| 质粒名称: | pBSDONR P1-P4 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 71744 |
| 存储实验室: | Roger Innes |
|
载体骨架基本信息 |
|
| 载体名称: | pBlueScript SK(+) |
| 载体出品公司: | Stratagene |
| 空载体大小: | 2958 bp |
| 完整质粒大小: | 5371bp |
| 载体类型: | Gateway entry vector for use in multisite Gateway cloning |
| 载体抗性: | Chloramphenicol and Ampicillin |
| 生长温度: | 37 ℃ |
| 宿主菌: | ccdB Survival |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 克隆方法: | 限制性内切酶 |
| 5’测序引物 | T3 |
| 3’测序引物 | T7 |
|
文献引用方法 |
|
| 材料和方法部分: | pBSDONR P1-P4 was a gift from Roger Innes (Addgene plasmid # 71744 ; http://n2t.net/addgene:71744 ; RRID:Addgene_71744) |
| 参考文献部分: | The KEEP ON GOING protein of Arabidopsis recruits the ENHANCED DISEASE RESISTANCE1 protein to trans-Golgi network/early endosome vesicles. Gu Y, Innes RW. Plant Physiol. 2011 Apr;155(4):1827-38. doi: 10.1104/pp.110.171785. Epub 2011 Feb 22. 10.1104/pp.110.171785 PubMed 21343429 |
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载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pBSDONR P1-P4 5371 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION ampicillin resistant derivative of pDONR221 P1-P4 for multisite
GATEWAY cloning.
ACCESSION .
VERSION .
KEYWORDS pBSDONR P1-P4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5371)
AUTHORS Gu Y, Innes RW
TITLE The KEEP ON GOING protein of Arabidopsis recruits the ENHANCED
DISEASE RESISTANCE1 protein to trans-Golgi network/early endosome
vesicles.
JOURNAL Plant Physiol. 2011 Apr;155(4):1827-38. doi: 10.1104/pp.110.171785.
Epub 2011 Feb 22.
PUBMED 21343429
REFERENCE 2 (bases 1 to 5371)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5371
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 7..462
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(94..113)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 304..325
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 588..610
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 602..619
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 603..619
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 629..648
/label=T7
/note="T7 promoter, forward primer"
promoter 629..647
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 669..900
/gene="mutant version of attP"
/label=attP1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction
(pDONR(TM)201 version)"
CDS complement(1296..1601)
/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"
primer_bind complement(1398..1417)
/label=ccdB-fwd
/note="ccdB gene, forward primer"
CDS complement(1951..2604)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG"
primer_bind 2519..2538
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(2605..2707)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
protein_bind complement(2852..3083)
/gene="mutant version of attP"
/label=attP4
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
primer_bind complement(3133..3149)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3133..3149)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(3186..3206)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(3186..3204)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3225..3241)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3225..3241)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3238..3260)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3249..3265
/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(3273..3303)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3318..3339
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3456..3473)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3627..4215)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3707..3726)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4386..5246)
/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"
primer_bind 5009..5028
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5247..5351)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
61 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
121 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
181 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
241 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
301 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
361 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
421 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttccattcgc cattcaggct
481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
601 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac
661 cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga
721 tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gctgaacgag aaacgtaaaa
781 tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta cataatactg
841 taaaacacaa catatccagt cactatgaat caactactta gatggtatta gtgacctgta
901 gtcgaccgac agccttccaa atgttcttcg ggtgatgctg ccaacttagt cgaccgacag
961 ccttccaaat gttcttctca aacggaatcg tcgtatccag cctactcgct attgtcctca
1021 atgccgtatt aaatcataaa aagaaataag aaaaagaggt gcgagcctct tttttgtgtg
1081 acaaaataaa aacatctacc tattcatata cgctagtgtc atagtcctga aaatcatctg
1141 catcaagaac aatttcacaa ctcttatact tttctcttac aagtcgttcg gcttcatctg
1201 gattttcagc ctctatactt actaaacgtg ataaagtttc tgtaatttct actgtatcga
1261 cctgcagact ggctgtgtat aagggagcct gacatttata ttccccagaa catcaggtta
1321 atggcgtttt tgatgtcatt ttcgcggtgg ctgagatcag ccacttcttc cccgataacg
1381 gagaccggca cactggccat atcggtggtc atcatgcgcc agctttcatc cccgatatgc
1441 accaccgggt aaagttcacg ggagacttta tctgacagca gacgtgcact ggccaggggg
1501 atcaccatcc gtcgcccggg cgtgtcaata atatcactct gtacatccac aaacagacga
1561 taacggctct ctcttttata ggtgtaaacc ttaaactgca tttcaccagc ccctgttctc
1621 gtcagcaaaa gagccgttca tttcaataaa ccgggcgacc tcagccatcc cttcctgatt
1681 ttccgctttc cagcgttcgg cacgcagacg acgggcttca ttctgcatgg ttgtgcttac
1741 cagaccggag atattgacat catatatgcc ttgagcaact gatagctgtc gctgtcaact
1801 gtcactgtaa tacgctgctt catagcatac ctctttttga catacttcgg gtatacatat
1861 cagtatatat tcttataccg caaaaatcag cgcgcaaata cgcatactgt tatctggctt
1921 ttagtaagcc ggatccacgc ggcgtttacg ccccccctgc cactcatcgc agtactgttg
1981 taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat gaacctgaat
2041 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg
2101 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
2161 attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
2221 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
2281 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
2341 aacactatcc catatcacca gctcaccgtc tttcattgcc atacggaatt ccggatgagc
2401 attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct tatttttctt
2461 tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg tacattgagc
2521 aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat caacggtggt
2581 atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc tcgataactc
2641 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
2701 ccgatcaacg tctcattttc gccaaaagtt ggcccagggc ttcccggtat caacagggac
2761 accaggattt atttattctg cgaagtgatc ttccgtcaca ggtatttatt cggcgcaaag
2821 tgcgtcgggt gatgctgcca acttagtcga ctacaggtca ctaataccat ctaagtagtt
2881 gattcatagt gactggatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa
2941 aatctaattt aatatattga tatttatatc attttacgtt tctcgttcaa cttttctata
3001 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
3061 cagtcaaaat aaaatcatta tttgccatcc agctgcaggg cggccgcgat atcgaattcc
3121 tgcagcccgg gggatccact agttctagag cggccgccac cgcggtggag ctccagcttt
3181 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
3241 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
3301 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
3361 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
3421 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
3481 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
3541 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
3601 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
3661 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
3721 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
3781 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
3841 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
3901 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
3961 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
4021 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
4081 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
4141 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
4201 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
4261 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
4321 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
4381 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
4441 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
4501 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
4561 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
4621 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
4681 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
4741 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
4801 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
4861 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
4921 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
4981 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
5041 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
5101 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
5161 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
5221 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
5281 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
5341 ggggttccgc gcacatttcc ccgaaaagtg c
//
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