| 质粒名称: | pAraTMDN |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 47515 |
| 存储实验室: | Bryan Berger |
|
载体骨架基本信息 |
|
| 载体名称: | pRSF |
| 空载体大小: | 6004 bp |
| 完整质粒大小: | 6001 bp |
| 载体类型: | 细菌细胞表达 |
| 载体抗性: | Kanamycin |
| 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: | Integrin alpha2B L980A TM-CYTO |
| 物种来源: | Synthetic |
| 插入大小: | 138 bp |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | SacI(未破坏) |
| 3’克隆位点: | KpnI(未破坏) |
| 5’测序引物 | gcgctgaagtcttacgagga |
| 3’测序引物 | TTATGACAACTTGACGGCTACATC |
|
文献引用方法 |
|
| 材料和方法部分: | pAraTMDN was a gift from Bryan Berger (Addgene plasmid # 47515 ; http://n2t.net/addgene:47515 ; RRID:Addgene_47515) |
| 参考文献部分: | A Novel Assay for Assessing Juxtamembrane and Transmembrane Domain Interactions Important for Receptor Heterodimerization. Su PC, Berger BW. J Mol Biol. 2013 Jul 19. pii: S0022-2836(13)00456-7. doi: 10.1016/j.jmb.2013.07.022. 10.1016/j.jmb.2013.07.022 PubMed 23876708 |
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 pAraTMDN 6001 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Contains A2B L980A TMCY mutant, expresses DN AraC (AraC*) for
heterodimer competition assay.
ACCESSION .
VERSION .
KEYWORDS pAraTMDN
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6001)
AUTHORS Su PC, Berger BW
TITLE A Novel Assay for Assessing Juxtamembrane and Transmembrane Domain
Interactions Important for Receptor Heterodimerization.
JOURNAL J Mol Biol. 2013 Jul 19. pii: S0022-2836(13)00456-7. doi:
10.1016/j.jmb.2013.07.022.
PUBMED 23876708
REFERENCE 2 (bases 1 to 6001)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6001
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 193..222
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 230..246
/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)."
CDS 385..1467
/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
YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind complement(404..423)
/label=LacI-R
/note="LacI, reverse primer"
promoter 1699..1728
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 1736..1752
/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)."
CDS 1773..2948
/codon_start=1
/gene="malE"
/product="maltose binding protein from E. coli"
/label=MBP
/note="This gene encodes a signal sequence that can direct
MBP to the bacterial periplasm."
/translation="MEIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYNG
LAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAE
ITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDK
ELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLV
DLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPS
KPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKD
PRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT"
primer_bind 2922..2945
/label=MBP-F
/note="Maltose binding protein, forward primer"
CDS 2949..2978
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
primer_bind complement(3542..3559)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 3712..3798
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3890..3917
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
CDS complement(4276..5091)
/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 4999..5018
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
promoter complement(5092..5183)
/gene="bla"
/label=AmpR promoter
rep_origin complement(5199..5948)
/direction=LEFT
/label=RSF ori
/note="Plasmids containing the RSF 1030 origin of
replication can be propagated in E. coli cells that contain
additional plasmids with compatible origins."
primer_bind complement(5209..5226)
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(5449..5468)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
241 taacaatttc acacaggaaa cagaaattcg agctcggtac ccggggatcc tctagagtcg
301 acctgcaggc atgccgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa
361 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
421 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
481 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
541 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
601 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
661 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
721 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
781 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
841 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
901 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
961 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
1021 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
1081 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
1141 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
1201 tacgacgata ccgaagacag ctcatgttat atcccgccgt caaccaccat caaacaggat
1261 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
1321 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
1381 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
1441 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agcgcgaatt
1501 gatctggttt gacagcttat catcgactgc acggtgcacc aatgcttctg gcgtcaggca
