Basic Vector Information
- Vector Name:
- pP-ccdB
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 6130 bp
- Type:
- MISSA donor vector
- Replication origin:
- ori
- Source/Author:
- Chen QJ, Xie M, Ma XX, Dong L, Chen J, Wang XC.
- Promoter:
- sacB
pP-ccdB 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 ℃.
pP-ccdB vector Sequence
LOCUS 40924_33967 6130 bp DNA circular SYN 18-DEC-2018 DEFINITION MISSA donor vector pP-ccdB, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6130) AUTHORS Chen QJ, Xie M, Ma XX, Dong L, Chen J, Wang XC. TITLE MISSA is a highly efficient in vivo DNA assembly method for plant multiple-gene transformation JOURNAL Plant Physiol. 153 (1), 41-51 (2010) PUBMED 20200068 REFERENCE 2 (bases 1 to 6130) AUTHORS Chen Q-J., Xie M, Ma X-X., Dong L, Chen J, Wang X-C. TITLE Direct Submission JOURNAL Submitted (27-JAN-2010) State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, No. 2 West Yuanmingyuan Road, Haidian, Beijing 100193, China REFERENCE 3 (bases 1 to 6130) TITLE Direct Submission REFERENCE 4 (bases 1 to 6130) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant Physiol."; date: "2010"; volume: "153"; issue: "1"; pages: "41-51" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (27-JAN-2010) State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, No. 2 West Yuanmingyuan Road, Haidian, Beijing 100193, China" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..6130 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind complement(1..34) /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." terminator 100..186 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 278..305 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" primer_bind 373..389 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 397..415 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" protein_bind 424..655 /label=attP1 /note="recombination site for the Gateway(R) BP reaction" terminator complement(733..760) /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator complement(852..938) /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" rep_origin complement(1103..1691) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(1844..2146) /codon_start=1 /label=ccdB /note="CcdB, a bacterial toxin that poisons DNA gyrase" /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRIVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" protein_bind complement(2305..2536) /label=attP2 /note="recombination site for the Gateway(R) BP reaction (pDONR(TM)221 version)" promoter complement(2554..2572) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2584..2600) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" CDS complement(2674..4092) /codon_start=1 /label=SacB /note="secreted levansucrase that renders bacterial growth sensitive to sucrose" /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV VKDSILEQGQLTVNK" promoter complement(4093..4538) /label=sacB promoter /note="sacB promoter and control region" CDS complement(4676..5332) /codon_start=1 /label=CmR /note="chloramphenicol acetyltransferase" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" promoter complement(5385..5415) /label=lac UV5 promoter /note="E. coli lac promoter with an 'up' mutation" rep_origin 5442..5795 /label=oriR6Kgamma /note="oriR6Kgamma" oriT 5937..6046 /label=oriT /note="incP origin of transfer"
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