| 质粒名称: | pCAGGS-flpE-puro |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 20733 |
| 存储实验室: | Rudolf Jaenisch |
|
载体骨架基本信息 |
|
| 载体名称: | pBS |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 3176 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 嘌呤霉素 |
| 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: | FlpE recombinase |
| 物种来源: | S. cerevisiae (budding yeast) |
| 插入大小: | 1416 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRV(未破坏) |
| 3' 克隆位点: | SphI(未破坏) |
| 5' 测序引物 | TTCGGCTTCTGGCGTGTGAC |
| 3' 测序引物 | M13R |
|
文献引用方法 |
|
| 材料和方法部分: | pCAGGS-flpE-puro was a gift from Rudolf Jaenisch (Addgene plasmid # 20733 ; http://n2t.net/addgene:20733 ; RRID:Addgene_20733) |
| 参考文献部分: | Efficient method to generate single-copy transgenic mice by site-specific integration in embryonic stem cells. Beard C, Hochedlinger K, Plath K, Wutz A, Jaenisch R. Genesis. 2006 Jan . 44(1):23-8. 10.1002/gene.20180 PubMed 16400644 |
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 pCAGGS-flpE-puro 7728 bp ds-DNA circular SYN 23-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCAGGS-flpE-puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7728)
AUTHORS Beard C, Hochedlinger K, Plath K, Wutz A, Jaenisch R
TITLE Efficient method to generate single-copy transgenic mice by
site-specific integration in embryonic stem cells.
JOURNAL Genesis. 2006 Jan . 44(1):23-8.
PUBMED 16400644
REFERENCE 2 (bases 1 to 7728)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 23, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7728
/organism="synthetic DNA construct"
/mol_type="other DNA"
polyA_signal 514..648
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
enhancer 662..1041
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
intron 1319..2335
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 2259..2281
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 2343..2362
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 2396..2405
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2402..3700
/codon_start=1
/product="nuclear-targeted site-specific recombinase"
/label=FLPe
/note="FLPe is a fusion between the SV40 nuclear
localization signal and a thermostable version of the
Saccharomyces cerevisiae site-specific recombinase FLP.
Recombination occurs at FRT sequences."
/translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
misc_feature 3739..4325
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(3906..3923)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 4133..4152
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 4326..4925
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(4326..4345)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4822..4842
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
primer_bind complement(5006..5023)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5012..5236
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(5283..5299)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5283..5299)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5296..5318)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5307..5323
/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(5331..5361)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5376..5397
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5456..5651
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 5502..5637
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5564..5583
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 5657..5791
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5859..5876)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6030..6618)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6110..6129)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6789..7649)
/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 7412..7431
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
ORIGIN
1 tattcagcta tattcacata gtggtgaggc tgaaatgagg cgggaagagg cggttggggc
61 ttaattatat caatttgggt ggccccacag cgcctccaag gcgccagtcc tgttttgaca
121 agttgcctct ggaagcctct cttctttttc tccagagtaa gcggaggcca ggggcccccg
181 gcctctgctt aatactaaaa aaaacagctg ttgtcatagt aatgattggg tggaaacatt
241 ccaggcctgg gtggagaggc tttttgcttc ctcttgcaaa accacactga cattccaggc
301 ctgggtggag aggctttttg cttcctcttg caaaaccaca ctgccctctg gagggcagtt
361 gcctagcaac taattaaaag aggatgtcgc acggccagct gcggtcagtt agtcacttcc
421 tgcttaactg acttgacatt ttctatttta agagtcggga ggaaaattac tgtgttggag
481 gccctccgcc atcttctgaa gctgaatcga attaacttgt ttattgcagc ttataatggt
541 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
601 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg atatcatcgt
661 cgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
721 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
781 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
841 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
901 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
961 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
1021 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
1081 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
1141 atgggggcgg gggggggggg ggcgcgcgcc aggggggggg gggggggggg gggggggggg
1201 gggggggggg gggggggggg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
1261 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
1321 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
1381 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
1441 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
1501 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt
1561 gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg
1621 cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg
1681 cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg
1741 cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc
1801 ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg
1861 cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg
1921 gccgcctcgg gccggggggg gctcgggggg ggggcgcggc ggcccccgga gcgccggcgg
