Lucigen Corporation

Varigen Biosciences Corporation

Terra Bioworks, Inc.

David Mead

Ronald Godiska

Robert Joseph Stankey

Thomas W Schoenfeld

Nikolai V Ravin

Eric J Steinmetz

Laura P Franz

Vinay K Dhodda

Robert A Difrancesco

Saurabh Sen

Spencer Hermanson

Michele E Auldridge

Disclosed herein are recombinant methods of activating expression of one or more biosynthetic gene clusters comprising more than one gene, the method comprising a recombinant DNA expression vector that possess two opposable inducible promoters that drives expression of a biosynthetic gene cluster exogenously from outside of the cluster to produce polyketides or non-ribosomal peptides in a heterologous host.

Expression vector

Linear vectors derived from bacteriophage ofand host cells suitable for cloning are provided. The linear vectors include a left arm comprising a left telomere and a first selectable marker, a right arm comprising a right telomere and a second selectable marker and a cloning region located between the left arm and the right arm. Optional further components of the vector include transcriptional termination sequences, multiple cloning sites and reporter stuffer regions.

Linear vectors, host cells and cloning methods

Described herein are isolated polynucleotides that encode a fluorescent protein which is at least 80% sequence identical to a protein selected from the group consisting of SEQ. ID. NOS: 2, 4, 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, and 74. Also described are expression constructs containing the polynucleotides, transformed host cells containing the expression constructions, the encoded fluorescent proteins themselves, and methods of using the nucleotides and encoded fluorescent proteins for bioanalytical research.

Lucigen yellow (LucY), a yellow fluorescent protein

A system for ligase-free cloning and/or expressing a target gene is described herein. A preferred version of the invention includes anhost. The host preferably includes a T7 RNA polymerase gene comprising a T7gpl coding sequence, a lacUV5 promoter, and a lac operator. The host preferably further includes a lacI gene comprising a lacI coding sequence with an ATG start codon, a promoter derived from the lacallele, and a translational enhancer derived from a 5' RNA leader sequence of T7 gene 10. The invention further includes a low-copy plasmid vector comprising a T7 promoter a lac operator operationally linked to the T7 promoter. The system is configured to inhibit target gene expression when uninduced and to permit gene expression upon induction by auto-induction.

Host-vector system for cloning and expressing genes

Thermostable viral polymerases exhibiting a combination of activities selected from, proofreading (3'-5&#x2032;) exonuclease activity, nick translating (5&#x2032;-3&#x2032;) nuclease activity, synthetic primer-initiated polymerase activity, nick-initiated polymerase activity, reverse transcriptase activity, strand displacement activity, and/or decreased discrimination against incorporation of nucleotide analogs. Also provided are compositions including the polymerases, polynucleotides encoding the polymerases and methods of using the polymerases.

Thermostable viral polymerases and methods of use

Vector preparations and cloning constructs suitable for use in cloning are provided. Vector preparations are double-stranded DNA molecules having two 3' termini, each terminus having a single base pair overhang that is capable of hybridizing to a single base pair overhang on a double stranded polynucleotide sequence to be cloned. The overhang of the vector preparation is suitably a dCMP and the overhang of the polynucleotide sequence to be cloned is suitably a dGMP. In other embodiments, the overhang of the polynucleotide sequence to be cloned is any ddNTP and the corresponding overhang of the vector preparation is any base that pairs to the ddNTP. The latter embodiment is particularly suited to preparing recombinant molecules having only a single insert. Methods of cloning, methods of preparing libraries of recombinant molecules and kits for carrying out the methods are also provided.

Vectors, kits and methods for cloning DNA

The present invention relates to systems, methods, and compositions for cloning and sequencing insert nucleic acid sequences. In particular, the present invention provides vectors and vector components configured for multiplex cloning, multiplex sequencing, and fixed orientation cloning. The present invention also provides vectors and vector components that allow insert sequences that are deleterious to a host cell to be successfully cloned.

Growing community of inventors

Middleton, WI, United States of America

David Mead