Team:KU Seoul/Details

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The Experiments

Backbone Plasmids
- Plasmid pSB3C5 with BBa_J04450 : 2009 Kit Plate 1 [5C]
- Plasmid pSB3T5 with BBa_J04450 : 2009 Kit Plate 1 [9C]
Promoters
- Promoter arsR [ParsR], zntA [Pznt] and yodA [PyodA](ref) originated from genomic DNA of Escherichia coli XL-1 Blue
Protein Coding Sequences
- Green fluorescent protein(BBa_E0044) : 2009 Kit Plate 1 [14G]
- Red fluorescent protein(BBa_E1010) : 2009 Kit Plate 1 [18F]
- Aryl acylamidase protein(ref) : new biological part
Primers

Template	Primer	Sequence	Length(bp)
Plasmid pSB3C5	pSB3C5_F	gctgctgttTAATAAtactagtagcggccgctgc	34
Plasmid pSB3C5	pSB3C5(+Pars)_R	taataggtgtgaattttgagttggCtctagaagcggccgcga	42
Plasmid pSB3T5	pSB3T5_F	gacgaaaactacgctgctgctgttTAATAAtactagtagcggccgctgc	49
Plasmid pSB3T5	pSB3T5_R	GTCGGCAATATGAAGctctagaagcggccgcga	33
Promoter yodA	PyodA_F	cggccgcttctagagCTTCATATTGCCGACAAAGTACG	38
Promoter yodA	PyodA_R	ATGTGACTTACCCATaacAGTTTCCTCCAGAGTCATTG	38
Green fluorescent protein	GR(+Pars)_F	aattcacacctattaccttcctctgcacttacacattcgttaagtcatatATGc	78
	GR(+Pars)_F	gtaaaggagaagaacttttcactg	78
	GR(+Pznt)_R	ctggagtcgactccagagtcaagttttatcagagatacagcgagcggacg	84
	GR(+Pznt)_R	TCTAGTttattaaacagcagcagcgtagttttcg	84
Red fluorescent protein	RF(+Pznt)_F	tgataaaacttgactctggagtcgactccagagtgtatccttcggttaatATG	75
	RF(+Pznt)_F	gcttcctccgaagacgttatca	75
	RF(+AAV tag)_R	TTATTAaacagcagcagcgtagttttcgtcgtttgctgcAgcaccggtgg	59
	RF(+AAV tag)_R	agtgacgac	59
Aryl acylamidase	AMD_F	CTGGAGGAAACTGTTatgGGTAAGTCACATTCGCCAG	37
	AMD(+AAV tag)_R	TTATTAaacagcagcagcgtagttttcgtcgtttgctgcCAGGGGC	55
	AMD(+AAV tag)_R	CGTCCGGCG	55

[090914]

Streaking of E. coli XL-1 Blue
Preparation of LB plate (containing 100μg/ml ampicillin, 12.5μg/ml tetracycline, 50μg/ml kanamycin and 25μg/ml chloramphenicol of final concentration, respectively) & dry in 37℃ incubator for 12 hours
Inoculation of E. coli DH5a to 5ml LB broth for preparation of competent cell
Design of cloning primers & ordering the primers