmer C, Kopczynski C, Duyk G, Winberg ML, Margolis J. TITLE A complementary transposon tool kit for Drosophila melanogaster using P and piggyBac JOURNAL Nat. Genet. 36 (3), 283-287 (2004) PUBMED 14981521 REFERENCE 2 (bases 1 to 10035) AUTHORS Dompe NA, Buchholz R, Ryner L, Miyazaki WY, Kopczynski C, Thibault ST. TITLE Direct Submission JOURNAL Submitted (19-DEC-2003) Exelixis, Inc., 170 Harbor Way, PO Box 511, South San Francisco, CA 94083, USA REFERENCE 3 (bases 1 to 10035) TITLE Direct Submission REFERENCE 4 (bases 1 to 10035) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Genet."; date: "2004"; volume: "36"; issue: "3"; pages: "283-287" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (19-DEC-2003) Exelixis, Inc., 170 Harbor Way, PO Box 511, South San Francisco, CA 94083, USA" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..10035 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 96..200 /label=AmpR promoter CDS 201..1058 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 1232..1820 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 2108..2129 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 2144..2174 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2182..2198 /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)." primer_bind 2206..2222 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" gene 3100..7236 /label=mini-white /note="This modified version of the white gene lacks part of the first intron." protein_bind 7430..7477 /label=FRT /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." primer_bind complement(9641..9657) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants"
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