Addgene - pKJ1712 Plasmid Data

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Keith Joung Lab Plasmids

This is commonly
requested with

Strain KJ1C

Strain CSH100

pKJ1267 (aka pAC-alphaGal4)

pMLM36 (aka pBR-UV5-GP- FD2)

pAC-Kan-alphaGal4

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Plasmid 21870: pKJ1712

Gene/insert name:		None
Insert size (bp):		Unknown
Vector backbone:		pKJ1712 (Search Vector Database)
Type of vector:		Bacterial expression
Backbone size (bp):		8723
5' Sequencing primer:		lac promoter (List of Sequencing Primers)
3' Sequencing primer:		M13 forward20
Bacteria resistance:		Kanamycin
High or low copy:		Low Copy
Grow in standard E. coli @ 37C:		No
Please specify bacterial strain for growth and growth condition:		Grow in 30ug/mL Kanamycin. This is a low copy number plasmid and therefore we recommend that minipreps be performed from a 10 ml culture.
Sequence:		View sequence
Author's Map:		View map
Plasmid Provided In:		DH5a
Principal Investigator:		Keith Joung
Terms and Licenses:		MTA

Comments: This plasmid is used to construct OPEN B2H selection strains(see Maeder et al., Mol Cel 2008, PMID 18657511). Clone DNA binding sites into this reporter plasmid.

Annotations for pKJ1712:

581-1492: p15A origin of replication / 1664-2415: C-terminal lacI gene fragment (for recombination) / 2862-3673: Kanamycin-resistance gene / 3987-4030: rrnB T1 terminator / 4162-4189: rrnB T2 terminator / 4319-4325: SapI site #1 / 4352-4358: SapI site #2 / 4381-4386: lac promoter -35 box / 4405-4410: lac promoter -10 box / 4417: lac promoter transcription start point / 4452-4454: HIS3 gene start codon / 5112-5114: HIS3 gene stop codon / 5162-5164: aadA gene start codon / 5951-5953: aadA gene stop codon / 5960-6576: lacZ gene fragment (for recombination) / 6833-6835: sacB stop codon (rev comp strand) / 6833-8254: sacB coding sequence (rev comp strand) / 8252-8254: sacB start codon (rev comp strand)

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Click on map to enlarge

Selected features

CAT/CamR(660-456)

219 - 16

p15A_origin

581 - 1492

ORF frame 2

3673 - 2858

KanR2

3673 - 2858

rrnB_terminator

3888 - 4021

rrnB_T1_terminator

3987 - 4030

rrnB_T2_terminator

4162 - 4189

lac_promoter

4381 - 4410

M13_pUC_rev_primer

4421 - 4443

ORF frame 3

4335 - 5114

HIS3

4452 - 5111

ORF frame 2

5162 - 5953

spect(variant)

5171 - 5953

M13_forward20_primer

5977 - 5961

M13_pUC_fwd_primer

5992 - 5970

lacZ_a

5970 - 6113

CAT/CamR(144-0)

6804 - 6661

ORF frame 2

8254 - 6833

sacB_neg

8691 - 6820

Unique restriction sites

XbaI		1424
ApaI		1894
EcoRV		2133
NarI		2323
NruI		3584
XhoI		3639
EcoRI		4300
NotI		4327
EagI		4327
SalI		4413
NdeI		4642
PstI		5138
BamHI		5140
ApaLI		5435
NcoI		6801
StuI		8012

Please acknowledge the principal investigator if you use this plasmid in a publication.

Also, please include the text "Addgene plasmid 21870" in your Materials and Methods section. This information allows Addgene to create a link from the plasmid page to your publication.