1981 ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca
2041 gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc
2101 ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
2161 ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg
2221 gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac
2281 cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct
2341 gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattac ccgccgccac
2401 catggctccc aagaagaaga ggaaggtgat gagtcaattt gatatattat gtaaaacacc
2461 acctaaggtc ctggttcgtc agtttgtgga aaggtttgaa agaccttcag gggaaaaaat
2521 agcatcatgt gctgctgaac taacctattt atgttggatg attactcata acggaacagc
2581 aatcaagaga gccacattca tgagctataa tactatcata agcaattcgc tgagtttcga
2641 tattgtcaac aaatcactcc agtttaaata caagacgcaa aaagcaacaa ttctggaagc
2701 ctcattaaag aaattaattc ctgcttggga atttacaatt attccttaca atggacaaaa
2761 acatcaatct gatatcactg atattgtaag tagtttgcaa ttacagttcg aatcatcgga
2821 agaagcagat aagggaaata gccacagtaa aaaaatgctt aaagcacttc taagtgaggg
2881 tgaaagcatc tgggagatca ctgagaaaat actaaattcg tttgagtata cctcgagatt
2941 tacaaaaaca aaaactttat accaattcct cttcctagct actttcatca attgtggaag
3001 attcagcgat attaagaacg ttgatccgaa atcatttaaa ttagtccaaa ataagtatct
3061 gggagtaata atccagtgtt tagtgacaga gacaaagaca agcgttagta ggcacatata
3121 cttctttagc gcaaggggta ggatcgatcc acttgtatat ttggatgaat ttttgaggaa
3181 ttctgaacca gtcctaaaac gagtaaatag gaccggcaat tcttcaagca acaaacagga
3241 ataccaatta ttaaaagata acttagtcag atcgtacaac aaggctttga agaaaaatgc
3301 gccttatcca atctttgcta taaagaatgg cccaaaatct cacattggaa gacatttgat
3361 gacctcattt ctgtcaatga agggcctaac ggagttgact aatgttgtgg gaaattggag
3421 cgataagcgt gcttctgccg tggccaggac aacgtatact catcagataa cagcaatacc
3481 tgatcactac ttcgcactag tttctcggta ctatgcatat gatccaatat caaaggaaat
3541 gatagcattg aaggatgaga ctaatccaat tgaggagtgg cagcatatag aacagctaaa
3601 gggtagtgct gaaggaagca tacgataccc cgcatggaat gggataatat cacaggaggt
3661 actagactac ctttcatcct acataaatag acgcatataa tgatctagag tcgagttaat
3721 taagaattcc gccccccccc cctctccctc ccccccccct aacgttactg gccgaagccg
3781 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
3841 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
3901 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
3961 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
4021 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
4081 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
4141 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
4201 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
4261 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca
4321 caaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
4381 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
4441 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
4501 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
4561 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
4621 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
4681 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
4741 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
4801 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
4861 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
4921 cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat
4981 cgatgatcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
5041 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
5101 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
5161 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
5221 gcggtgggct ctatggcttc tgaggcggaa agaacctgca gcccaagctt ggcgtaatca
5281 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
5341 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
5401 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc
5461 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
5521 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
5581 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
5641 gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag
5701 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
5761 actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg
5821 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
5881 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
5941 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
6001 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
6061 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
6121 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
6181 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
6241 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
6301 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
6361 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
6421 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
6481 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
6541 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
6601 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
6661 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
6721 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
6781 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
6841 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
6901 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
6961 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
7021 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
7081 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
7141 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
7201 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
7261 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
7321 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
7381 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
7441 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
7501 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
7561 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
7621 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatgacc gctttggaaa
7681 aacaaagact gtatttcctg gaaattaatg tttattcaat aaactgtg
//
