Part:BBa_K2889002:Design

pSB1C3-IL7-AS-S1

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 600
  Illegal SpeI site found at 813
  Illegal SpeI site found at 924
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 626
  Illegal SpeI site found at 813
  Illegal SpeI site found at 924
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 600
  Illegal SpeI site found at 813
  Illegal SpeI site found at 924
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 600
  Illegal SpeI site found at 813
  Illegal SpeI site found at 924
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

We wanted to investigate which domain is essential for the function of IL7-AS. We cloned the truncated sequences of IL7-AS (IL7-AS-S1) into pSB1C3 for submitting to IGEM 2018 and inserted IL7-AS-S1 into pCDNA3.1 to study the function.

1.1 Amplification of IL7-AS-S1 fragments. First we amplified IL7-AS-S1 fragment (line 3) using primers. After that, we purified the PCR products by PCR Purification Kit and digested them with EcoRI and PstI (Fig 1).

1.2 Digested pSB1C3 vector. We digested the pSB1C3 vectors with EcoRI and PstI (Fig 2).

1.3 Ligation of purified IL7-AS-S2; IL7-AS-S1 and IL7-AS fragments to PSB1C3 vector. IL7-AS-S2; IL7-AS-S1 and IL7-AS fragments were ligated to PSB1C3 vector, respectively. Then we selected the positive clones by PCR and sequencing (Fig 3).

The secondary structure of IL7-AS-S1(Fig 4).

Source

We cloned the truncated sequences of IL7-AS (IL7-AS-S1, 1099 bp) from human cells