| 质粒名称: | pSK+KanaRpsL |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 20871 |
| 存储实验室: | Jiyue Zhu |
|
载体骨架基本信息 |
|
| 载体名称: | pBluescript SK+ |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素和卡那霉素 |
| 空载体大小: | 2935 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | KanamycinResistance-RpsL,KanaRpsL,streptomycin |
| 物种来源: | E.coli |
| 插入大小: | 1743 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Hind III(未破坏) |
| 3' 克隆位点: | BamH I(未破坏) |
| 5' 测序引物 | T7 |
| 3' 测序引物 | T3 |
|
文献引用方法 |
|
| 材料和方法部分: | pSK+KanaRpsL was a gift from Jiyue Zhu (Addgene plasmid # 20871 ; http://n2t.net/addgene:20871 ; RRID:Addgene_20871) |
| 参考文献部分: | A New Positive/Negative Selection Scheme for Precise BAC Recombineering. Wang S, Zhao Y, Leiby M, Zhu J. Mol Biotechnol. 2009 Jan 22. ():. 10.1007/s12033-009-9142-3 PubMed 19160076 |
买家导航
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 pSK+KanaRpsL 4678 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pSK+KanaRpsL
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4678)
AUTHORS Wang S, Zhao Y, Leiby M, Zhu J
TITLE A New Positive/Negative Selection Scheme for Precise BAC
Recombineering.
JOURNAL Mol Biotechnol. 2009 Jan 22. ():.
PUBMED 19160076
REFERENCE 2 (bases 1 to 4678)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4678
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(140..161)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 352..371
/label=F1ori-R
/note="F1 origin, reverse 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 626..645
/label=T7
/note="T7 promoter, forward primer"
promoter 626..644
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 669..685
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 670..686
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
CDS 1446..2240
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(1500..1519)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 2110..2129
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(2434..2450)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2434..2450)
/label=pBluescriptSK
/note="For pBluescript vector"
promoter complement(2487..2505)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2526..2542)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2526..2542)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2539..2561)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2550..2566
/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(2574..2604)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2619..2640
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2757..2774)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2928..3516)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3008..3027)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3687..4547)
/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 4310..4329
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4548..4652)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ggaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg cgccattcgc cattcaggct
481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tgggtaccgg
661 gccccccctc gaggtcgacg gtatcgataa gcttcacgct gccgcaagca ctcagggcgc
721 aagggctgct aaaggaagcg gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc
781 cggatgaatg tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag
841 caggtagctt gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca
901 agcgaaccgg aattgccagc tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta
961 aactggatgg ctttcttgcc gccaaggatc tgatggcgca ggggatcaag atctaaccag
1021 gagctattta atggcaacag ttaaccagct ggtacgcaaa ccacgtgctc gcaaagttgc
1081 gaaaagcaac gtgcctgcgc tggaagcatg cccgcaaaaa cgtggcgtat gtactcgtgt
1141 atatactacc actcctaaaa aaccgaactc cgcgctgcgt aaagtatgcc gtgttcgtct
1201 gactaacggt ttcgaagtga cttcctacat cggtggtgaa ggtcacaacc tgcaggagca
1261 ctccgtgatc ctgatccgtg gcggtcgtgt taaagacctc ccgggtgttc gttaccacac
1321 cgtacgtggt gcgcttgact gctccggcgt taaagaccgt aagcaggctc gttccaagta
1381 tggcgtgaag cgtcctaagg cttaatggta gatctgatca agagacagga tgacggtcgt
1441 ttcgcatgct tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
1501 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
1561 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
1621 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
1681 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
1741 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
1801 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
1861 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
1921 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
1981 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
2041 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
2101 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
2161 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
2221 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
2281 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
2341 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
2401 cttcgcccac cccgggctcg atcccctcgg ggggatccac tagttctaga gcggccgcca
2461 ccgcggtgga gctccagctt ttgttccctt tagtgagggt taattccgag cttggcgtaa
2521 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
2581 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
2641 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
2701 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
2761 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
2821 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
2881 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
2941 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
3001 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
3061 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
3121 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
3181 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
3241 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
3301 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
3361 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
3421 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
3481 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
3541 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
3601 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
3661 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
3721 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
3781 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
3841 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
3901 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
3961 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
4021 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
4081 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
4141 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
4201 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
4261 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
4321 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
4381 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
4441 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
4501 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
4561 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
4621 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctg
//
