Polymerase enzyme

Polymerase comprising,

• a. an N-terminal 5′-3′nuclease domain,
• i. stemming from Taq polymerase or,
• ii. a polymerase sharing at least 95% amino acid sequence identity with the N-terminal 5′-3′ nuclease domain of Taq polymerase,
• b. an adjacent and linked polymerase domain, stemming from a viral family A polymerase, wherein the polymerase domain stems preferably from,
• 1. JGI20132J14458_100001622 (1607 amino acids;
• SEQ ID NO. 20), or a functional fragment that shares at least 98% amino acid sequence identity thereto, and is altered to comprise the following amino acid changes, Q627N, H751Q, Q752K, and V753K, or
• 2. Ga0186926_122605 (1595 amino acids;
• SEQ ID NO. 21), or a functional fragment that shares at least 98% amino acid sequence identity thereto, and is altered to comprise the following amino acid changes, Q627N, H752Q, Q753K, and V754K, or
• 3. Ga0080008_15802729 (1619 amino acids;
• SEQ ID NO. 22) or a functional fragment that shares at least 98% amino acid sequence identity thereto, and is altered to comprise the following amino acid changes, Q628N, H752Q, Q753K, and L754K, or
• 4. Ga0079997_11796739 (1608 amino acids;
• SEQ ID NO. 23), or a functional fragment that shares at least 98% amino acid sequence identity thereto and is altered to comprise the following amino acid changes, Q627N, H752Q, Q753K, and I754K.

Polymerase comprising,

• a. the amino acid sequence of
• i. SEQ ID NO. 16 (OP-2605) comprising the following additional amino acid changes, Q627N, H752Q, Q753K, and V754K,
• ii. or an amino acid sequence at least 95%, preferably at least 98% identical thereto,
• b. the amino acid sequence of
• i. SEQ ID NO. 15 (OS-1622) comprising the following additional amino acid changes, Q627N, H751Q, Q752K, and V753K,
• ii. or an amino acid sequence at least 90%, preferably at least 95%, more preferably at least 98% identical thereto,
• c. the amino acid sequence of
• i. SEQ ID NO. 17 (CS-2729) comprising the following additional amino acid changes, Q628N, H752Q, Q753K, and L754K, or an amino acid sequence at least 90%, preferably at least 95%, more preferably at least 98% identical thereto, or
• d. the amino acid sequence of
• i. SEQ ID NO. 18 (PS-6739) comprising the following additional amino acid changes, Q627N, H752Q, Q753K, and I754K,
• ii. or an amino acid sequence at least 90%, preferably at least 95%, more preferably at least 98% identical thereto.

Polymerase domain selected from the group of:

• (a) OP-2605 (577 amino acids) according to SEQ ID NO. 25 (derived from Locus tag Ga0186926_122605),
• (b) OS-1622 (576 amino acids) according to SEQ ID NO. 24 (derived from Locus tag JGI20132J14458_100001622),
• (c) CS-2729 (577 amino acids) according to SEQ ID NO. 26 (derived from Locus tag Ga0080008_15802729), or
• (d) PS-6739 (577 amino acids) according to SEQ ID NO. 27 (derived from Locus tag Ga0079997_11796739), or
• (e) polypeptide polymerase domain or functional fragment that shares more than 80%, 85%, 90%, 95% or 99% sequence identity with (a), (b), (c) or (d).