Part:BBa_K2322005

-AOX1 promoter-RBS-gltB-

This circuit contains a AOX1 promoter, a ribosome binding site, a gltB gene, and a terminator. This circuit is used to test the expression of gltB. If the circuit work, the gltB will express normally in GS115.

Name: BBa_K2322005

AOX1 promoter:

Sequence of the primer: ATGATTTCTGGAATTCGCGGCCGCTTCTAGAGATCTAACATCCAAAGACGAAAGGT

Function: AOX1 is a strong promoter in the pichia pastoris. It is highly effective. It can be restricted by glucose, glycerinum, ethyl alcohol. It can be induced by the methanol. In our experiment, our circuit is in an environment containing methanol. In this circuit, it is used to control and increase the expression of gltB, our targeted gene.

RBS (Ribosome Binding Site): A ribosome binding site is a sequence of mRNA. It is used to make sure the ribosome is on the correct position of the mRNA at the beginning of the translation.

Sequence: TCACACAGGAAACA

gltB

Sequence:

AACACACCTTATGACAGTCAGGAATTGACTGTTTCTCTAACGACTTCCCTTTTAGCCTTAAAGATAAAATCCA TTTTAATTTCAGTCATTTAATAAAGAATTTTGCGCTAAAGCACATTTCTGTACCAATAAGCTTGCCATTTGAC CTGTATCAGCTTTCCCGATAAGTTGGAAATCCGCTGGAAGCTTTCTGGATGAGCAGCCTGCTCATCATATTTA TGCAGTAATTGAGATCCCCTCTTCACCGTATTAACCGATGCGAAAAGGACAACAAGGGGGCGAATCGGAGGCG CGCGTATGACACGCAAACCCCGTCGCCACGCTCTTTCTGTGCCCGTGCGCAGCGGTTCGGAAGTGGGGTTCCC GCAGAGCCTGGGGGAGGTTCACGATATGTTGTACGATAAATCCCTTGAGAGGGATAACTGTGGTTTCGGCCTG ATCGCCCACATAGAAGGCGAACCTAGCCACAAGGTAGTGCGTACTGCAATACACGCACTGGCCCGCATGCAGC ACCGTGGCGCGATTCTCGCCGATGGTAAAACCGGCGACGGTTGCGGCTTGCTGTTACAAAAACCGGATCGCTT TTTTCGCATCGTTGCGCAGGAGCGCGGCTGGCGTTTAGCAAAAAACTACGCTGTCGGGATGCTCTTCCTGAAT AAAGATCCTGAACTCGCCGCTGCCGCACGCCGCATCGTTGAAGAAGAACTGCAACGCGAAACCTTGTCGATTG TGGGCTGGCGTGATGTCCCCACTAACGAAGGCGTGCTGGGTGAAATCGCCCTCTCCTCTCTGCCACGCATTGA GCAAATTTTTGTGAACGCCCCGGCAGGCTGGCGTCCACGCGATATGGAGCGCCGTCTGTTTATCGCCCGCCGC CGCATTGAAAAGCGTCTCGAAGCCGACAAAGACTTCTACGTCTGTAGCCTGTCGAATCTGGTGAACATCTATA AAGGTCTGTGTATGCCGACGGATCTGCCGCGCTTTTATCTGGATCTTGCGGACCTGCGTCTGGAATCGGCCAT TTGCCTGTTCCACCAGCGCTTCTCCACTAACACCGTACCGCGCTGGCCGTGGCGCAACCGTTCCGCTATCTGG CGCATAACGGTGAAATCAACACCATCACCGGTAACCGCCAATGGGCGTGCGCGTACTTATAAATTCCAGACAC CGCTTATCCCTGACCTGCACGACGCCGCACCGTTCGTC

Terminator:

Sequence of the primer: TGACACCTTGCCCTTTTTTGCCGGACTGCAGTTAAGAAGAAGAGTTCATAGACAAAAC

Function: The terminator is used to control the expression of our targeted gene, gltB.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1
  Illegal BamHI site found at 938
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1480