| 质粒名称: | pTP1172 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 104161 |
| 存储实验室: | Dirk Görlich |
|
载体骨架基本信息 |
|
| 载体名称: | pQE80 derivative |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 4673 bp |
| 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: | Anti-mouse kappa chain specific nanobody TP1170 (3xCysteines) |
| 物种来源: | Vicugna pacos |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: |
14xHistidine tag（主干上的N终端）
NEDD8 from Brachypodium distachyon（可被NEDP1切割）（主干上的N末端）
|
| 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: | pTP1172 was a gift from Dirk Görlich (Addgene plasmid # 104161 ; http://n2t.net/addgene:104161 ; RRID:Addgene_104161) |
| 参考文献部分: | A toolbox of anti–mouse and anti–rabbit IgG secondary nanobodies. Pleiner T, Bates M, Görlich D. J Cell Biol Dec 2017, jcb.201709115 10.1083/jcb.201709115 |
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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 pTP1172 4672 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Bacterial expression plasmid of anti-mouse kappa chain nanobody
TP1170 (3x Cysteines).
ACCESSION .
VERSION .
KEYWORDS pTP1172
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4672)
AUTHORS Pleiner T, Bates M, Gorlich D
TITLE A toolbox of anti–mouse and anti–rabbit IgG secondary nanobodies
JOURNAL J Cell Biol Dec 2017, jcb.201709115
REFERENCE 2 (bases 1 to 4672)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4672
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..45
/label=T5 promoter
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 21..37
/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)."
protein_bind 53..69
/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)."
RBS 88..99
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
terminator 908..1002
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
primer_bind complement(1050..1069)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 1061..1876
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(1134..1153)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
terminator 2095..2181
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(2274..3356)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 3318..3337
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(3357..3434)
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
primer_bind complement(3518..3540)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 3626..3766
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 3681..3700
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(3781..3798)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3952..4540)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4032..4051)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4631..4649
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taatagattc aattgtgagc
61 ggataacaat ttcacacaga attcattaaa gaggagaaat taaccatgag caagcatcac
121 catcattcag gccatcacca taccggacac caccatcatt caggcagtca tcaccatacc
181 ggcggatcat ccggaggtat gagctcagga tctaacggat ctacgggttc tggtactggt
241 aatagcggga ccggaaccat gattaaagta aagactctga ccggcaagga aattgagatc
301 gacatcgaac cgaccgacac catcgatcgt atcaaagaac gtgtggagga aaaagaaggc
361 attcctccgg ttcagcagcg tctcatttac gcgggtaaac agcttgcaga tgacaaaacc
421 gccaaagatt acaatatcga gggcggtagc gtactgcacc ttgttctggc cctgcgtggt
481 ggcgccggta cctgcggatc ccaggtgcaa ttggtagagt gtgggggagg ctgggtgcag
541 cctggggggt ctctgagact ctcctgtgca gcctctggat tcaccttcag tgacactgcc
601 atgatgtggg tccgccaggc tccagggaag gggcgagagt gggtcgcagc tattgatact
661 ggtggtggat acacatacta tgcagactcc gtgaagggcc gattcaccat ctccagagac
721 aacgccaaga acacgctgta tctgcaaatg aacagcctga aaccggagga cacggcccga
781 tactactgtg cgaagaccta ctccggtaat tactatagca actacacggt tgccaactat
841 ggcaccacgg gccgggggac cctggtcaca gtctcatcat gttaagcttc ttaattagct
901 gagcttggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
961 agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcta gccatgaaaa
1021 taaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
1081 aacgtcttgc tctaggccgc gattaaattc caacatggat gctgatttat atgggtataa
1141 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc
1201 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
1261 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
1321 tatccgtact cctgatgatg cttggttact cacgactgcg atccccggca aaacagcatt
1381 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
1441 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
1501 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
1561 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcaca aacttttgcc
1621 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
1681 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
1741 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
1801 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
1861 cgatgagttt ttctaagaat taattcatgg gcaaatatta tacgcaaggc gacaaggtgc
1921 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
1981 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag
2041 gcagttattg gtgcccttaa acgcctgggg taatgactct ctagcttgag gcatcaaata
2101 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
2161 gctctcctga gtaggacaaa tccgccctct agattacgtg cagtcgatga taagctgtca
2221 aacatgagaa ttgtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
2281 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
2341 ggagaggcgg tttgcgtatt gggggccagg gtggtttttc ttttcaccag tgagacgggc
2401 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
2461 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
2521 ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
2581 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
2641 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
2701 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
2761 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
2821 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
2881 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
2941 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
3001 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
3061 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
3121 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
3181 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
3241 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
3301 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
3361 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcac
3421 cattcgatgg tgtcggaatt tcgggcagcg ttgggtcctg gccacgggtg cgcatgatct
3481 agagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
3541 catccgctta cagacaagct gtgaccgtct ccgggagcag acaagcccgt cagggcgcgt
3601 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
3661 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
3721 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc
3781 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
3841 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
3901 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
3961 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
4021 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
4081 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
4141 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
4201 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
4261 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
4321 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
4381 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
4441 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
4501 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
4561 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgacatt
4621 aacctataaa aataggcgta tcacgaggcc ctttcgtctt cacctcgaga aa
//
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