pLenti X1 Puro empty (w610-1)

pLenti X1 Puro empty (w610-1)

载体基本信息

质粒名称:	pLenti X1 Puro empty (w610-1)
出品公司:	Ybscience
存储实验室:	Eric Campeau
用途:	第三代慢病毒空控制向量
载体骨架基本信息
载体名称:	p156RRL-sinPPT-CMV-GFP-PRE/Nhe I
载体抗性:	氨苄霉素
空载体大小:	7424 bp
载体类型:	哺乳细胞表达,慢病毒，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:	None
克隆方法:	限制性内切酶
5' 克隆位点:	Sal I(未破坏)
3' 克隆位点:	Xba I (未破坏)
5' 测序引物	n/a
文献引用方法
材料和方法部分:	pLenti X1 Puro empty (w610-1) was a gift from Eric Campeau (Addgene plasmid # 20953)
参考文献部分:	A Versatile Viral System for Expression and Depletion of Proteins in Mammalian Cells. Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, Campisi J, Yaswen P, Cooper PK, Kaufman PD.. PLoS One. 2009 Aug 6;4(8):e6529. 10.1371/journal.pone.0006529 PubMed 19657394

 

载体质粒图谱和多克隆位点信息

载体简介

 

载体序列

LOCUS       pLenti X1 Puro empty (w610-1)	7424 bp 	DNA	SYN
DEFINITION  pLenti X1 Puro empty (w610-1)
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
FEATURES             Location/Qualifiers
     source          1..7424
                     /organism="pLenti X1 Puro empty (w610-1)"
                     /mol_type="other DNA"
     promoter        62..90
                     /label="AmpR_promoter"
     gene            132..992
                     /label="Ampicillin"
                     /gene="Ampicillin"
     CDS             132..992
                     /label="ORF frame 3"
     rep_origin      1147..1766
                     /label="pBR322_origin"
     promoter        2075..2104
                     /label="lac_promoter"
     misc_feature    2118..2140
                     /label="M13_pUC_rev_primer"
     promoter        2139..2157
                     /label="M13_reverse_primer"
     promoter        2174..2193
                     /label="T3_promoter"
     misc_feature    2448..2628
                     /label="HIV-1_5_LTR"
     misc_feature    2448..2628
                     /label="truncHIV-1_3_LTR"
     misc_feature    2736..2780
                     /label="HIV-1_psi_pack"
     CDS             3168..3668
                     /label="ORF frame 3"
     misc_feature    3290..3523
                     /label="RRE"
     CDS             3629..4189
                     /label="ORF frame 2"
     misc_feature    4016..4031
                     /label="cPPT"
     misc_feature    4161..4185
                     /label="attB1"
     misc_feature    4251..4275
                     /label="attB2"
     misc_feature    4308..4883
                     /label="WPRE"
     CDS             4396..5025
                     /label="ORF frame 1"
     misc_feature    complement(4943..4966)
                     /label="MSCV_rev_primer"
     CDS             5188..6027
                     /label="ORF frame 1"
     misc_feature    5340..5359
                     /label="mPGK_F_primer"
     CDS             complement(5353..6078)
                     /label="ORF frame 3"
     gene            5428..6027
                     /label="puro (variant)"
                     /gene="puro (variant)"
     misc_feature    6148..6163
                     /label="cPPT"
     misc_feature    6148..6169
                     /label="U3PPT"
     misc_feature    6218..6398
                     /label="HIV-1_5_LTR"
     misc_feature    6218..6398
                     /label="truncHIV-1_3_LTR"
     terminator      6473..6592
                     /label="SV40_PA_terminator"
     misc_feature    6561..6580
                     /label="EBV_rev_primer"
     rep_origin      6610..6687
                     /label="SV40_origin"
     misc_feature    6672..6691
                     /label="SV40pro_F_primer"
     promoter        complement(6778..6796)
                     /label="T7_promoter"
     promoter        complement(6806..6822)
                     /label="M13_forward20_primer"
     misc_feature    6815..6958
                     /label="lacZ_a"
     misc_feature    complement(6815..6837)
                     /label="M13_pUC_fwd_primer"
     rep_origin      6984..7290
                     /label="f1_origin"
ORIGIN
    1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
   61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
  121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
  181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
  241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
  301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
  361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
  421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
  481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
  541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
  601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
  661 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
  721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
  781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
  841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
  901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
  961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
 1021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
 1081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
 1141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
 1201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
 1261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
 1321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
 1381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
 1441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
 1501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
 1561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
 1621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
 1681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
 1741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
 1801 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
 1861 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
