| 质粒名称: | pWSK29 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 172972 |
| 存储实验室: | Renato Morona |
|
载体骨架基本信息 |
|
| 载体名称: | pWSK29 |
| 载体出品公司: | -- |
| 空载体大小: | 5400 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pWSK29 was a gift from Renato Morona (Addgene plasmid # 172972 ; http://n2t.net/addgene:172972 ; RRID:Addgene_172972) |
| 参考文献部分: | Altering the length of the lipopolysaccharide O antigen has an impact on the interaction of Salmonella enterica serovar Typhimurium with macrophages and complement. Murray GL, Attridge SR, Morona R. J Bacteriol. 2006 Apr;188(7):2735-9. doi: 10.1128/JB.188.7.2735-2739.2006. 10.1128/JB.188.7.2735-2739.2006 PubMed 16547065 |
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 pWSK29 5434 bp ds-DNA circular SYN 23-AUG-2021
DEFINITION Low copy number cloning vector.
ACCESSION .
VERSION .
KEYWORDS pWSK29
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5434)
AUTHORS Murray GL, Attridge SR, Morona R
TITLE Altering the length of the lipopolysaccharide O antigen has an
impact on the interaction of Salmonella enterica serovar Typhimurium
with macrophages and complement.
JOURNAL J Bacteriol. 2006 Apr;188(7):2735-9. doi:
10.1128/JB.188.7.2735-2739.2006.
PUBMED 16547065
REFERENCE 2 (bases 1 to 5434)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 23, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5434
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(1297..2247)
/codon_start=1
/gene="rep101"
/product="RepA protein needed for replication with the
pSC101 origin"
/label=Rep101
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
rep_origin 2295..2517
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
primer_bind 3662..3679
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 3796..3817
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3832..3862
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3870..3886
/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 3875..3897
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3894..3910
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3894..3910
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 3906..4481
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
TDRPSQQLRSLNGEWDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG
FPRQALNRGLPLGFRFSALRHLDPKKLD"
primer_bind 3929..3949
/label=T3
/note="T3 promoter, forward primer"
promoter 3931..3949
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
misc_feature 3962..4069
/label=MCS
/note="pBluescript multiple cloning site"
primer_bind 3986..4002
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3986..4002
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(4036..4052)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4037..4053)
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind complement(4077..4096)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4078..4096)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4106..4123)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4106..4122)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4115..4137)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 4264..4719
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4351..4370)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4561..4582
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 4745..4849
/gene="bla"
/label=AmpR promoter
CDS 4850..276
/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 complement(5068..5087)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
ORIGIN
1 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
61 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
121 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
181 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
241 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
301 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
361 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagat
421 ctctgttcgg gggttcactg acgctgcgtg actacgatgg tcaggttgcg gaggctatgg
481 ccctggtacg agcgctgaac aaaatgacga aagcaggtat gcctgaaagc gtgcgtattg
541 cctgaaaaca caacccgcta cgggggagac ttacccgaaa tctgatttat tcaacaaagc
601 cactttcagt atggatttgg gtgttacggg ggggggttgg tgcttaaaat cgcgctgagg
661 atttctgatg gtgttaagcg ggcggttttg agatgtaaac tcgcccgttt aacataatgg
721 atcttgcgcg caccgcccga acaccactcg ccacaaaaaa ccgccggaac gtccaaaagt
781 acgggttttg ctgcccgcaa acgggctgtt ctggtgttgc tagtttgtta tcagaatcgc
841 agatccggct tcagccggtt tgccggctga aagcgctatt tcttccagaa ttgccatgat
901 tttttcccca cgggaggcgt cactggctcc cgtgttgtcg gcagctttga ttcgataagc
961 agcatcgcct gtttcaggct gtctatgtgt gactgttgag ctgtaacaag ttgtctcagg
1021 tgttcaattt catgttctag ttgctttgtt ttactggttt cacctgttct attaggtgtt
1081 acatgctgtt catctgttac attgtcgatc tgttcatggt gaacagcttt gaatgcacca
