TOPO TA Cloning (Invitrogen)
TOPO TA Cloning (Invitrogen)
TOPO TA Cloning (Invitrogen)
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Map of pCR ® 2.1-<strong>TOPO</strong> ®pCR ® 2.1-<strong>TOPO</strong> ®MapThe map below shows the features of pCR ® 2.1-<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. Thearrow indicates the start of transcription for T7 polymerase. For the full sequence of thevector, you may download it from our web site or call Technical Service (page 19).205lacZa ATGM13 Reverse PrimerHind IIIKpn I Sac I BamH I Spe ICAG GAA ACA GCT ATG ACC ATG ATT ACG CCA AGC TTG G<strong>TA</strong> CCG AGC TCG GAT CCA C<strong>TA</strong>GTC CTT TGT CGA <strong>TA</strong>C TGG <strong>TA</strong>C <strong>TA</strong>A TGC GGT TCG AAC CAT GGC TCG AGC C<strong>TA</strong> GGT GATBstX I EcoR IEcoR IG<strong>TA</strong> ACG GCC GCC AGT GTG CTG GAA TTC GCC CTT PCR Product AAG GGC GAA TTC TGCCAT TGC CGG CGG TCA CAC GAC CTT AAG CGG GAATTC CCG CTT AAG ACGEcoR VBstX INot I Xho I Nsi I Xba I Apa IAGA <strong>TA</strong>T CCA TCA CAC TGG CGG CCG CTC GAG CAT GCA TCT AGA GGG CCC AAT TCG CCC <strong>TA</strong>TTCT A<strong>TA</strong> GGT 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 Forward (-20) Primer M13 Forward (-40) 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 TTG426+1PlaclacZaf1 oriComments for pCR ® 2.1-<strong>TOPO</strong> ®3908 nucleotidespUC oriAmpicillinLacZa fragment: bases 1-571M13 reverse priming site: bases 205-221Multiple cloning site: bases 234-357T7 promoter/priming site: bases 364-383M13 Forward (-20) priming site: bases 391-406M13 Forward (-40) priming site: bases 411-426f1 origin: bases 548-962Kanamycin resistance ORF: bases 1296-2090Ampicillin resistance ORF: bases 2108-2968pUC origin: bases 3113-3786pCR ® 2.1-<strong>TOPO</strong> ®3.9 kbKanamycin10
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