| 质粒名称: | pRK GFP Paxillin |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 50529 |
| 存储实验室: | Kenneth Yamada |
|
载体骨架基本信息 |
|
| 载体名称: | pGZ21dxZ |
| 载体出品公司: | Yamada Lab, Tamura et al., Science 280: 1614-7 (1998) |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | Paxillin,PXN,focal adhesion protein, mutated |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | CMV |
| 标签/融合蛋白: | GFP21（主干上的N终端） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | HindIII (未破坏) |
| 3’克隆位点: | XbaI (未破坏) |
| 5’测序引物 | GFP FW (AAAGACCCCAACGAGAAGCG) |
| 3’测序引物 | GFP ASO (TTGTAACCATTATAAGCTGC) |
|
文献引用方法 |
|
| 材料和方法部分: | pRK GFP Paxillin was a gift from Kenneth Yamada (Addgene plasmid # 50529 ; http://n2t.net/addgene:50529 ; RRID:Addgene_50529) |
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 pRK GFP Paxillin 7075 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION This plasmid contains GFP21, which has a sequence similar to YFP,
but emission/excitation similar to GFP..
ACCESSION .
VERSION .
KEYWORDS pRK GFP Paxillin
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7075)
AUTHORS Yamada KM
TITLE Yamada Lab Plasmids
JOURNAL unpublished
REFERENCE 2 (bases 1 to 7075)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7075
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 1..589
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 490..509
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 743..760
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 877..898
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 913..943
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 951..967
/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)."
primer_bind 956..978
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 975..991
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 975..991
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
enhancer 1020..1399
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1400..1603
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 1554..1574
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 1600..1624
/label=LNCX
/note="Human CMV promoter, forward primer"
promoter 1834..1852
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 1834..1851
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind 1865..1884
/label=pMT2-F
/note="Synthetic intron, forward primer"
regulatory 1930..1939
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1936..2652
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1981..2002)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2242..2261)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2589..2610
/label=EGFP-C
/note="EGFP, forward primer"
regulatory 4381..4390
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
polyA_signal 4394..4528
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4431..4450)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4485..4504
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
promoter 4597..4954
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(4620..4640)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 4805..4940
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4867..4886
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(4974..4991)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4974..4990)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4983..5005)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 5203..5658
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5290..5309)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5500..5521
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(5675..5694)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5794..5816
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5854..5872)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5940..6044
/gene="bla"
/label=AmpR promoter
CDS 6045..6905
/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 complement(6263..6282)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
ORIGIN
1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
961 ataacaattt cacacaggaa acagctatga catgattacg aattaattcg agctcgcccg
1021 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
1081 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
1141 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
1201 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
1261 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
1321 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
1381 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
1441 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
1501 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
1561 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
1621 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc
1681 cagcctccgc ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca agagtgacgt
1741 aagtaccgcc tatagagtct ataggcccac ccccttggct tcgttagaac gcggctacaa
1801 ttaatacata accttatgta tcatacacat acgatttagg tgacactata gaataacatc
1861 cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcacct cggttctatc
1921 gattcagccg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
1981 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
2041 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
2101 gtgccctggc ccaccctcgt gaccaccttc gcctacggcc tgcagtgctt cgcccgctac
2161 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
2221 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
2281 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
2341 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
2401 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
2461 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
2521 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
2581 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
2641 gagctgtaca agggaggcgg atccaagctt atggacgacc tcgacgccct gctggcggac
2701 ttggagtcta ccacctccca catctccaaa cggcctgtgt tcttgtcgga ggagaccccc
2761 tactcatacc caactggaaa ccacacatac caggagattg ccgtgccacc ccccgtcccc
2821 ccacccccgt ccagcgaggc cctcaatggc acaatccttg accccttaga ccagtggcag
2881 cccagcggct cccgattcat ccaccagcag cctcagtcct catcacctgt gtacggctcc
2941 agtgccaaaa cttccagtgt ctccaaccct caggacagtg ttggctctcc gtgctcccga
3001 gtgggtgagg aggagcacgt ctacagcttc cccaacaagc agaaatcagc tgagccttca
3061 cccaccgtaa tgagcacgtc cctgggcagc aacctttctg aactcgaccg cctgctgctg
3121 gaactgaacg ctgtacagca taacccgcca ggcttccctg cagatgaggc caactcaagc
3181 cccccgcttc ctggggccct gagccccctc tatggtgtcc cagagactaa cagccccttg
3241 ggaggcaaag ctgggcccct gacgaaagag aagcctaagc ggaatggggg ccggggcctg
3301 gaggacgtgc ggcccagtgt ggagagtctc ttggatgaac tggagagctc cgtgcccagc
3361 cccgtccctg ccatcactgt gaaccagggc gagatgagca gcccgcagcg cgtcacctcc
3421 acccaacagc agacacgcat ctcggcctcc tctgccacca gggagctgga cgagctgatg
3481 gcttcgctgt cggatttcaa gttcatggcc caggggaaga cagggagcag ctcaccccct
3541 ggggggcccc cgaagcccgg gagccagctg gacagcatgc tggggagcct gcagtctgac
3601 ctgaacaagc tgggggtcgc cacagtcgcc aaaggagtct gcggggcctg caagaagccc
3661 atcgccgggc aggttgtgac cgccatgggg aagacgtggc accccgagca cttcgtctgc
3721 acccactgcc aggaggagat cggatcccgg aacttcttcg agcgggatgg acagccctac
3781 tgtgaaaagg actaccacaa cctcttctcc ccgcgctgct actactgcaa cggccccatc
3841 ctggataaag tggtgacagc ccttgaccgg acgtggcacc ctgaacactt cttctgtgca
3901 cagtgtggag ccttctttgg tcccgaaggg ttccacgaga aggacggcaa ggcctactgt
3961 cgcaaggact acttcgacat gttcgcaccc aagtgtggcg gctgcgcccg ggccatcctg
4021 gagaactata tctcagccct caacacgctg tggcatcctg agtgctttgt gtgccgggaa
4081 tgcttcacgc cattcgtgaa cggcagcttc ttcgagcacg acgggcagcc ctactgtgag
4141 gtgcactacc acgagcggcg cggctcgctg tgttctggct gccagaagcc catcaccggc
4201 cgctgcatca ccgccatggc caagaagttc caccccgagc acttcgtctg tgccttctgc
4261 ctcaagcagc tcaacaaggg caccttcaag gagcagaacg acaagcctta ctgtcagaac
4321 tgcttcctca agctcttctg ctaggctcga gtctagagtc gacctgcaga agctagcttg
4381 gccgccatgg cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
4441 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
4501 catcaatgta tcttatcatg tctggatcgg gaattaattc ggcgcagcac catggcctga
4561 aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg
4621 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
4681 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
4741 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
4801 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
4861 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
4921 aggaggcttt tttggaggcc taggcttttg caaaaagctg ttaacagctt ggcactggcc
4981 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
5041 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
5101 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
5161 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
5221 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
5281 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
5341 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
5401 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
5461 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
5521 gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
5581 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
5641 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
5701 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
5761 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
5821 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
5881 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
5941 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
6001 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
6061 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
6121 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
6181 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
6241 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
6301 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
6361 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
6421 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
6481 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
6541 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
6601 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
6661 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
6721 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
6781 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
6841 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
6901 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
6961 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
7021 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
