| 质粒名称: | Puro 2.0 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 24970 |
| 存储实验室: | Tyler Jacks |
|
载体骨架基本信息 |
|
| 载体名称: | Puro 2.0 |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 7705 bp |
| 载体类型: | 哺乳细胞表达，Lentiviral, RNAi |
| 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: | 未知 |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | HpaI(未破坏) |
| 3' 克隆位点: | XhoI(未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | MSCV-rev |
|
文献引用方法 |
|
| 材料和方法部分: | Puro 2.0 was a gift from Tyler Jacks (爱迪基因 plasmid # 24970) |
| 参考文献部分: | Tissue-specific p19Arf regulation dictates the response to oncogenic K-ras. Young NP, Jacks T. Proc Natl Acad Sci U S A. 2010 Jun 1. 107(22):10184-9. 10.1073/pnas.1004796107 PubMed 20479239 |
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 Puro 2.0 7705 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Puro 2.0
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7705)
AUTHORS Young NP, Jacks T
TITLE Tissue-specific p19Arf regulation dictates the response to oncogenic
K-ras.
JOURNAL Proc Natl Acad Sci U S A. 2010 Jun 1. 107(22):10184-9.
PUBMED 20479239
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|Puro 2.0|
FEATURES Location/Qualifiers
source 1..7705
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(47..66)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 238..617
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 619..817
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 772..792
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 835..1015
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1062..1187
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1686..1919
/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 2446..2563
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
primer_bind 2853..2873
/label=mU6-F
/note="Mouse U6 promoter, forward primer"
primer_bind 2951..2967
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
promoter 3030..3529
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(3065..3088)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 3462..3481
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
CDS 3573..4172
/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(3573..3592)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4069..4089
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
primer_bind 4503..4519
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 4594..5182
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4647..4667)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5065..5076)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(5185..5201)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
LTR 5711..5891
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
rep_origin complement(5953..6541)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6033..6052)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6712..7572)
/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 7335..7354
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7573..7677)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
1501 aaagtaagac caccgcacag caagcggccg gccgcgctga tcttcagacc tggaggagga
1561 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
1621 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
1681 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
1741 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
1801 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
1861 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
1921 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
1981 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
2041 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
2101 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
2161 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
2221 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
2281 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
2341 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
2401 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
2461 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
2521 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
2581 cagcagagat ccagtttggt tagtaccggg cccgctctag agatccgacg cgccatctct
2641 aggcccgcgc cggccccctc gcacagactt gtgggagaag ctcggctact cccctgcccc
2701 ggttaatttg catataatat ttcctagtaa ctatagaggc ttaatgtgcg ataaaagaca
2761 gataatctgt tctttttaat actagctaca ttttacatga taggcttgga tttctataag
2821 agatacaaat actaaattat tattttaaaa aacagcacaa aaggaaactc accctaactg
2881 taaagtaatt gtgtgttttg agactataaa tatcccttgg agaaaagcct tgttaacgcg
2941 cggtgaccct cgaggtcgac ggtatcgata agctcgcttc acgagattcc agcagcctag
3001 gaagaattcg tcgacctcga aattctaccg ggtaggggag gcgcttttcc caaggcagtc
3061 tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc
3121 tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt
3181 cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc
3241 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac
3301 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt
3361 cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct
3421 caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa
3481 gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcatcca
3541 tctagatctc gatcgagcag ctgaagctta ccatgaccga gtacaagccc acggtgcgcc
3601 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
3661 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
3721 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
3781 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
3841 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
3901 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
3961 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
4021 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
4081 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
4141 cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga
4201 ccgaaaggag cgcacgaccc catgcatcga tgatatcata atttaaacaa gcaaaaccaa
4261 attaagggcc agctcattcc tcccactcat gatctataga tctatagatc tctcgtggga
4321 tcattgtttt tctcttgatt cccactttgt ggttctaagt actgtggttt ccaaatgtgt
4381 cagtttcata gcctgaagaa cgagatcagc agcctctgtt ccacatacac ttcattctca
4441 gtattgtttt gccaagttct aattccatca gaagctggtc gactctagct agatgcgcgg
4501 cccgaggtcg acggtatcga taagctcgct tcacgagatc atgtttaagg gttccggttc
4561 cactaggtac aattcgatat caagcttatc gataatcaac ctctggatta caaaatttgt
4621 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
4681 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
4741 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
4801 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
4861 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
4921 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
4981 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
5041 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
5101 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
5161 tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gatcgagacc tagaaaaaca
5221 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc
5281 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac
5341 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
5401 aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta
5461 cttccctgat tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg
5521 atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga
5581 gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt
5641 attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc
5701 ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
5761 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
5821 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
5881 aatctctagc agcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
5941 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
6001 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
6061 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
6121 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
6181 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
6241 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
6301 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
6361 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
6421 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
6481 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
6541 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
6601 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
6661 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
6721 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
6781 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
6841 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
6901 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
6961 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
7021 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
7081 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
7141 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
7201 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
7261 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
7321 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
7381 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
7441 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
7501 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
7561 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
7621 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
7681 atttccccga aaagtgccac ctgac
//
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