| 质粒名称: | pDEST40-2XFL-wtSrc |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 140295 |
| 存储实验室: | Mark Moasser |
|
载体骨架基本信息 |
|
| 载体名称: | modified pcDNA-pDEST40 |
| 载体出品公司: | Clontech |
| 空载体大小: | 7100bp |
| 完整质粒大小: | 7175bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 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: | Src wildtype、S4-UD-S32-KD |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1608bp |
| 标签/融合蛋白: | 2X Flag (C terminal on backbone) |
| 克隆方法: | Gateway Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | pDEST40-2XFL-wtSrc was a gift from Mark Moasser (Addgene plasmid # 140295 ; http://n2t.net/addgene:140295 ; RRID:Addgene_140295) |
| 参考文献部分: | A Dimerization Function in the Intrinsically Disordered N-Terminal Region of Src. Spassov DS, Ruiz-Saenz A, Piple A, Moasser MM. Cell Rep. 2018 Oct 9;25(2):449-463.e4. doi: 10.1016/j.celrep.2018.09.035. 10.1016/j.celrep.2018.09.035 PubMed 30304684 |
买家导航
pDEST40-2XFL-wtSrc载体质粒基本信息
pDEST40-2XFL-wtSrc质粒图谱载体图谱和pDEST40-2XFL-wtSrc载体序列质粒序列多克隆位点信息
pDEST40-2XFL-wtSrc质粒载体简介
pDEST40-2XFL-wtSrc质粒序列载体序列
LOCUS pDEST40-2XFL-wtSrc 7175 bp DNA circular SYN 14-APR-2025
DEFINITION Expresses Flag-tagged wild-type human Src in mammalian cells for
structure-function studies.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7175)
AUTHORS Spassov DS, Ruiz-Saenz A, Piple A, Moasser MM
TITLE A Dimerization Function in the Intrinsically Disordered N-Terminal
Region of Src.
JOURNAL Cell Rep. 2018 Oct 9;25(2):449-463.e4. doi:
10.1016/j.celrep.2018.09.035.
PUBMED 30304684
REFERENCE 2 (bases 1 to 7175)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.celrep.2018.09"; journalName: "Cell Rep"; date:
"2018-10-9- 9"; volume: "25"; issue: "2"; pages: "449-463.e4"
COMMENT Sequence Label: pDEST40-2XFL-wtSrc
FEATURES Location/Qualifiers
source 1..7175
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 911..935
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(2562..2586)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 2597..2620
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 2624..2647
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 2673..2714
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 2724..2741
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2764..2781)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2770..2994
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3040..3468
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3127..3146)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3337..3358
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3477..3497)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3482..3811
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3662..3797
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3724..3743
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3878..4672
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3932..3951)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4542..4561
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4848..4969
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4885..4904)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4939..4958
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5018..5034)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5018..5034)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5031..5053)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5042..5058
/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)."
promoter complement(join(5066..5072,5073..5090,5091..5096))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5111..5132
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5249..5266)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5420..6008)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5500..5519)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(6179..6970,6971..7039))
/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 6802..6821
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7040..7144)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
901 taagctatca acaagtttgt acaaaaaagc aggcttcgaa ggagatagaa ccatgggtag
961 caacaagagc aagcccaagg atgccagcca gcggcgccgc agcctggagc ccgccgagaa
1021 cgtgcacggc gctggcgggg gcgctttccc cgcctcgcag acccccagca agccagcctc
1081 ggccgacggc caccgcggcc ccagcgcggc cttcgccccc gcggccgccg agcccaagct
1141 gttcggaggc ttcaactcct cggacaccgt cacctccccg cagagggcgg gcccgctggc
1201 cggtggagtg accacctttg tggccctcta tgactatgag tctaggacgg agacagacct
1261 gtccttcaag aaaggcgagc ggctccagat tgtcaacaac acagagggag actggtggct
1321 ggcccactcg ctcagcacag gacagacagg ctacatcccc agcaactacg tggcgccctc
1381 cgactccatc caggctgagg agtggtattt tggcaagatc accagacggg agtcagagcg
1441 gttactgctc aatgcagaga acccgagagg gaccttcctc gtgcgagaaa gtgagaccac
1501 gaaaggtgcc tactgcctct cagtgtctga cttcgacaac gccaagggcc tcaacgtgaa
1561 gcactacaag atccgcaagc tggacagcgg cggcttctac atcacctccc gcacccagtt
1621 caacagcctg cagcagctgg tggcctacta ctccaaacac gccgatggcc tgtgccaccg
1681 cctcaccacc gtgtgcccca cgtccaagcc gcagactcag ggcctggcca aggatgcctg
1741 ggagatccct cgggagtcgc tgcggctgga ggtcaagctg ggccagggct gctttggcga
1801 ggtgtggatg gggacctgga acggtaccac cagggtggcc atcaaaaccc tgaagcctgg
1861 cacgatgtct ccagaggcct tcctgcagga ggcccaggtc atgaagaagc tgaggcatga
1921 gaagctggtg cagttgtatg ctgtggtttc agaggagccc atttacatcg tcacggagta
1981 catgagcaag gggagtttgc tggactttct caagggggag acaggcaagt acctgcggct
2041 gcctcagctg gtggacatgg ctgctcagat cgcctcaggc atggcgtacg tggagcggat
2101 gaactacgtc caccgggacc ttcgtgcagc caacatcctg gtgggagaga acctggtgtg
2161 caaagtggcc gactttgggc tggctcggct cattgaagac aatgagtaca cggcgcggca
2221 aggtgccaaa ttccccatca agtggacggc tccagaagct gccctctatg gccgcttcac
2281 catcaagtcg gacgtgtggt ccttcgggat cctgctgact gagctcacca caaagggacg
2341 ggtgccctac cctgggatgg tgaaccgcga ggtgctggac caggtggagc ggggctaccg
2401 gatgccctgc ccgccggagt gtcccgagtc cctgcacgac ctcatgtgcc agtgctggcg
2461 gaaggagcct gaggagcggc ccaccttcga gtacctgcag gccttcctgg aggactactt
2521 cacgtccacc gagccccagt accagcccgg ggagaacctc gacccagctt tcttgtacaa
2581 agtggttgat ctagaggatt acaaggatga cgacgataag ggagattaca aggatgacga
2641 cgataagtga tctagagggc ccgcggttcg aaggtaagcc tatccctaac cctctcctcg
2701 gtctcgattc tacgcgtacc ggtcatcatc accatcacca ttgagtttaa acccgctgat
2761 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
2821 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
2881 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
2941 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
3001 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
3061 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
3121 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
3181 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
3241 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
3301 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
3361 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
3421 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
3481 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
3541 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
3601 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
3661 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
3721 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
3781 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
3841 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
3901 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
3961 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
4021 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
4081 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
4141 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
4201 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
4261 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
4321 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
4381 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
4441 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
4501 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
4561 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
4621 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
4681 ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
4741 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
4801 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
4861 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
4921 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
4981 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
5041 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
5101 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
5161 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
5221 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
5281 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
5341 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
5401 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
5461 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
5521 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
5581 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
5641 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
5701 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
5761 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
5821 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
5881 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
5941 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
6001 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
6061 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
6121 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
6181 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
6241 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
6301 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
6361 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
6421 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
6481 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
6541 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
6601 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
6661 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
6721 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
6781 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
6841 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
6901 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
6961 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
7021 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
7081 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
7141 cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