1141 aaaactcgta aaagctctga tgtatctatc ttttttacac cgttttcatc tgtgcatatg
1201 gacagttttc cctttgatat gtaacggtga acagttgttc tacttttgtt tgttagtctt
1261 gatgcttcac tgatagatac aagagccata agaacctcag atccttccgt atttagccag
1321 tatgttctct agtgtggttc gttgtttttg cgtgagccat gagaacgaac cattgagatc
1381 atacttactt tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc
1441 agttaaagca tcgtgtagtg tttttcttag tccgttatgt aggtaggaat ctgatgtaat
1501 ggttgttggt attttgtcac cattcatttt tatctggttg ttctcaagtt cggttacgag
1561 atccatttgt ctatctagtt caacttggaa aatcaacgta tcagtcgggc ggcctcgctt
1621 atcaaccacc aatttcatat tgctgtaagt gtttaaatct ttacttattg gtttcaaaac
1681 ccattggtta agccttttaa actcatggta gttattttca agcattaaca tgaacttaaa
1741 ttcatcaagg ctaatctcta tatttgcctt gtgagttttc ttttgtgtta gttcttttaa
1801 taaccactca taaatcctca tagagtattt gttttcaaaa gacttaacat gttccagatt
1861 atattttatg aattttttta actggaaaag ataaggcaat atctcttcac taaaaactaa
1921 ttctaatttt tcgcttgaga acttggcata gtttgtccac tggaaaatct caaagccttt
1981 aaccaaagga ttcctgattt ccacagttct cgtcatcagc tctctggttg ctttagctaa
2041 tacaccataa gcattttccc tactgatgtt catcatctga gcgtattggt tataagtgaa
2101 cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca
2161 gcataaaatt agcttggttt catgctccgt taagtcatag cgactaatcg ctagttcatt
2221 tgctttgaaa acaactaatt cagacataca tctcaattgg tctaggtgat tttaatcact
2281 ataccaattg agatgggcta gtcaatgata attactagtc cttttccttt gagttgtggg
2341 tatctgtaaa ttctgctaga cctttgctgg aaaacttgta aattctgcta gaccctctgt
2401 aaattccgct agacctttgt gtgttttttt tgtttatatt caagtggtta taatttatag
2461 aataaagaaa gaataaaaaa agataaaaag aatagatccc agccctgtgt ataactcact
2521 actttagtca gttccgcagt attacaaaag gatgtcgcaa acgctgtttg ctcctctaca
2581 aaacagacct taaaacccta aaggcttaag tagcaccctc gcaagctcgg gcaaatcgct
2641 gaatattcct tttgtctccg accatcaggc acctgagtcg ctgtcttttt cgtgacattc
2701 agttcgctgc gctcacggct ctggcagtga atgggggtaa atggcactac aggcgccttt
2761 tatggattca tgcaaggaaa ctacccataa tacaagaaaa gcccgtcacg ggcttctcag
2821 ggcgttttat ggcgggtctg cggagtggtg aatccgttag cgaggtgccg ccggcttcca
2881 ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt
2941 atagggcggc gcctacaatc catgccaacc cgttccatgt gctcgccgag gcggcataaa
3001 tcgccgtgac gatcagcggt ccaatgatcg aagttaggct ggtaagagcc gcgagcgatc
3061 cttgaagctg tccctgatgg tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg
3121 gcatcccgat gccgccggaa gcgagaagaa tcataatggg gaaggccatc cagcctcgcg
3181 tcgcgaacgc cagcaagacg tagcccagcg cgtcggccgc catgccggcg ataatggcct
3241 gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca
3301 agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct
3361 cgccgaaaat gacccagagc gctgccggca ccctgtccta cgagttgcat gataaagaag
3421 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
3481 ttgaaggctc tcaagggcat cggcattcgc cattcaggct gcctcctacc aagacgactt
3541 caaatccgac ccgtagagat ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
3601 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
3661 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
3721 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
3781 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
3841 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
3901 cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct
3961 ggagctccac cgcggtggcg gccgctctag aactagtgga tcccccgggc tgcaggaatt
4021 cgatatcaag cttatcgata ccgtcgacct cgaggggggg cccggtaccc aattcgccct
4081 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
4141 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
4201 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
4261 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
4321 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
4381 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
4441 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
4501 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
4561 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
4621 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
4681 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
4741 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
4801 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
4861 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
4921 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
4981 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
5041 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
5101 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
5161 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
5221 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
5281 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgctggga
5341 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
5401 ggcaacaacg ttgcgcaaac tattaactgg cgaa
//
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