IGEM:MIT/2005/Inventory
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File:PlasmidInventory IGEM2005.xls
Strains
- NB115 (pCT-4-4-20) -- 7.02 -80°C
- NB116 (pCT302 S101A) -- 7.02 -80°C
- NB118 (pCT-4M5.3) -- 7.02 -80°C
- MC4100
- Genotype: F- araD139 delta(argF-lac)U169 prsL150 relA1 deoC1 rbsR fthD5301 fruA25 lambda- [s]
Plasmids
[[../Plasmid Info/]]
- pSB1AK3-1 -- as plate, to be made into glycerol stock thursday
- pSB1AC3-1 -- ditto
- pSB1AT3-1 -- ditto
- pSB2K3-1 -- ditto
- File:PCT302.pdf
- [[../PCT302 (aka pCT-4-4-20)/]]-- 7.02 4°C
- [[../PCT302 S101A/]] -- 7.02 4°C
- [[../PCT-4M5.3/]] -- 7.02 4°C
PCR Amplification
- Spin PCR purification kit ()
- Platinum, TAQ polymerase (DNA polymerase for PCR)
- DNTP mix (25mm ea. dntp) (nucleotides for PCR)
Agarose gels
- Agarose 100g (pouring gels)
- 10X TAE buffer (pouring/running gels)
- 1 kb DNA ladder 250µg (molecular weight standard)
- Ethidium Bromide Solution (10mg/mL) (staining DNA)
Cutting/joining DNA fragments
- EcoR1 10,000 u (Restriction Enzyme)
- PstI 10,000 u (Restriction Enzyme)
- SpeI 2500 u (Restriction Enzyme)
- Xba1 3,000 u (Restriction Enzyme)
- T4 DNA Ligase 100,000un (Ligating DNA fragments)
Purifying DNA
- Spin Plasmid Kit, Mini(for minipreps)
- Gel Extraction Kit 50 (purifying from gel slices)
- Spin PCR Purification Kit (purifying PCR products)
Microbiology supplies
- LB amp -- Cold Room
- Amp stock -- -20 Fridge
- VWR Petri Dish (100x15mm, case of 500) (for pouring plates)
- Test Tube, Culture (Polystyrene, 12x75, case of 1000) (special tubes for FACS)
Bio Bricks
pSB1AT3-1
- BBa_J07012 scFv anti-fluorescein [4-4-20]
- BBa_J07013 scFv anti-fluorescein [S101A]
- BBa_J07014 scFv anti-fluorescein [4M5.3]
- BBa_J07017 FecA protein (full length)
pSB1AC3
- BBa_J07021 FecR
- BBa_J07022 FecI
pSB1AK3
- BBa_J07043 J07012.B0015 scFv4-4-20.TT
- BBa_J07044 J07013.B0015 scFvS101A.TT
- BBa_J07045 J07014.B0015 scFv4M5.3.TT
Miniprepped DNA
- R0053 -- Amp -- KBlab -- 4°C
- R0040 -- Amp -- KBlab -- 4°C
- R0011 -- Amp -- KBlab -- 4°C
- [[../Quad part inverters/]]
- E0420 -- Amp -- KBlab -- 4°C
- E0430 -- Amp -- KBlab -- 4°C
- E0840 (rbs:gfp:term)
- B0015 (term)
- R0040 (tetR promoter)
- R0011 (lacI regulated promoter)
- B0030 (Strong RBS)
- J13002 (R0040.B0034)
Streaked out on a plate
- I0500 (pBad/AraC promoter)
Ordered as Stabs
- R0010
- R0051
- J04500
Assemblies Ordered from Registry
BBa_J07036 BBa_J07037 BBa_J07038 BBa_J07039 BBa_J07040 BBa_J07041 BBa_J07042
No Longer Needed:
- I1030
- I1031
- I1032
- I1033
Planning/Building Bio Bricks
- BBa_J07005 [msr] E.coli JM83 alkaline phosphatase gene (PhoA) 1416 bp
- BBa_J07006 [tmp] malE 1191 bp
- BBa_J07007 [reg] ctx promoter 145 bp
- BBa_J07008 [tmp] ToxR w.t. (from Vibrio Cholarae, aa's 1-294) 1160 bp
- BBa_J07009 [reg] ToxR' (aa's 1-210) 630 bp
- BBa_J07010 [reg] ToxR inner (aa's 1-198; cytoplasm + TM) 594 bp
- BBa_J07011 [rpt] ctx promoter :: gfp reporter 871 bp
Primers
Please click link in order to see [[../Sequences of primers specified/]]. They consist of the following:
Ordered/Received:
- Not completely or not received primers are listed in italics
- scFv fwd, rev
- FecA fwd, rev, mid-fwd, mid-rev
- FecA promoter fwd, rev
- MalE fwd, rev
- PhoA fwd, rev, mid1-fwd, mid1-rev, mid2-fwd, mid2-rev
- FecI fwd, rev
- FecR fwd, rev, mid-fwd, mid-rev
- ToxR ATCC fwd, rev
- Linker fwd1
- FecA Promoter
- Fusion
Arrived: 8/4/2005 fwd & rev138 fwd & rev155 fwd & rev176 fwd & rev178 fwd180 fwd & rev527 fwd & rev571 fwd & rev138-141 fwd & rev155-158 fwd & rev176-180 fwd & rev178-182 fwd & rev180-184 fwd & rev521-529 fwd & rev523-531 fwd & rev527-531 fwd & rev567-576 fwd571-574
Specified:
- ToxR (ATCC)
- scFv 3' linker
- scFv fwd flag test (for introducing a start codon)
- FecA knockout with Tetracycline marker
- Fur knockout with Chloramphenicol marker
- FecA'-scFv fusion
- FecA promoter
(n.b: "mid" refers to primers intended to introduce point mutations to take out restriction sites in our parts)
