lenti sgRNA(MS2)_zeo backbone載體

CRISPR/Cas9系統質粒，表達guideRNA，含有MS2 loop元件，U6啟動子，EF-1a啟動子驅動Zeocin真核抗性基因表達。

PURPOSE

(Empty Backbone) lenti sgRNA cloning backbone with MS2 loops at tetraloop and stemloop 2 and EF1a-zeo resistance marker. Contains BsmBI sites for insertion of spacer sequences.

PUBLICATION

Konermann et al Nature. 2014 Dec 10. doi: 10.1038/nature14136.

BACKBONE

Vector backboneplenti

Vector typeMammalian Expression, Lentiviral, CRISPR

PromoterU6 and EF1A

Selectable markersZeocin

GROWTH IN BACTERIA

Bacterial Resistance(s)Ampicillin

Growth Temperature37°C

Growth Strain(s)Stbl3

Growth instructionsnone

Copy numberHigh Copy

CLONING INFORMATION

Cloning methodRestriction Enzyme

5′ sequencing primerGAG GGC CTA TTT CCC ATG ATT CCT TCA TAT

3′ sequencing primercctagaaggtccattagctgcaaagattcc

Map：

BioVector NTCC保藏中心提供

Sequence

gtcgacggatcgggagatctcccgatcccctatggtgcactctcagtacaatctgctctgatgccgcata

gttaagccagtatctgctccctgcttgtgtgttggaggtcgctgagtagtgcgcgagcaaaatttaagct

acaacaaggcaaggcttgaccgacaattgcatgaagaatctgcttagggttaggcgttttgcgctgcttc

gcgatgtacgggccagatatacgcgttgacattgattattgactagttattaatagtaatcaattacggg

gtcattagttcatagcccatatatggagttccgcgttacataacttacggtaaatggcccgcctggctga

ccgcccaacgacccccgcccattgacgtcaataatgacgtatgttcccatagtaacgccaatagggactt

tccattgacgtcaatgggtggagtatttacggtaaactgcccacttggcagtacatcaagtgtatcatat

gccaagtacgccccctattgacgtcaatgacggtaaatggcccgcctggcattatgcccagtacatgacc

ttatgggactttcctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggtttt

ggcagtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgt

caatgggagtttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattg

acgcaaatgggcggtaggcgtgtacggtgggaggtctatataagcagcgcgttttgcctgtactgggtct

ctctggttagaccagatctgagcctgggagctctctggctaactagggaacccactgcttaagcctcaat

aaagcttgccttgagtgcttcaagtagtgtgtgcccgtctgttgtgtgactctggtaactagagatccct

cagacccttttagtcagtgtggaaaatctctagcagtggcgcccgaacagggacttgaaagcgaaaggga

aaccagaggagctctctcgacgcaggactcggcttgctgaagcgcgcacggcaagaggcgaggggcggcg

actggtgagtacgccaaaaattttgactagcggaggctagaaggagagagatgggtgcgagagcgtcagt

attaagcgggggagaattagatcgcgatgggaaaaaattcggttaaggccagggggaaagaaaaaatata

aattaaaacatatagtatgggcaagcagggagctagaacgattcgcagttaatcctggcctgttagaaac

atcagaaggctgtagacaaatactgggacagctacaaccatcccttcagacaggatcagaagaacttaga

tcattatataatacagtagcaaccctctattgtgtgcatcaaaggatagagataaaagacaccaaggaag

ctttagacaagatagaggaagagcaaaacaaaagtaagaccaccgcacagcaagcggccgctgatcttca

gacctggaggaggagatatgagggacaattggagaagtgaattatataaatataaagtagtaaaaattga