1561 gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat tcgtgtcgct
1621 caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg gttctggcaa
1681 atattctgaa atgagctgtt gacaattaat catccggctc gtataatgtg tggaattgtg
1741 agcggataac aatttcacac aggaaacaga ccatggaaat aaaaacaggt gcacgcatcc
1801 tcgcattatc cgcattaacg acgatgatgt tttccgcctc ggctctcgcc aaaatcgaag
1861 aaggtaaact ggtaatctgg attaacggcg ataaaggcta taacggtctc gctgaagtcg
1921 gtaagaaatt cgagaaagat accggaatta aagtcaccgt tgagcatccg gataaactgg
1981 aagagaaatt cccacaggtt gcggcaactg gcgatggccc tgacattatc ttctgggcac
2041 acgaccgctt tggtggctac gctcaatctg gcctgttggc tgaaatcacc ccggacaaag
2101 cgttccagga caagctgtat ccgtttacct gggatgccgt acgttacaac ggcaagctga
2161 ttgcttaccc gatcgctgtt gaagcgttat cgctgattta taacaaagat ctgctgccga
2221 acccgccaaa aacctgggaa gagatcccgg cgctggataa agaactgaaa gcgaaaggta
2281 agagcgcgct gatgttcaac ctgcaagaac cgtacttcac ctggccgctg attgctgctg
2341 acgggggtta tgcgttcaag tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg
2401 ataacgctgg cgcgaaagcg ggtctgacct tcctggttga cctgattaaa aacaaacaca
2461 tgaatgcaga caccgattac tccatcgcag aagctgcctt taataaaggc gaaacagcga
2521 tgaccatcaa cggcccgtgg gcatggtcca acatcgacac cagcaaagtg aattatggtg
2581 taacggtact gccgaccttc aagggtcaac catccaaacc gttcgttggc gtgctgagcg
2641 caggtattaa cgccgccagt ccgaacaaag agctggcaaa agagttcctc gaaaactatc
2701 tgctgactga tgaaggtctg gaagcggtta ataaagacaa accgctgggt gccgtagcgc
2761 tgaagtctta cgaggaagag ttggcgaaag atccacgtat tgccgccact atggaaaacg
2821 cccagaaagg tgaaatcatg ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc
2881 gtactgcggt gatcaacgcc gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg
2941 cgcagactga acagaaactg attagcgaag aagatctgga gctcgcgatt ccgatttggt
3001 gggtgctggt gggcgtgctg ggcggcctgc tgctggcgac cattctggtg ctggcgatgt
3061 ggaaagtggg cttttttaaa cgcaaccgcc cgccgctgga agaagatgat gaagaaggcg
3121 aaggtaccat gaacgagtcg ctccatccac cgatggataa tcgggtacgc gaggcttgtc
3181 agtacatcag cgatcacctg gcagacagca attttgatat cgccagcgtc gcacagcatg
3241 tttgcttgtc gccgtcggct ctgtcacatc ttttccgcca gcagttaggg attagcgtct
3301 taagctggcg cgaggaccaa cgtatcagcc aggcgaagct gcttttgagc accacccgga
3361 tgcctatcgc caccgtcggt cgcaatgttg gttttgacga tcaactctat ttctcgcggg
3421 tatttaaaaa atgcaccggg gccagcccga gcgagttccg tgccggttgt gaagaaaaag
3481 tgaatgatgt agccgtcaag ttgtcataag gctgttttgg cggatgagag aagattttca
3541 gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg
3601 gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg
3661 ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
3721 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
3781 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
3841 gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc
3901 ctgacggatg gcctttttgc gtttctacaa actctttttg tttatttttc taaatacatt
3961 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
4021 gactagtggt cttgaggggt tttttgctga aacctcaggc atttgagaag cacacggtca
4081 cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac
4141 gaccctgccc tgaaccgacg acaagctgac gaccgggtct ccgcaagtgg cacttttcgg
4201 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
4261 ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
4321 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
4381 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
4441 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
4501 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
4561 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
4621 attcattcgt gattgcgcct gagcgagacg aaatacgcgg tcgctgttaa aaggacaatt
4681 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
4741 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
4801 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
4861 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
4921 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
4981 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
5041 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca tactcttcct
5101 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
5161 atgtatttag aaaaataaac aaataggcat gcagcgctct tccgcttcct cgctcactga
5221 ctcgctacgc tcggtcgttc gactgcggcg agcggtgtca gctcactcaa aagcggtaat
5281 acggttatcc acagaatcag gggataaagc cggaaagaac atgtgagcaa aaagcaaagc
5341 accggaagaa gccaacgccg caggcgtttt tccataggct ccgcccccct gacgagcatc
5401 acaaaaatcg acgctcaagc cagaggtggc gaaacccgac aggactataa agataccagg
5461 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
5521 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgttggt
5581 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
5641 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
5701 acttatcgcc actggcagca gccattggta actgatttag aggactttgt cttgaagtta
5761 tgcacctgtt aaggctaaac tgaaagaaca gattttggtg agtgcggtcc tccaacccac
5821 ttaccttggt tcaaagagtt ggtagctcag cgaaccttga gaaaaccacc gttggtagcg
5881 gtggtttttc tttatttatg agatgatgaa tcaatcggtc tatcaagtca acgaacagct
5941 attccgttac tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcact
6001 a
//
