US 11,999,959 B2 · Binary vectors and uses of same

Abstract

The invention relates to binary vectors based on compatible and autonomous origins, specifically based on the pBBR1 and RK2 replication origins. These binary vectors are useful for having a wide range of hosts, for their maintenance in Agrobacterium sp. and Escherichia coli, and as a new tool for plant synthetic biology as well as a flexible framework for assembly, transfer and characterization of multiple DNA elements. The binary vectors disclosed are small, preferably less than 3.8 kb in size, stable, include an origin compatible with the most commonly used binary T-DNA vectors, comply with current standards for plant synthetic biology, and allow the administration of multiple T-DNA cassettes by means of the multiplexing of the vectors. The present invention also relates to methods for transferring and expressing nucleic acid sequences using said binary vectors, and to the uses of the same.

Claims20

Dependent claims

A binary vector comprising a nucleic acid sequence comprising in this order:

(a) a T-DNA right border comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:
102 and SEQ ID NO:
115;
and
(b) a sequence portion connecting item (a) with the following item (c);
and
(c) a bacterial terminator;
and
(d) an origin of replication that is functional in at least one bacterial species;
and
(e) a bacterial terminator;
and
(f) a sequence portion connecting item (e) with the following item (g);
and
(g) a T-DNA left border comprising a nucleotide sequence selected from the group consisting of SEQ ID NO:
103, SEQ ID NO:
104 and SEQ ID NO:
116;
and
wherein the bacterial terminator comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:
108, SEQ ID NO:
109, SEQ ID NO:
110 and SEQ ID NO:
111;
and
wherein the sequence of item (b) and item (f) has a size of less than 200 nucleotides.

23 more elements

Dependencies are inferred from the claim text.

Binary vectors and uses of same

Life of this patent

Binary vectors and uses of same

Life of this patent

Abstract

Claims20