accattaggagtagcacccaccaaggcaaagagaagagtggtgcagagagaaaaaagagcagtgggaata

ggagctttgttccttgggttcttgggagcagcaggaagcactatgggcgcagcgtcaatgacgctgacgg

tacaggccagacaattattgtctggtatagtgcagcagcagaacaatttgctgagggctattgaggcgca

acagcatctgttgcaactcacagtctggggcatcaagcagctccaggcaagaatcctggctgtggaaaga

tacctaaaggatcaacagctcctggggatttggggttgctctggaaaactcatttgcaccactgctgtgc

cttggaatgctagttggagtaataaatctctggaacagatttggaatcacacgacctggatggagtggga

cagagaaattaacaattacacaagcttaatacactccttaattgaagaatcgcaaaaccagcaagaaaag

aatgaacaagaattattggaattagataaatgggcaagtttgtggaattggtttaacataacaaattggc

tgtggtatataaaattattcataatgatagtaggaggcttggtaggtttaagaatagtttttgctgtact

ttctatagtgaatagagttaggcagggatattcaccattatcgtttcagacccacctcccaaccccgagg

ggacccgacaggcccgaaggaatagaagaagaaggtggagagagagacagagacagatccattcgattag

tgaacggatcggcactgcgtgcgccaattctgcagacaaatggcagtattcatccacaattttaaaagaa

aaggggggattggggggtacagtgcaggggaaagaatagtagacataatagcaacagacatacaaactaa

agaattacaaaaacaaattacaaaaattcaaaattttcgggtttattacagggacagcagagatccagtt

tggttaattagctagcgagggcctatttcccatgattccttcatatttgcatatacgatacaaggctgtt

agagagataattggaattaatttgactgtaaacacaaagatattagtacaaaatacgtgacgtagaaagt

aataatttcttgggtagtttgcagttttaaaattatgttttaaaatggactatcatatgcttaccgtaac

ttgaaagtatttcgatttcttggctttatatatcttGTGGAAAGGACGAAACACCgGAGACGggatacCG

TCTCtgttttagagctaggccAACATGAGGATCACCCATGTCTGCAGggcctagcaagttaaaataaggc

tagtccgttatcaacttggccAACATGAGGATCACCCATGTCTGCAGggccaagtggcaccgagtcggtg

cTTTTTTTggatcctgcaaagatggataaagttttaaacagagaggaatctttgcagctaatggaccttc

taggtcttgaaaggagtgggaattggctccggtgcccgtcagtgggcagagcgcacatcgcccacagtcc

ccgagaagttggggggaggggtcggcaattgatccggtgcctagagaaggtggcgcggggtaaactggga

aagtgatgtcgtgtactggctccgcctttttcccgagggtgggggagaaccgtatataagtgcagtagtc

gccgtgaacgttctttttcgcaacgggtttgccgccagaacacaggtaagtgccgtgtgtggttcccgcg

ggcctggcctctttacgggttatggcccttgcgtgccttgaattacttccactggctgcagtacgtgatt

cttgatcccgagcttcgggttggaagtgggtgggagagttcgaggccttgcgcttaaggagccccttcgc

ctcgtgcttgagttgaggcctggcctgggcgctggggccgccgcgtgcgaatctggtggcaccttcgcgc

ctgtctcgctgctttcgataagtctctagccatttaaaatttttgatgacctgctgcgacgctttttttc

tggcaagatagtcttgtaaatgcgggccaagatctgcacactggtatttcggtttttggggccgcgggcg

gcgacggggcccgtgcgtcccagcgcacatgttcggcgaggcggggcctgcgagcgcggccaccgagaat

cggacgggggtagtctcaagctggccggcctgctctggtgcctggcctcgcgccgccgtgtatcgccccg

ccctgggcggcaaggctggcccggtcggcaccagttgcgtgagcggaaagatggccgcttcccggccctg

ctgcagggagctcaaaatggaggacgcggcgctcgggagagcgggcgggtgagtcacccacacaaaggaa

