Part:BBa_K3730000:Design

sPrcn+T7+RBS

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

The introduction of T7 promoter and the shortening of Prcn were with the help of previous research.[1]

Source

We first obtained the plasmid backbone of pSB1C3 via PCR, using primers(5’-3’) below: RBS_EGFP_F: agaaagaggagaaatactagATGAGCAAGGGC BioBrick_prefix_R: CTCTAGAAGCGGCCGCGAATTC

And we obtained the part BBa_K540001 provided by iGEM via PCR, using primers(5’-3’) below: Prefix_K54: CGGCCGCTTCTAGAGttatt RBS_Prcn_R: tctcctctttctgaagaagagttgt

And then through recombination we constructed the plasmid Prcn_eGFP_pSB1C3.

We derived the sequence of T7 promoter from the part BBa_I712074 and we designed that directly into our primers.

On the basis of Prcn_eGFP_pSB1C3, we then constructed T7_sPrcn_eGFP_pSB1C3 via a simple PCR experiment using primers(5’-3’) below: recombine_T7-PSB1C3_F: taatacgactcactataggtactggggggtagtatcaggtact recombine_T7-PSB1C3_R: cctatagtgagtcgtattaCTCTAGAAGCGGCCGCGAATTC

References

[1]Han L. (2020). Mining and engineering of biological parts and their application in gene expression regulation system (Jiangnan University) [韩来闯.(2020).生物元件的挖掘、改造及在基因表达调控系统中的应用(博士学位论文,江南大学)] https://kns.cnki.net/KCMS/detail/detail.aspx?dbname=CDFDLAST2021&filename=1020346676.nh