 1921 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
 1981 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
 2041 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
 2101 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
 2161 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat
 2221 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc
 2281 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg
 2341 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc
 2401 gcattgcaga gatattgtat ttaagtgcct agctcgatac aataaacggg tctctctggt
 2461 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
 2521 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
 2581 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
 2641 cagggacctg aaagcgaaag ggaaaccaga gctctctcga cgcaggactc ggcttgctga
 2701 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
 2761 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
 2821 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
 2881 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
 2941 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
 3001 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
 3061 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
 3121 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt
 3181 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
 3241 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
 3301 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcctcaatg acgctgacgg
 3361 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
 3421 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
 3481 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
 3541 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
 3601 tggaacagat tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
 3661 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
 3721 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
 3781 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
 3841 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
 3901 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
 3961 gagagacaga gacagatcca ttcgattagt gaacggatct cgacggttaa cttttaaaag
 4021 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
 4081 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaatttta tTCCAGTGTG
 4141 GTGGAATTCT GCAGATATCA ACAAGTTTGT ACAAAAAAGC AGGCTTTAAA GGAACCAATT
 4201 CAGTCGACTG GATCCGGTAC CGAATTCGCG GCCGCACTCG AGATATCTAG ACCCAGCTTT
 4261 CTTGTACAAA GTGGTTGATA TCCAGCACAG TGGCGGCCGC TCGAcaatca acctctggat
 4321 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
 4381 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
 4441 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
 4501 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
 4561 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
 4621 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
 4681 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
 4741 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
 4801 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
 4861 acgagtcgga tctccctttg ggccgcctcc ccgcctgGAA TTCTACCGGG TAGGGGAGGC
 4921 GCTTTTCCCA AGGCAGTCTG GAGCATGCGC TTTAGCAGCC CCGCTGGGCA CTTGGCGCTA
 4981 CACAAGTGGC CTCTGGCCTC GCACACATTC CACATCCACC GGTAGGCGCC AACCGGCTCC
 5041 GTTCTTTGGT GGCCCCTTCG CGCCACCTTC TACTCCTCCC CTAGTCAGGA AGTTCCCCCC
 5101 CGCCCCGCAG CTCGCGTCGT GCAGGACGTG ACAAATGGAA GTAGCACGTC TCACTAGTCT
 5161 CGTGCAGATG GACAGCACCG CTGAGCAATG GAAGCGGGTA GGCCTTTGGG GCAGCGGCCA
 5221 ATAGCAGCTT TGCTCCTTCG CTTTCTGGGC TCAGAGGCTG GGAAGGGGTG GGTCCGGGGG
 5281 CGGGCTCAGG GGCGGGCTCA GGGGCGGGGC GGGCGCCCGA AGGTCCTCCG GAGGCCCGGC
 5341 ATTCTGCACG CTTCAAAAGC GCACGTCTGC CGCGCTGTTC TCCTCTTCCT CATCTCCGGG
 5401 CCTTTCGACC TGCAGCCCAA GCTTACCATG ACCGAGTACA AGCCCACGGT GCGCCTCGCC
 5461 ACCCGCGACG ACGTCCCCAG GGCCGTACGC ACCCTCGCCG CCGCGTTCGC CGACTACCCC
 5521 GCCACGCGCC ACACCGTCGA TCCGGACCGC CACATCGAGC GGGTCACCGA GCTGCAAGAA
 5581 CTCTTCCTCA CGCGCGTCGG GCTCGACATC GGCAAGGTGT GGGTCGCGGA CGACGGCGCC
 5641 GCGGTGGCGG TCTGGACCAC GCCGGAGAGC GTCGAAGCGG GGGCGGTGTT CGCCGAGATC
 5701 GGCCCGCGCA TGGCCGAGTT GAGCGGTTCC CGGCTGGCCG CGCAGCAACA GATGGAAGGC
 5761 CTCCTGGCGC CGCACCGGCC CAAGGAGCCC GCGTGGTTCC TGGCCACCGT CGGCGTCTCG
 5821 CCCGACCACC AGGGCAAGGG TCTGGGCAGC GCCGTCGTGC TCCCCGGAGT GGAGGCGGCC
 5881 GAGCGCGCCG GGGTGCCCGC CTTCCTGGAG ACCTCCGCGC CCCGCAACCT CCCCTTCTAC
 5941 GAGCGGCTCG GCTTCACCGT CACCGCCGAC GTCGAGGTGC CCGAAGGACC GCGCACCTGG
 6001 TGCATGACCC GCAAGCCCGG TGCCTGACGC CCGCCCCACG ACCCGCAGCG CCCGACCGAA
 6061 AGGAGCGCAC GACCCCATGC ATCTCGAGGG CCCGGTACct ttaagaccaa tgacttacaa
 6121 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctagctca
 6181 ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
 6241 ctgagcctgg gagctctctg gctgcctagg gaacccactg cttaagcctc aataaagctt
 6301 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
 6361 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat
 6421 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat
 6481 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
 6541 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
 6601 gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
 6661 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
 6721 tagtgaggag gcttttttgg aggcctaggc ttttgcgtcg agacgtaccc aattcgccct
 6781 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
 6841 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
 6901 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
 6961 ggcgcgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
 7021 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
 7081 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
 7141 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
 7201 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
 7261 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
 7321 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
 7381 aatttaacgc gaattttaac aaaatattaa cgtttacaat ttcc