aagggcctttccgtcctcagccgtcgcttcatgtgactccacggagtaccgggcgccgtccaggcacctc

gattagttctcgagcttttggagtacgtcgtctttaggttggggggaggggttttatgcgatggagtttc

cccacactgagtgggtggagactgaagttaggccagcttggcacttgatgtaattctccttggaatttgc

cctttttgagtttggatcttggttcattctcaagcctcagacagtggttcaaagtttttttcttccattt

caggtgtcgtgatgtacaATGGCCAAGTTGACCAGTGCCGTTCCGGTGCTCACCGCGCGCGACGTCGCCG

GAGCGGTCGAGTTCTGGACCGACCGGCTCGGGTTCTCCCGGGACTTCGTGGAGGACGACTTCGCCGGTGT

GGTCCGGGACGACGTGACCCTGTTCATCAGCGCGGTCCAGGACCAGGTGGTGCCGGACAACACCCTGGCC

TGGGTGTGGGTGCGCGGCCTGGACGAGCTGTACGCCGAGTGGTCGGAGGTCGTGTCCACGAACTTCCGGG

ACGCCTCCGGGCCGGCCATGACCGAGATCGGCGAGCAGCCGTGGGGGCGGGAGTTCGCCCTGCGCGACCC

GGCCGGCAACTGCGTGCACTTCGTGGCCGAGGAGCAGGACTGAgaattcgatatcaagcttatcggtaat

caacctctggattacaaaatttgtgaaagattgactggtattcttaactatgttgctccttttacgctat

gtggatacgctgctttaatgcctttgtatcatgctattgcttcccgtatggctttcattttctcctcctt

gtataaatcctggttgctgtctctttatgaggagttgtggcccgttgtcaggcaacgtggcgtggtgtgc

actgtgtttgctgacgcaacccccactggttggggcattgccaccacctgtcagctcctttccgggactt

tcgctttccccctccctattgccacggcggaactcatcgccgcctgccttgcccgctgctggacaggggc

tcggctgttgggcactgacaattccgtggtgttgtcggggaaatcatcgtcctttccttggctgctcgcc

tgtgttgccacctggattctgcgcgggacgtccttctgctacgtcccttcggccctcaatccagcggacc

ttccttcccgcggcctgctgccggctctgcggcctcttccgcgtcttcgccttcgccctcagacgagtcg

gatctccctttgggccgcctccccgcatcgataccgtcgacctcgagacctagaaaaacatggagcaatc

acaagtagcaatacagcagctaccaatgctgattgtgcctggctagaagcacaagaggaggaggaggtgg

gttttccagtcacacctcaggtacctttaagaccaatgacttacaaggcagctgtagatcttagccactt

tttaaaagaaaaggggggactggaagggctaattcactcccaacgaagacaagatatccttgatctgtgg

atctaccacacacaaggctacttccctgattggcagaactacacaccagggccagggatcagatatccac

tgacctttggatggtgctacaagctagtaccagttgagcaagagaaggtagaagaagccaatgaaggaga

gaacacccgcttgttacaccctgtgagcctgcatgggatggatgacccggagagagaagtattagagtgg

aggtttgacagccgcctagcatttcatcacatggcccgagagctgcatccggactgtactgggtctctct

ggttagaccagatctgagcctgggagctctctggctaactagggaacccactgcttaagcctcaataaag

cttgccttgagtgcttcaagtagtgtgtgcccgtctgttgtgtgactctggtaactagagatccctcaga

cccttttagtcagtgtggaaaatctctagcagggcccgtttaaacccgctgatcagcctcgactgtgcct

tctagttgccagccatctgttgtttgcccctcccccgtgccttccttgaccctggaaggtgccactccca

ctgtcctttcctaataaaatgaggaaattgcatcgcattgtctgagtaggtgtcattctattctgggggg

tggggtggggcaggacagcaagggggaggattgggaagacaatagcaggcatgctggggatgcggtgggc

tctatggcttctgaggcggaaagaaccagctggggctctagggggtatccccacgcgccctgtagcggcg

cattaagcgcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgccagcgccctagcgcccgc

