Team:BostonU/Data

From 2014.igem.org

• TEAM
• PROJECT
  • Overview
  • Priority Encoder
  • MoClo Assembly Method
  • Low Copy Backbones
  • Tandem Promoters
  • Fusion Proteins
  • Repressor Proteins
  • Multiplexing
  • BioDesign Automation Tools
  • Future Work
• ACHIEVEMENTS
  • Data Collected
  • Parts Submitted
  • Chimera Workflow
  • Chimera Example
  • Medal Fulfillment
• NOTEBOOK
  • Training
  • Protocols
  • Backbones
  • Tandem Promoters and Repressors
  • Fusion Proteins
• CONSIDERATIONS
  • Collaborations
  • Measurement Track
  • Safety
  • Interlab Study
  • NEGEM
  • Policy and Practices
• ACKNOWLEDGEMENTS

Data Collected

Primer Design for Tandem Promoters and Repressor Genes

Device Name	Forward Primer	Sequence	Reverse Primer	Sequence
BetI_CD	BetI_For_C	ATGAAGACGTAATGGTGCCGAAACTGGGTATGCAGAGC	BetI_Rev_D	ACGAAGACCTACCTTTAATCGGTCGGCAGATGCTGGGT
PhlF_CD	PhlF_For_C	ATGAAGACGTAATGATGGCACGTACCCCGAGCCGTAGC	PhlF_Rev_D	ACGAAGACCTACCTTTAACGCTGTGTACCCGGACAAAC
BM3R1_CD	BM3R1_For_C	ATGAAGACGTAATGATGGAAAGCACCCCGACCAAACAG	BM3R1_Rev_D	ACGAAGACCTACCTTTAGCTCTGACGGCTCAGTGCTGC
LmrA_CD	LmrA_For_C	ATGAAGACGTAATGATGAGCTATGGTGATAGCCGTGAA	LmrA_Rev_D	ACGAAGACCTACCTTTAACGTTTCAGCAGATCCGGAAT
SrpR_CD	SrpR_For_C	ATGAAGACGTAATGATGGCACGTAAAACCGCAGCAGAA	SrpR_Rev_D	ACGAAGACCTACCTTTATTCGAAGGATTTCACCTGTTT
pTet_AK	pTet_For_A	ATGAAGACGTGGAGTCCCTATCAGTGATAGAGATTGAC	pTet_Rev_K	ACGAAGACCTGCATTTCGGTCAGTGCGTCCTGCTGATG
pTet_KB	pTet_For_K	ATGAAGACGTATGCTCCCTATCAGTGATAGAGATTGAC	pTet_Rev_B	ACGAAGACCTAGTATTCGGTCAGTGCGTCCTGCTGATG
pBad_AK	pBad_For_A	ATGAAGACGTGGAGAAGAAACCAATTGTCCATATTGCA	pBad_Rev_K	ACGAAGACCTGCATTATGGAGAAACAGTAGAGAGTTGC
pBad_KB	pBad_For_K	ATGAAGACGTATGCAAGAAACCAATTGTCCATATTGCA	pBad_Rev_B	ACGAAGACCTAGTATATGGAGAAACAGTAGAGAGTTGC
pSrpR_KB	pSrpR_For_K	ATGAAGACGTATGCTTCGTTACCAATTGACAGCTAGCT	pSrpR_Rev_B	ACGAAGACCTAGTAGTTTACAAACAAACAAGCATGTAT
pLmrA_FK	pLmrA_For_F	ATGAAGACGTCGCTTTCGTTACCAATTGACAACTGGTG	pLmrA_Rev_K	ACGAAGACCTGCATAAATATAGTGACTGGTCTATTATC
pBetI_EB	pBet_For_E	ATGAAGACGTGCTTTTCATGGATTCGTTACCAATTGAC	pBetI_Rev_B	ACGAAGACCTAGTAGCTAGCATTATATTGAACGTCCAA
pPhlF_GB	pPhlF_For_G	ATGAAGACGTTGCCTTCGTTACCAATTGACATGATACG	pPhlF_Rev_B	ACGAAGACCTAGTAACCTTAACGATACGGTACGTTTCG
pBM3R1_FB	pBM3R1_For_F	ATGAAGACGTCGCTTTCGTTACCAATTGACGGAATGAA	pBM3R1_Rev_B	ACGAAGACCTAGTAGCTAGCATTATCGGAATGAACGTT

Primer Design for Fusion Proteins

Device Name	Primer	Sequence
C0080_CI	C0080_Rev_I	ACGAAGACCTTAGACAACTTGACGGCTACATCATTCAC
C0040_CI	C0040_Rev_I	ACGAAGACCTTAGACAACTTGACGGCTACATCATTCAC
E0040m_ID	E0040m_For_I	ATGAAGACGTTCTAGAATGCGTAAAGGAGAAGAACTTTTC
E0030_ID	E0030_For_I	ATGAAGACGTTCTAGAATGGTGAGCAAGGGCGAGGAGCTG

Flow Cytometry Data

For the first tandem promoter flow cytometry experiment, we tested pBad-pTet-BCD2-E1010m-B0015 and pTet-pBad-BCD2-E1010m-B0015. We planned on inducing each promoter separately with it's corresponding small molecule (either arabinose or atc) and also planned on inducing both together. We followed the

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