TOPO TA Cloning (Invitrogen)
TOPO TA Cloning (Invitrogen)
TOPO TA Cloning (Invitrogen)
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Map of pCR ® II-<strong>TOPO</strong> ®pCR ® II-<strong>TOPO</strong> ®MapThe map below shows the features of pCR ® II-<strong>TOPO</strong> ® and the sequence surrounding the<strong>TOPO</strong> ® <strong>Cloning</strong> site. Restriction sites are labeled to indicate the actual cleavage site. Thearrows indicate the start of transcription for Sp6 and T7 polymerases. For the fullsequence of the vector, you may download it from our web site or call Technical Service(page 19).lacZa ATGM13 Reverse PrimerSp6 Promoter205 CAG GAA ACA GCT ATG ACC ATG ATT ACG CCA AGC <strong>TA</strong>T T<strong>TA</strong> GGT GAC ACT A<strong>TA</strong> GAAGTC CTT TGT CGA <strong>TA</strong>C TGG <strong>TA</strong>C <strong>TA</strong>A TGC GGT TCG A<strong>TA</strong> AAT CCA CTG TGA <strong>TA</strong>T CTTNsi I Hind III Kpn I Sac I BamH I Spe I<strong>TA</strong>C TCA AGC <strong>TA</strong>T GCA TCA AGC TTG G<strong>TA</strong> CCG AGC TCG GAT CCA C<strong>TA</strong> G<strong>TA</strong> ACG GCCATG AGT TCG A<strong>TA</strong> CGT AGT TCG AAC CAT GGC TCG AGC C<strong>TA</strong> GGT GAT CAT TGC CGGBstX I EcoR I EcoR I EcoR VGCC AGT GTG CTG GAA TTC GCC CTTCGG TCA CAC GAC CTT AAG CGG GAAPCR ProductAAG GGC GAA TTC TGC AGA <strong>TA</strong>TTTC CCG CTT AAG ACG TCT A<strong>TA</strong>BstX I Not I Xho I Nsi I Xba I Apa ICCA TCA CAC TGG CGG CCG CTC GAG CAT GCA TCT AGA GGG CCC AAT TCG CCC <strong>TA</strong>TGGT AGT GTG ACC GCC GGC GAG CTC G<strong>TA</strong> CGT AGA TCT CCC GGG T<strong>TA</strong> AGC GGG A<strong>TA</strong>T7 Promoter M13 (-20) Forward Primer M13(-40) Forward PrimerAGT GAG TCG <strong>TA</strong>T <strong>TA</strong>C AAT TCA CTG GCC GTC GTT T<strong>TA</strong> CAA CGT CGT GAC TGG GAA AACTCA CTC AGC A<strong>TA</strong> ATG T<strong>TA</strong> AGT GAC CGG CAG CAA AAT GTT GCA GCA CTG ACC CTT TTG468+1PlaclacZf1 oriComments for pCR ® II-<strong>TOPO</strong> ®3950 nucleotidespUC oriLacZa gene: bases 1-588M13 Reverse priming site: bases 205-221Sp6 promoter: bases 239-256Multiple <strong>Cloning</strong> Site: bases 269-399T7 promoter: bases 406-425M13 (-20) Forward priming site: bases 433-448M13 (-40) Forward priming site: bases 453-468f1 origin: bases 590-1004Kanamycin resistance ORF: bases 1338-2132Ampicillin resistance ORF: bases 2150-3010pUC origin: bases 3155-3828pCR ® II-<strong>TOPO</strong> ®3.9 kbAmpicillinKanamycin11
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