Melbourne/Plan:Gas vesicles

Steps:	
#       Usefull links [[Melbourne/primary Restriction enzymes|(restriction enzymes)]]
#       Sequences:
##      Ncbi genebank AF053765  [[Melbourne/AF053765-pNL26|(pNL26 7371bp)]]  [[Melbourne/AF053765-pNL29|(pNL29 6036bp)]] [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nuccore&id=3089519| (original source)] [[Melbourne/AF053765FASTA| (FASTA seq.)]]
##      pBluescriptIIKS+ [http://www.stratagene.com/products/displayProduct.aspx?pid=267 (stratagene) ][http://www.stratagene.com/vectors/sequences/pbl2ksp_s.txt (sequence) ] [http://www.stratagene.com/vectors/restriction_sites/pbl2ksp_r.txt (restriction map) ][http://www.stratagene.com/vectors/maps/pdf/pBluescript%20II%20KS+_%20webpg.pdf (map) ] [http://www.stratagene.com/manuals/212205.pdf (Manual)] 
##*[[Melbourne/pBluescriptIIKS nospaces|(no spaces sequence 2961bp)]] 
##*HindIII cuts at 719 ,EcoRI cuts at 707 ,PstI cuts at 701  ,Xbal cuts at 677  ,SpeI cuts at 683 
##*[[Melbourne/pBluescriptIIKS PstI HindIII |(no spaces sequence HindIII *AGCTT....CTGCA* PSTI 2947bp)]]
##      pNL26 Plasmid with insert:[[Melbourne/cannon plasmid pNL26 seq| (seq 10308bp)]] [[Melbourne/cannon plasmid pNL26 res map| (res map)]] [[Melbourne/cannon plasmid pNL26 HindIII digest|(digests)]]
###     pNL26 insert PstI--HindIII:[[Melbourne/pNL26 insert|(no spaces sequence 7376bp)]]
##      pNL29 Plasmid with insert:[[Melbourne/cannon plasmid pNL29 seq| (seq 8973bp)]] [[Melbourne/cannon plasmid pNL29 res map| (res map)]] [[Melbourne/cannon plasmid pNL29 HindIII digest| (digests)]]
###     pNL29 insert PstI--HindIII:[[Melbourne/pNL29 insert|(no spaces sequence 6041bp)]]

#	Recovery of genes: (2 days)
##	Recover the plasmid from paper provided into solution.(method)
##	Transform E.Coli strain DH5alpha. Screen with Amp 100ug/ml (as per ref.)(electroporation & heatshock)
##      pick 3 colonies of each
##	Overnight Culture x9(6 above and 3 form agar block provided)
##      Miniprep
##      Produce glycerol stocks
##	Confirm presence in recovered sample using digest.(HindIII)
##      ->Established Supply of Plasmid

#	Induce translation overnight 37deg with IPTG in 100ml plates (method)(2days)
##      Confirm transcription RT-PRC, 
##      Confirm translation (buoyant phenotype method).
##      Confirm translation (Namarski optics (direct interferance contrast microscopy method).

#	Removal of four biobrick like restriction sites all in GvpL.
##	EcoRI [GAATTC] in gvpL (2858)
##	PstI [CTGCAG] in gvpL x 3 (2522,2900,3005)
##	XbaI [TCTAGA] (not present)
##	SpeI [ACTAGT] (not present)

#	Insertion of biobrick required restriction sites by PCR primer modification.(method)
##	Design of primers
##	Primer generation
##	Plasmid extraction from culture
##	PCR 
##	Restriction EcoR1 & Spe1
##	Gel separation
##	Restriction of standard Library death plasmid EcoR1,Spe1.
##	Ligation.
##	Transform host with regulated POPS output
##	Confirm dna, rna , protein (as for A)