| 质粒名称: | TBXTA-c027 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 139764 |
| 存储实验室: | Opher Gileadi |
|
载体骨架基本信息 |
|
| 载体名称: | pHGT-Bio (GenBank: MH763823.1) |
| 载体出品公司: | SGC |
| 空载体大小: | 6080 bp |
| 完整质粒大小: | 7392 bp |
| 载体修饰: |
-- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 卡那霉素，50μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | TBXT, Full-length, G177D |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1340 bp |
| 基因突变: | G177D；仅含氨基酸S2-M435 |
| GenBank编号 | -- |
| 启动子: | T7 |
| 标签/融合蛋白: |
His6 GST TEV（主干上的N端）Avi标签（生物素）（主干上的C端）
|
| 克隆方法: | Ligation Independent Cloning |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | pGEX5: GGGCTGGCAAGCCACGTTTGGTG |
| 3’测序引物 | T7R |
|
文献引用方法 |
|
| 材料和方法部分: | TBXTA-c027 was a gift from Opher Gileadi (Addgene plasmid # 139764 ; http://n2t.net/addgene:139764 ; RRID:Addgene_139764) |
| 参考文献部分: | -- |
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 TBXTA-c027 7392 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Protein expression/Streptavidin pull-down.
ACCESSION .
VERSION .
KEYWORDS TBXTA-c027
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7392)
TITLE Opher Gileadi SGC expression vectors
REFERENCE 2 (bases 1 to 7392)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7392
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 1..20
/label=T7
/note="T7 promoter, forward primer"
promoter 1..19
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 20..44
/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 59..81
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 92..109
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 116..769
/codon_start=1
/product="glutathione S-transferase from Schistosoma
japonicum"
/label=GST
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGADHPPK"
primer_bind 727..749
/label=pGEX 5'
/note="pGEX vectors, Glutathione-S-transferase, forward
primer"
CDS 797..817
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 2147..2191
/codon_start=1
/product="peptide tag that allows for enzymatic
biotinylation"
/label=AviTag(TM)
/translation="GLNDIFEAQKIEWHE"
CDS 2257..2274
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2327..2345)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2341..2388
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 2425..2880
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2512..2531)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2722..2743
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(2972..3787)
/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 3695..3714
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 3779..3798
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 3909..4497
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4398..4417
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4651..4668
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 4683..4822
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(4748..4767)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4908..4930
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(4924..5115)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(5924..7006)
/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 6968..6987
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(7007..7084)
/gene="lacI"
/label=lacI promoter
primer_bind 7290..7309
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
61 tgtttaactt taagaaggag atatacatat gcaccatcat catcatcatt cttctatgtc
121 ccctatacta ggttattgga aaattaaggg ccttgtgcaa cccactcgac ttcttttgga
181 atatcttgaa gaaaaatatg aagagcattt gtatgagcgc gatgaaggtg ataaatggcg
241 aaacaaaaag tttgaattgg gtttggagtt tcccaatctt ccttattata ttgatggtga
301 tgttaaatta acacagtcta tggccatcat acgttatata gctgacaagc acaacatgtt
361 gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt gaaggagcgg ttttggatat
421 tagatacggt gtttcgagaa ttgcatatag taaagacttt gaaactctca aagttgattt
481 tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat cgtttatgtc ataaaacata
541 tttaaatggt gatcatgtaa cccatcctga cttcatgttg tatgacgctc ttgatgttgt
601 tttatacatg gacccaatgt gcctggatgc gttcccaaaa ttagtttgtt ttaaaaaacg
661 tattgaagct atcccacaaa ttgataagta cttgaaatcc agcaagtata tagcatggcc
721 tttgcagggc tggcaagcca cgtttggtgg tgccgaccat cctccaaaat cgagctcagg
781 tgtagatctg ggtaccgaga acctgtactt ccaatccatg tccaatagca gccctggcac
841 cgaaagcgcc ggtaagagcc tacagtaccg tgtggaccat ttactgagcg ccgtggagaa
901 cgaactacag gcaggcagcg aaaaaggtga cccgaccgaa cgcgaactgc gtgtgggtct
961 ggaagaaagc gaactgtggc tgcgctttaa agagctgacc aacgaaatga tcgtgaccaa
1021 aaacggccgc cgcatgtttc ctgtgctgaa agttaacgtg agcggcctgg acccgaacgc
1081 catgtacagc ttcctgctgg atttcgttgc cgcagataac caccgctgga agtatgtgaa
1141 cggcgaatgg gtgccgggtg gcaaaccgga accgcaggca cctagctgcg tgtatatcca
1201 cccggatagc cctaacttcg gtgcccattg gatgaaagcc ccggtgagct tcagcaaggt
1261 gaagctgacc aacaaactga acggcggcgg ccaaatcatg ctgaacagct tacataaata
1321 tgaacctcgc atccacattg tgcgtgtggg cgacccgcaa cgtatgatca ccagccattg
1381 cttcccggag acacagttca ttgccgttac cgcctaccag aacgaggaaa ttaccgcact
1441 gaaaattaaa tacaaccctt ttgcaaaagc cttcttagat gccaaagagc gcagcgacca
1501 caaagaaatg atggaggaac cgggcgacag ccagcagcct ggttatagcc aatggggctg
1561 gctgctgccg ggtaccagta cactgtgccc gccggcaaac ccgcatcctc agtttggtgg
1621 cgccctgagc ttaccgagca cccacagttg tgatcgctat cctaccctgc gtagccatcg
1681 tagcagtccg tacccgagcc cgtatgcaca tcgtaacaac agcccgacat atagcgacaa
1741 cagcccggcc tgtctgagca tgctacagag ccatgataac tggagcagcc tgggcatgcc
1801 ggcacatccg agcatgctgc cggtgagtca caatgccagc ccgccgacca gtagcagcca
1861 gtatcctagc ctgtggagcg ttagtaacgg tgcagtgacc ccgggtagcc aggcagcagc
