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Assembly Ordering and Progress
system 1
System 1: glucose-sensing insulin generator (5 task left: 2 PCR and 3 biobrick standard assembly)
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pOmpC |
RBS + TAT
|
INSA |
RBS + TAT
|
INSB |
RBS |
CinR |
RBS |
HSL |
D-term |
duration |
members |
assembly |
status |
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RBS + CinR |
RBS + HSL |
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8/30-8/31 |
翔 |
1st assembly |
9/01, 07
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CinR + HSL (RBS+) |
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9/1-9/4 |
宥,君,威 |
2nd assembly |
9/02, 07
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CinR+HSL(RBS+) + D-term |
9/16-9/27 |
宥,君,翔,pett |
3rd assembly |
9/27 07
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RBS + TATA+ INSA (PCR) |
RBS + TATB+ INSB (PCR) |
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9/28-9/30 |
銘, pett
|
basic part (PCR) |
9/30, 07
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pOmpC + TATA_INSA (failed) |
TATB_INSB + CinR+HSL+D-term (wrong assembly)
|
-10/5 |
pett |
4th assembly |
10/5,7,8 07
|
pOmpC + TATA_INSA |
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-10/11 |
pett |
4th assembly |
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pOmpC + TATA_INS_A + TATB_INS_B
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-10/23 |
|
4th assembly |
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pOmpC + TATA_INS_A + TATB_INS_B + CinR+HSL+D-term |
|
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Final
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system 2
System 2: insulin-sensing insulin destoryer (5 tasks left: 2 PCR and 3 biobrick standard assembly)
|
pCinRHSL |
OmpRBS |
RBS + TATD
|
IDE |
D-term |
duration |
members |
assembly |
status |
|
OmpRBS (PCR) |
RBS + TATD+ IDE (PCR)
|
|
-102 |
銘, 茹, pett
|
basic part (PCR) |
10/2 07
|
pCinRHSL + OmpRBS |
|
|
|
-10/5 |
pett |
1st assembly |
10/5 07
|
pCinRHSL + OmpRBS + TATD_IDE (RBS+) |
|
-10/11 |
pett |
2nd assembly |
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pCinRHSL+OmpRBS+TATD_IDE + D-term |
|
|
Final |
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Promoter assay construct: pOmpC+(RBS+EYFP)
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- our first contruct for promoter activity assay under glucose stimulus
- Insert size: 866bp (XbaI to SpeI)
- Total vector size: 3,019bp
- V-I: 2,153bp
- May be checked with: BtgZI
- 8/27, 2007
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OmpRBS assay construct: pLacIQ + OmpRBS + GFP
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- our first construct for OmpRBS blocking assay under glucose stimulus
- Insert size: 1004bp (XbaI to SpeI)
- Total vector size: 5,425bp
- V-I: 4,421bp
- May be checked with: BtgZI
- 10/24, 2007
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OmpRBS assay construct: pLacIQ + OmpRBS
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- our first construct for OmpRBS blocking assay under glucose stimulus
- Insert size: 120bp (XbaI to SpeI)
- Total vector size: 4,541bp
- V-I: 4,421bp
- May be checked with:
- 12/13, 2007
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System 1: pOmpC + (TATA+INSA) + (TATB+INSB) + (RBS+CinR) + (RBS+HSL) + D-term
Phase 1 of system 1: RBS + CinR
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- Insert size: 790bp
- Total vector size: 2,943bp
- V-I: 2,153bp
- May be checked with: BsaBI
|
Phase 1 of system 1: RBS + HSL
|
- Insert size: 730bp
- Total vector size: 2,883bp
- V-I: 2,153bp
- May be checked with: AgeI
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Phase 1 of system 1: TATA + INSA = TATA_INSA
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- Insert size: 170bp
- PCR product size: 209bp
- May be checked with: RsaI
- There is a extra SpeI site formed due to the addition of one extra stop codon
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Phase 1 of system 1: TATB + INSB = TATB_INSB
|
- Insert size: 220bp
- PCR product size: 233bp
- May be checked with AluI
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Phase 2 of system 1: pOmpC + TATA_INSA
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- Insert size: 292bp
- Total vector size: 2,454bp
- V-I: 2,162bp
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Phase 2 of system 1: (RBS+CinR) + (RBS+HSL) = CinR+HSL
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- Insert size: 1,524bp
- Total vector size: 3,677bp
- V-I: 2153bp
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Phase 3 of system 1: pOmpC + TATA_INSA + TATB_INSB
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- Insert size: 502bp
- Total vector size: 2,655bp
- V-I: 2,153bp
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Phase 3 of system 1: CinR+HSL + D-term = CinR+HSL+D-term
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- Insert size: 1,633bp
- Total vector size: 4,818bp
- V-I: 3,185bp
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Phase 4 of system 1: pOmpC + TATA_INSA + TATB_INSB + CinR+HSL+D-term
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- Insert size: 2,139bp
- Total vector size: 4,292bp
- V-I: 2,153bp
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System 2: pCinRHSL+ OmpRBS + (TATD+IDE) + D-term
Phase 1 of system 2: TATD + IDE = TATD_IDE
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- Insert size: 3,190bp
- PCR product size: 3,203bp
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Phase 1 of system 2: pCinRHSL + OmpRBS
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- Insert size: 308bp
- Total vector size: 2,461bp
- V-I: 2,153bp
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Phase 2 of system 2: pCinRHSL + OmpRBS + TATD_IDE
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- Insert size: 3,488bp
- Total vector size: 5,641bp
- V-I: 2,153bp
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Phase 3 of system 2: pCinRHSL + OmpRBS + TATD_IDE + D-term
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- Insert size: 3,589bp
- Total vector size: 5,742bp
- V-I: 2,153bp
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Basic Constructs
pOmpC in pSB1A2
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- Insert size: 118bp
- Total vector size: 2,271bp
- V-I: 2,153bp
|
pCinRHSL in pSB1A2
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- Insert size: 235bp
- Total vector size: 2,388bp
- V-I: 2,153bp
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RBS in pSB1A2
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- BseR1 can be applied to check the insertion of RBS
- Insert size: 22bp
- Total vector size: 2,175bp
- V-I: 2,153bp
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D-term in pSB1AK3
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- Insert size: 105bp
- Total vector size: 3,290bp
- V-I: 3,185bp
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