Part:BBa_K2322002

-AOX1 promoter-RBS-ScTPS1-

This circuit contains a AOX1 promoter, a ribosome binding site, a ScTPS1 gene, and a terminator. This circuit is used to test the expression of ScTPS1. If the circuit work, the ScTPS1 will express normally in GS115 and affect the amount of trehalose in GS115.

Name: BBa_K2322002

AOX1 promoter: Sequence of the primer: ATGATTTCTGGAATTCGCGGCCGCTTCTAGAATGACTACAGATAATGCTAAAGCAC

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 ScTPS1, 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

ScTPS1: ATGACTACGGATAACGCTAAGGCGCAACTGACCTCGTCTTCAGGGGGTAACATTATTGTGGTGTCCAACAGGCTTCCCGTGACAATCACTAAAAACAG CAGTACGGGACAGTACGAGTACGCAATGTCGTCCGGAGGGCTGGTCACGGCGTTGGAAGGGTTGAAGAAGACGTACACTTTCAAGTGGTTCGGATGGC CTGGGCTAGAGATTCCTGACGATGAGAAGGATCAGGTGAGGAAGGACTTGCTGGAAAAGTTTAATGCCGTACCCATCTTCCTGAGCGATGAAATCGCA GACTTACACTACAACGGGTTCAGTAATTCTATTCTATGGCCGTTATTCCATTACCATCCTGGTGAGATCAATTTCGACGAGAATGCGTGGTTGGCATA CAACGAGGCAAACCAGACGTTCACCAACGAGATTGCTAAGACTATGAACCATAACGATTTAATCTGGGTGCATGATTACCATTTGATGTTGGTTCCGG AAATGTTGAGAGTCAAGATTCACGAGAAGCAACTGCAAAACGTTAAGGTCGGGTGGTTCTGCACACACCATTCCCTTCGAGTGAAATTTACAGAATCT TACCTGTCAGACAAGAGATTTTGAAGGGTGTTTTGAGTTGTGATTTAGTCGGGTTCCACACATACGATTATGCAAGACATTTCTTGTCTTCCGTGCAA AGAGTGCTTAACGTGAACACATTGCCTAATGGGGTGGAATACCAGGGCAGATTCGTTAACGTAGGGGCCTTCCTATCGGTATCGACGTGGACAAGTTC ACCGATGGGTTGAAAAAGGAATCCGTACAAAAGAGAATCCAACAATTGAAGGAAACTTTCAAGGGCTGCAAGATCATAGTTGGTGTCGACAGGCTGGA TTACATCAAAGGTGTGCCTCAGAAGTTGCACGCCATGGAAGTGTTTCTGAACGAGCATCCAGAATGGAGGGGCAAGGTTGTTCTGGTACAGGTTGCAG TGCCAAGTCGTGGAGATGTGGAAGAGTACCAATATTTAAGATCTGTGGTCAATGAGTTGGTCGGTAGAATCAACGGTCAGTTCGGTACTGTGGAATTC GTCCCCATCCATTTCATGCACAAGTCTATACCATTTGAAGAGCTGATTTCGTTATATGCTGTGAGCGATGTTTGTTTGGTCTCGTCCACCGTGATGGT ATGAACTTGGTTCCTACGAATATATTGCTTGCCAAGAAGAAAAGAAAGGTTCCTTAATCCTGAGTGAGTTCACAGGTGCCGCACAATCCTTGAATGGT GCTATTATTGTAAATCCTTGGAACACCGATGATCTTTCTGATGCCATCAACGAGGCCTTGACTTTGCCCGATGTAAAGAAAGAAGTTAACTGGGAAAA ACTTTACAAATACTCTCTAAATACACTTCTGCCTTCTGGGGTGAAAATTTCGTCCATGAATTATACAGTACATCATCAAGCTCAACAAGCTCCTCTGC CACCAAAAACTGA

Terminator:

Sequence of the primer: TGACACCTTGCCCTTTTTTGCCGGACTGCAGTTAATTTTTAGTTGCTGAAGATGATGATG

Function: The terminator is used to control the expression of our targeted gene, ScTPS1.

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
  COMPATIBLE WITH RFC[1000]