| 质粒名称: | pAA3.stelN |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 112011 |
| 存储实验室: | Stephen Harrison |
|
载体骨架基本信息 |
|
| 载体名称: | pAA.3 |
| 载体类型: | 细菌表达 |
| 载体抗性: | 氨苄霉素 |
| 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: | TelN |
| 插入大小: | 2043 bp |
| 启动子: | T7 |
| 标签/融合蛋白: | strep和his（插入件上的N端子） |
| 克隆方法: | Gibson克隆 |
| 5’测序引物 | TCTAGACTTCATTTCCTTCT |
| 3’测序引物 | TTATGTAAATAAGTTGGCCA |
|
文献引用方法 |
|
| 材料和方法部分: | pAA3.stelN was a gift from Stephen Harrison (Addgene plasmid # 112011 ; http://n2t.net/addgene:112011 ; RRID:Addgene_112011) |
| 参考文献部分: | Structure of the DASH/Dam1 complex shows its role at the yeast kinetochore-microtubule interface. Jenni S, Harrison SC. Science. 2018 May 4;360(6388):552-558. doi: 10.1126/science.aar6436. 10.1126/science.aar6436 PubMed 29724956 |
买家导航
pAA3stelN载体基本信息
pAA3stelN载体图谱质粒图谱和多克隆位点信息
pAA3stelN载体简介
pAA3stelN载体序列
LOCUS pAA3.stelN 4639 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses TelN enzyme.
ACCESSION .
VERSION .
KEYWORDS pAA3.stelN
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4639)
AUTHORS Jenni S, Harrison SC
TITLE Structure of the DASH/Dam1 complex shows its role at the yeast
kinetochore-microtubule interface.
JOURNAL Science. 2018 May 4;360(6388):552-558. doi: 10.1126/science.aar6436.
PUBMED 29724956
REFERENCE 2 (bases 1 to 4639)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4639
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind complement(138..171)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
primer_bind 361..380
/label=T7
/note="T7 promoter, forward primer"
promoter 361..379
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 411..433
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 453..476
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 492..515
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 531..554
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS 555..572
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 573..593
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 594..2486
/codon_start=1
/gene="telN"
/product="protelomerase of coliphage N15"
/label=TelN
/note="generates the covalently closed hairpin telomeres"
/translation="SKVKIGELINTLVNEVEAIDASDRPQGDKTKRIKAAAARYKNALF
NDKRKFRGKGLQKRITANTFNAYMSRARKRFDDKLHHSFDKNINKLSEKYPLYSEELSS
WLSMPTANIRQHMSSLQSKLKEIMPLAEELSNVRIGSKGSDAKIARLIKKYPDWSFALS
DLNSDDWKERRDYLYKLFQQGSALLEELHQLKVNHEVLYHLQLSPAERTSIQQRWADVL
REKKRNVVVIDYPTYMQSIYDILNNPATLFSLNTRSGMAPLAFALAAVSGRRMIEIMFQ
GEFAVSGKYTVNFSGQAKKRSEDKSVTRTIYTLCEAKLFVELLTELRSCSAASDFDEVV
KGYGKDDTRSENGRINAILAKAFNPWVKSFFGDDRRVYKDSRAIYARIAYEMFFRVDPR
WKNVDEDVFFMEILGHDDENTQLHYKQFKLANFSRTWRPEVGDENTRLVALQKLDDEMP
GFARGDAGVRLHETVKQLVEQDPSAKITNSTLRAFKFSPTMISRYLEFAADALGQFVGE
NGQWQLKIETPAIVLPDEESVETIDEPDDESQDDELDEDEIELDEGGGDEPTEEEGPEE
HQPTALKPVFKPAKNNGDGTYKIEFEYDGKHYAWSGPADSPMAAMRSAWETYYS"
primer_bind complement(2558..2576)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2572..2619
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind complement(2682..2704)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 2790..2929
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 2845..2864
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(2944..2961)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3115..3703)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3195..3214)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
terminator 3734..3773
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
promoter complement(4638..90)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
61 tatttgaatg tatttagaaa aataaacaaa taggggaatt aaaaaaaagc ccgctcatta
121 ggcgggctac tagtcagata acttcgtata atgtatgcta tacgaagtta tggtaccgcg
181 gccgcgtaga ggatctgttg atcagcagtt caacctgttg atagtacttc gttaatacag
241 atgtaggtgt tggcaccatg cataactata acggtcctaa ggtagcgacc taggttctag
301 acttcatttc cttctccaga ttaactcaat tccagcattt agaaccctac caagtatcga
361 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
421 tttaagaagg agatatacat atgaaatctt cttggagcca tccgcagttt gaaaaaggcg
481 ccatgaccgg ttggtcccac ccgcagttcg aaaaacgtag cgcgggcagc tggagtcatc
541 cgcaattcga aaaacaccat caccatcacc atgaaaacct gtacttccaa tccagcaagg
601 taaaaatcgg tgagttgatc aacacgcttg tgaatgaggt agaggcaatt gatgcctcag
661 accgcccaca aggcgacaaa acgaagagaa ttaaagccgc agccgcacgg tataagaacg
721 cgttatttaa tgataaaaga aagttccgtg ggaaaggatt gcagaaaaga ataaccgcga