1921 agttagcaat ggtctgggcg cccagttttt tcgcggcagt ccggcacatt acaccccgct
1981 gacccatcct gtgagtgccc cgagcagtag tggtagcccg ctgtatgaag gtgccgcagc
2041 cgccaccgac attgttgaca gccagtatga tgccgccgca cagggtcgcc tgattgcaag
2101 ctggacccct gtgagtccgc cgagcatgag cagtaaaggt ggatacggcc tgaatgatat
2161 ctttgaagcg cagaagattg aatggcatga atgacagtaa aggtggatac ggatccgaat
2221 tcgagctccg tcgacaagct tgcggccgca ctcgagcacc accaccacca ccactgagat
2281 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
2341 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
2401 actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
2461 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
2521 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
2581 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
2641 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
2701 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
2761 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
2821 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
2881 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
2941 ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa
3001 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
3061 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
3121 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
3181 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat
3241 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
3301 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
3361 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
3421 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
3481 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
3541 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
3601 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
3661 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
3721 aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat
3781 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc
3841 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
3901 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
3961 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
4021 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
4081 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
4141 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
4201 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
4261 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
4321 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
4381 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
4441 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
4501 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
4561 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
4621 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
4681 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
4741 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
4801 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
4861 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
4921 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
4981 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
5041 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
5101 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
5161 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
5221 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
5281 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
5341 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
5401 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
5461 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
5521 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
5581 tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg tggcgggacc
5641 agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg acaggccgat
5701 catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg ctgccggcac
5761 ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc
5821 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc
5881 cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca
5941 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
6001 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat
6061 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca
6121 gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg
6181 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg
6241 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc
6301 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc
6361 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca
6421 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg
6481 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga
6541 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca
6601 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg
6661 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca
6721 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg
6781 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca
6841 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt
6901 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag
6961 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga
7021 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg
7081 tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
7141 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
7201 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
7261 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
7321 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
7381 atcccgcgaa at
//
