| 质粒名称: | pSIN4-EF1a-LMO2-IRES-Puro |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 61064 |
| 存储实验室: | Igor Slukvin |
|
载体骨架基本信息 |
|
| 载体名称: | pSIN4-EF1alpha-IRES-Puro |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 7500 bp |
| 载体类型: | 慢病毒表达 |
| 筛选标记: | 嘌呤霉素 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | LMO2 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 2303 bp |
| 启动子 | EF1alpha |
| 标签/融合蛋白: | Flag （插入N端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI (未破坏) |
| 3' 克隆位点: | MluI (未破坏) |
| 5' 测序引物 | EF1a-Fwd |
| 3' 测序引物 | pSIN-R: CAAACGCACACCGGCCTTATT |
|
文献引用方法 |
|
| 材料和方法部分: | pSIN4-EF1a-LMO2-IRES-Puro was a gift from Igor Slukvin (Addgene plasmid # 61064 ; http://n2t.net/addgene:61064 ; RRID:Addgene_61064) |
| 参考文献部分: | Direct induction of haematoendothelial programs in human pluripotent stem cells by transcriptional regulators. Elcheva I, Brok-Volchanskaya V, Kumar A, Liu P, Lee JH, Tong L, Vodyanik M, Swanson S, Stewart R, Kyba M, Yakubov E, Cooke J, Thomson JA, Slukvin I. Nat Commun. 2014 Jul 14;5:4372. doi: 10.1038/ncomms5372. 10.1038/ncomms5372 PubMed 25019369 |
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 pSIN4-EF1a-LMO2-IRES-Puro 8044 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION lentiviral vector for LMO2.
ACCESSION .
VERSION .
KEYWORDS pSIN4-EF1a-LMO2-IRES-Puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8044)
AUTHORS Elcheva I, Brok-Volchanskaya V, Kumar A, Liu P, Lee JH, Tong L,
Vodyanik M, Swanson S, Stewart R, Kyba M, Yakubov E, Cooke J,
Thomson JA, Slukvin I
TITLE Direct induction of haematoendothelial programs in human pluripotent
stem cells by transcriptional regulators.
JOURNAL Nat Commun. 2014 Jul 14;5:4372. doi: 10.1038/ncomms5372.
PUBMED 25019369
REFERENCE 2 (bases 1 to 8044)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8044
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 117..750
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 797..922
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1415..1648
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 2164..2281
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2410..3588
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 2641..3579
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 3536..3556
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
regulatory 3599..3608
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3608..3631
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
misc_feature 4160..4733
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(4327..4344)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 4554..4573
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 4766..5365
/codon_start=1
/gene="pac from Streptomyces"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
primer_bind complement(4766..4785)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 5262..5282
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 5568..5801
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(5952..5969)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5958..6182
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
regulatory 6225..6234
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
promoter 6278..6382
/gene="bla"
/label=AmpR promoter
CDS 6383..7243
/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(6601..6620)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7414..8002
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7903..7922
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cggaccgcca ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa
61 ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagttgga
121 agggctaatt cactcccaaa gaagacaaga tatccttgat ctgtggatct accacacaca
181 aggctacttc cctgattagc agaactacac accagggcca ggggtcagat atccactgac
241 ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag aggccaataa
301 aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg acccggagag
361 agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg cccgagagct
421 gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac tttccgctgg
481 ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatcctg
541 catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct
601 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
661 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
721 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
781 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
841 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
901 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
961 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
1021 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
1081 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
1141 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
1201 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
1261 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
1321 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
1381 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
1441 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
1501 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
1561 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
1621 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
1681 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
1741 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
1801 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
1861 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
1921 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
1981 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
2041 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
2101 gatccattcg attagtgaac ggatctcgac ggtatcgcca caaatggcag tattcatcca
2161 caattttaaa agaaaggggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
2221 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
2281 tcgggtttat tacagggaca gcagagatcc actttggatc gataagcttt gcaaagatgg
2341 ataaagtttt aaacagagag gaatctttgc agctaatgga ccttctaggt cttgaaagga
2401 gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
2461 aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac
2521 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
2581 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
2641 gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg
2701 ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga
2761 agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt
2821 gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt
2881 ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt
2941 tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt
3001 tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg
3061 ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct
3121 ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg
3181 tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca
3241 aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg
3301 gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg
3361 cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt
3421 tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac
3481 ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag
3541 cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgagc cggaattcgc
3601 caccatggac tacaaggacg acgatgacaa gtcctcggcc atcgaaagga agagcctgga
3661 cccttcagag gaaccagtgg atgaggtgct gcagatcccc ccatccctgc tgacatgcgg
3721 cggctgccag cagaacattg gggaccgcta cttcctgaag gccatcgacc agtactggca
3781 cgaggactgc ctgagctgcg acctctgtgg ctgccggctg ggtgaggtgg ggcggcgcct
3841 ctactacaaa ctgggccgga agctctgccg gagagactat ctcaggcttt ttgggcaaga
3901 cggtctctgc gcatcctgtg acaagcggat tcgtgcctat gagatgacaa tgcgggtgaa
3961 agacaaagtg tatcacctgg aatgtttcaa atgcgccgcc tgtcagaagc atttctgtgt
4021 aggtgacaga tacctcctca tcaactctga catagtgtgc gaacaggaca tctacgagtg
4081 gactaagatc aatgggatga tatagacgcg tcggaaacta gttcgaagga tccgcatgca
4141 tctagggcgg ccaattccgc ccctctccct cccccccccc taacgttact ggccgaagcc
4201 gcttggaata aggccggtgt gcgtttgtct atatgtgatt ttccaccata ttgccgtctt
4261 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
4321 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
4381 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
4441 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
4501 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
4561 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
4621 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
4681 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taagcttgcc
4741 acaacccaca aggagacgac cttccatgac cgagtacaag cccacggtgc gcctcgccac
4801 ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
4861 cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
4921 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
4981 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
5041 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
5101 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
5161 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
5221 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
5281 gcggctcggc ttcaccgtca ccgccgacgt cgagtgcccg aaggaccgcg cgacctggtg
5341 catgacccgc aagcccggtg cctgataata ggcggccgct cgagacctag aaaaacatgg
5401 agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca
5461 agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta
5521 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
5581 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca
5641 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
5701 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
5761 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
5821 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggccttgaca
5881 ttataataga tttagcagga attgaactag gagtggagca cacaggcaaa gttctagagc
5941 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
6001 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
6061 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
6121 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
6181 ggcttctgag gcggaaagaa ccagctgggg gcgcgcccct cgaggccgcc atggtcatag
6241 ctgtttgacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
6301 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
6361 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
6421 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
6481 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
6541 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
6601 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
6661 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
6721 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
6781 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
6841 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
6901 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
6961 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
7021 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
7081 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
7141 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
7201 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
7261 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
7321 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
7381 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
7441 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
7501 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
7561 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
7621 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
7681 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
7741 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
7801 tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
7861 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
7921 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
7981 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
8041 ttgc
//
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