781 atacttttaa cgcctatatg agcagggcaa gaaagcggtt tgatgataaa ttacatcata
841 gctttgataa aaatattaat aaattatcgg aaaagtatcc tctttacagc gaagaattat
901 cttcatggct ttctatgcct acggctaata ttcgccagca catgtcatcg ttacaatcta
961 aattgaaaga aataatgccg cttgccgaag agttatcaaa tgtaagaata ggctctaaag
1021 gcagtgatgc aaaaatagca agactaataa aaaaatatcc agattggagt tttgctctta
1081 gtgatttaaa cagtgatgat tggaaggagc gccgtgacta tctttataag ttattccaac
1141 aaggctctgc gttgttagaa gaactacacc agctcaaggt caaccatgag gttctgtacc
1201 atctgcagct aagccctgcg gagcgtacat ctatacagca acgatgggcc gatgttctgc
1261 gcgagaagaa gcgtaatgtt gtggttattg actacccaac atacatgcag tctatctatg
1321 atattttgaa taatcctgcg actttattta gtttaaacac tcgttctgga atggcacctt
1381 tggcctttgc tctggctgcg gtatcagggc gaagaatgat tgagataatg tttcagggtg
1441 aatttgccgt ttcaggaaag tatacggtta atttctcagg gcaagctaaa aaacgctctg
1501 aagataaaag cgtaaccaga acgatttata ctttatgcga agcaaaatta ttcgttgaat
1561 tattaacaga attgcgttct tgctctgctg catctgattt cgatgaggtt gttaaaggat
1621 atggaaagga tgatacaagg tctgagaacg gcaggataaa tgctatttta gcaaaagcat
1681 ttaacccttg ggttaaatca tttttcggcg atgaccgtcg tgtttataaa gatagccgcg
1741 ctatttacgc tcgcatcgct tatgagatgt tcttccgcgt cgatccacgg tggaaaaacg
1801 tcgacgagga tgtgttcttc atggagattc tcggacacga cgatgagaac acccagctgc
1861 actataagca gttcaagctg gccaacttct ccagaacctg gcgacctgaa gttggggatg
1921 aaaacaccag gctggtggct ctgcagaaac tggacgatga aatgccaggc tttgccagag
1981 gtgacgctgg cgtccgtctc catgaaaccg ttaagcagct ggtggagcag gacccatcag
2041 caaaaataac caacagcact ctccgggcct ttaaatttag cccgacgatg attagccggt
2101 acctggagtt tgccgctgat gcattggggc agttcgttgg cgagaacggg cagtggcagc
2161 tgaagataga gacacctgca atcgtcctgc ctgatgaaga atccgttgag accatcgacg
2221 aaccggatga tgagtcccaa gacgacgagc tggatgaaga tgaaattgag ctcgacgagg
2281 gtggcggcga tgaaccaacc gaagaggaag ggccagaaga acatcagcca actgctctaa
2341 aacccgtctt caagcctgca aaaaataacg gggacggaac gtacaagata gagtttgaat
2401 acgatggaaa gcattatgcc tggtccggcc ccgccgatag ccctatggcc gcaatgcgat
2461 ccgcatggga aacgtactac agctaactcg agagatccgg ctgctaacaa acccgcttaa
2521 ggaagatggc caacttattt acataaggct gctgccaccg ctgagcaata actagcataa
2581 ccccttgggg cctctaaacg ggtcttgagg ggttttttgg tttaaaccca tctaattggg
2641 aattccccct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
2701 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
2761 gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc
2821 ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccatt
2881 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc
2941 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
3001 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
3061 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
3121 taggttccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
3181 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
3241 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
3301 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
3361 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
3421 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
3481 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
3541 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
3601 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
3661 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
3721 ttctaccggt ccgtccaaaa aaaaaggctc caaaaggagc ctttaattgt atcggtttac
3781 caatgcttaa tcagtgaagc ccctatttca gcaatctgtc tatttcgttc gtccatagtt
3841 gcctgactac ccgtcgtgta gataactacg atacgggaag gcttaccatc tggtccaagt
3901 gcggcaatga taccccgtga accacgctct cctgccccag atttatcagc aataaaccag
3961 ccagcaggaa gtgccgaacg cagtagtggt cctgcaactt tatccgcttc aagccagtct
4021 attaattgtt gccgtgaagc cagagtaagt agttcgccag ttaatagttt gcgtaatgtt
4081 gttgccattg ccgcaggcat tgtggtgtct cgctcgtcgt ttggtatggc ttcattcagt
4141 tctggttccc aacggtcaag gcgagttaca tggtctccca tattgtgcaa aaaagcggtt
4201 aattccttcg gtccaccgat ggttgtcaga agtaagttcg ccgcagtgtt atcactcatc
4261 gttatggcgg cactacataa ttctcttacc gtcattccgt ccgtaaggtg cttttctgtc
4321 actggggagt attcaaccaa gtcattctgc gaatagtgta ttcgtcgtcc gagttgctct
4381 tgccccgcat ctacacggga taatactgct ccacatagca gaactttgaa agtgctcatc
4441 attggaaaac gctcttcagg gcgaaaactc tcaaggattt taccgctatt caagtccagt
4501 tctatgtatc ccactcgtgc ccccaattgg tcctctgcgt cttttacttt cacgagcgtt
4561 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
4621 aaatgttgaa tactcatac
//
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