tcctttcgctttcttcccttcctttctcgccacgttcgccggctttccccgtcaagctctaaatcggggg

ctccctttagggttccgatttagtgctttacggcacctcgaccccaaaaaacttgattagggtgatggtt

cacgtagtgggccatcgccctgatagacggtttttcgccctttgacgttggagtccacgttctttaatag

tggactcttgttccaaactggaacaacactcaaccctatctcggtctattcttttgatttataagggatt

ttgccgatttcggcctattggttaaaaaatgagctgatttaacaaaaatttaacgcgaattaattctgtg

gaatgtgtgtcagttagggtgtggaaagtccccaggctccccagcaggcagaagtatgcaaagcatgcat

ctcaattagtcagcaaccaggtgtggaaagtccccaggctccccagcaggcagaagtatgcaaagcatgc

atctcaattagtcagcaaccatagtcccgcccctaactccgcccatcccgcccctaactccgcccagttc

cgcccattctccgccccatggctgactaattttttttatttatgcagaggccgaggccgcctctgcctct

gagctattccagaagtagtgaggaggcttttttggaggcctaggcttttgcaaaaagctcccgggagctt

gtatatccattttcggatctgatcagcacgtgttgacaattaatcatcggcatagtatatcggcatagta

taatacgacaaggtgaggaactaaaccatggccaagttgaccagtgccgttccggtgctcaccgcgcgcg

acgtcgccggagcggtcgagttctggaccgaccggctcgggttctcccgggacttcgtggaggacgactt

cgccggtgtggtccgggacgacgtgaccctgttcatcagcgcggtccaggaccaggtggtgccggacaac

accctggcctgggtgtgggtgcgcggcctggacgagctgtacgccgagtggtcggaggtcgtgtccacga

acttccgggacgcctccgggccggccatgaccgagatcggcgagcagccgtgggggcgggagttcgccct

gcgcgacccggccggcaactgcgtgcacttcgtggccgaggagcaggactgacacgtgctacgagatttc

gattccaccgccgccttctatgaaaggttgggcttcggaatcgttttccgggacgccggctggatgatcc

tccagcgcggggatctcatgctggagttcttcgcccaccccaacttgtttattgcagcttataatggtta

caaataaagcaatagcatcacaaatttcacaaataaagcatttttttcactgcattctagttgtggtttg

tccaaactcatcaatgtatcttatcatgtctgtataccgtcgacctctagctagagcttggcgtaatcat

ggtcatagctgtttcctgtgtgaaattgttatccgctcacaattccacacaacatacgagccggaagcat

aaagtgtaaagcctggggtgcctaatgagtgagctaactcacattaattgcgttgcgctcactgcccgct

ttccagtcgggaaacctgtcgtgccagctgcattaatgaatcggccaacgcgcggggagaggcggtttgc

gtattgggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggt

atcagctcactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgtga

gcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcc

cccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagata

ccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctg

tccgcctttctcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgt

aggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccgg

taactatcgtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacagg

attagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacacta

gaagaacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttg

atccggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaa

aaaggatctcaagaagatcctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgtt

aagggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttt

taaatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaatcagtgaggcacct

atctcagcgatctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataactacgatac

gggagggcttaccatctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccagattt

atcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaactttatccgcctccatc

cagtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgcaacgttgttg

ccattgctacaggcatcgtggtgtcacgctcgtcgtttggtatggcttcattcagctccggttcccaacg

atcaaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctccttcggtcctccgatcgtt

gtcagaagtaagttggccgcagtgttatcactcatggttatggcagcactgcataattctcttactgtca

tgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcg

gcgaccgagttgctcttgcccggcgtcaatacgggataataccgcgccacatagcagaactttaaaagtg

ctcatcattggaaaacgttcttcggggcgaaaactctcaaggatcttaccgctgttgagatccagttcga

tgtaacccactcgtgcacccaactgatcttcagcatcttttactttcaccagcgtttctgggtgagcaaa

aacaggaaggcaaaatgccgcaaaaaagggaataagggcgacacggaaatgttgaatactcatactcttc

ctttttcaatattattgaagcatttatcagggttattgtctcatgagcggatacatatttgaatgtattt

agaaaaataaacaaataggggttccgcgcacatttccccgaaaagtgccacctgac