Xis from bacteriophage lambda, assembly standard 21
T4 holin gene without a start codon with a stop codon, in BBb format. ---- This part is in BBb Format. It is flanked by BamHI and BglII sites instead of XbaI and SpeI. More information about the BBb Format is available at: [http://openwetware.org/wiki/Template:AndersonLab:BBb_Standard BBb Standard Description Page]
PCR bx3/bx2 on Bjh1301 (685 bp, EcoRI/BamHI/DpnI) Sub into pBca1256 (EcoRI/BamHI) Product is pBca1256-K112001 ---------------------------------------- bx3 Forward biobricking of {<T4 Holin!} ccagtGAATTCatgAGATCTgcagcacctagaatatcattttcgccc bx2 Reverse biobricking of {<T4 Holin!} cgttaGGATCCttatttagcccttcctaatattctggccgcc
{a~xis!} The bacteriophage lambda xis gene ready to have rbs attached and stop codon; assembly stand
This part is in BBb Format. It is flanked by BamHI and BglII sites instead of XbaI and SpeI. More information about the BBb Format is available at: [http://openwetware.org/wiki/Template:AndersonLab:BBb_Standard BBb Standard Description Page]
excisionase from E. coli phage lambda (removes prophage from host genome)
Lambda excisionase coding region with AAV degradation tag and two stop codons
attR2 recombination site-reverse orientation
attR2 recombination site in reverse orientation
This part is in BBb Format. It is flanked by BamHI and BglII sites instead of XbaI and SpeI. More information about the BBb Format is available at: BBb Standard Description Page
{< xis >} The bacteriophage lambda xis gene without start or stop codons; assembly standard 21
{rbs_pelB>} {< xis>} ---- This part is in BBb Format. It is flanked by BamHI and BglII sites instead of XbaI and SpeI. More information about the BBb Format is available at: [http://openwetware.org/wiki/Template:AndersonLab:BBb_Standard BBb Standard Description Page]
Digest (BgIII methylated) K112230 AK, (BamHI methylated) K112203 KC (BamHI/BgIII/XHoI) Product is pBjh1601AC-K112327
{rbs.xis!} The xis gene with native rbs site and stop codon; assembly standard 21
Xis Int temperature sensitive expression cassette
Xis int (with native rbs) under the control of a temperature sensitive promoter. Terminator follows CDS.
{xis>} The lambda xis gene with no stop codon; assembly standard 21
Lambda phage attB site, reverse complement (B'-O-B)
---- This part is in BBb Format. It is flanked by BamHI and BglII sites instead of XbaI and SpeI. More information about the BBb Format is available at: [http://openwetware.org/wiki/Template:AndersonLab:BBb_Standard BBb Standard Description Page]
{rbs.xis.int!} The bacteriophage lambda xis/integrase gene with native rbs and stop codon; assembly
The xis and int genes naturally overlap, so instead of trying to isolate both the xis and int and put them together, I decided to make them one entire basic part.
{xis!} from bacteriophage lambda; assembly standard 21
{pAH57 promoter}{rbs.xis.int!} inserted into K112950 plasmid
This part is inserted just before the attL1 site in this plasmid. This part is in BBb Format. It is flanked by BamHI and BglII sites instead of XbaI and SpeI. More information about the BBb Format is available at: [http://openwetware.org/wiki/Template:AndersonLab:BBb_Standard BBb Standard Description Page]
{pAH57 promoter}{rbs.xis.int!}{rbs.ihfA!}{rbs.ihfB!} inserted into K112995 plasmid
This part is inserted into the K112995 plasmid between ccdB and attR2. This part is in BBb Format. It is flanked by BamHI and BglII sites instead of XbaI and SpeI. More information about the BBb Format is available at: [http://openwetware.org/wiki/Template:AndersonLab:BBb_Standard BBb Standard Description Page]
Biobrick Compatible Lambda Chromosomal Insertion Plasmid
Xis Int & TrfA combined expression cassette
Xis.Int under the control of a temperature sensitive promoter followed by a constitutively expressed TrfA gene.
Xis Int controlled by temperature sensitive
Xis.int (with native rbs) under the comtrol of a temperature sensitive promoter. terminator follows the CDS.
{pAH57 promoter}{rbs.xis.int!}{rbs.ihfA!}{rbs.ihfB!} inserted into K112950 plasmid
This part is inserted into the K112950 plasmid just before attL1. This part is in BBb Format. It is flanked by BamHI and BglII sites instead of XbaI and SpeI. More information about the BBb Format is available at: [http://openwetware.org/wiki/Template:AndersonLab:BBb_Standard BBb Standard Description Page]