Download PichiaPink Expression System

Cloning into pPinkα-HC, continued
MCS of pPinkα-HC Below is the multiple cloning site (MCS) for pPinkα-HC. Restriction sites are labeled
to indicate the cleavage site. The multiple cloning site has been confirmed by
sequencing and functional testing. The vector sequence of pPinkα-HC is available
for downloading at www.invitrogen.com or from Technical Support (see page 73).
For a map of pPinkα-HC, see page 58.
5´ end of AOX1 mRNA
5´ AOX1 priming site
811 AACCTTTTTT TTTATCATCA TTATTAGCTT ACTTTCATAA TTGCGACTGG TTCCAATTGA
871 CAAGCTTTTG ATTTTAACGA CTTTTAACGA CAACTTGAGA AGATCAAAAA ACAACTAATT
931 ATTCGAAACG ATG AGA TTT CCT TCA ATT TTT ACT GCT GTT TTA TTC GCA
Met Arg Phe Pro Ser Ile Phe Thr Ala Val Leu Phe Ala
980 GCA TCC TCC GCA TTA GCT GCT CCA GTC AAC ACT ACA ACA GAA GAT GAA
Ala Ser Ser Ala Leu Ala Ala Pro Val Asn Thr Thr Thr Glu Asp Glu
a-factor signal sequence
1028 ACG GCA CAA ATT CCG GCT GAA GCT GTC ATC GGT TAC TCA GAT TTA GAA
Thr Ala Gln Ile Pro Ala Glu Ala Val Ile Gly Tyr Ser Asp Leu Glu
1076 GGG GAT TTC GAT GTT GCT GTT TTG CCA TTT TCC AAC AGC ACA AAT AAC
Gly Asp Phe Asp Val Ala Val Leu Pro Phe Ser Asn Ser Thr Asn Asn
a-factor priming site
1124 GGG TTA TTG TTT ATA AAT ACT ACT ATT GCC AGC ATT GCT GCT AAA GAA
Gly Leu Leu Phe Ile Asn Thr Thr Ile Ala Ser Ile Ala Ala Lys Glu
Stu I
Kpn I
Nae I
Swa I
1172 GAA GGG GTA TCT CTC GAG AAA AGG CCTCAGGTAC CGGCCGGCCATT TAAATACAGG
Glu Gly Val Ser Leu Glu Lys Arg
CYC1 transcription termination region
1228 CCCCTTTTCC TTTGTCGATA TCATGTAATT AGTTATGTCA CGCTTACATT CACGCCCTCC
Continued on next page
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