METHODS, SYSTEMS AND COMPOSITIONS REGARDING MULTIPLEX CONSTRUCTION PROTEIN AMINO-ACID SUBSTITUTIONS AND IDENTIFICATION OF SEQUENCE-ACTIVITY RELATIONSHIPS, TO PROVIDE GENE REPLACEMENT SUCH AS WITH TAGGED MUTANT GENES, SUCH AS VIA EFFICIENT HOMOLOGOUS RECOMBINATION
A method is provided to construct libraries of nucleic acids that comprise a plurality of mutations, each of which may be identified with particularity, such as following a selection or screening.
1 . A method to evaluate and/or identify biological nucleic acid and polypeptide constructs comprising:
(a) obtaining or preparing a first library of nucleic acid sequence variants;
(b) obtaining or preparing a second library for which additional components are attached to members of the first library;
(c) subjecting a second library to a desired protocol to enrich a subset thereof based on one or more selections or screens; and
(d) identifying, sequencing, and/or quantifying one or more of the subset for its nucleic acid sequence variants.
2 . The method of claim 1 , additionally comprising attaching barcode sequences as a further additional component, thereby providing for identification of specific variants.
3 . The method of claim 1 , wherein the template priming sequences function also as the barcode sequences.
4 . The method of claim 2 , wherein the template priming sequences function also as the barcode sequences.
5 . A polypeptide sequence or a nucleic acid sequence identified based on the method of claim 1 .
6 . A polypeptide sequence or a nucleic acid sequence identified based on the method of claim 2 .
7 . A polypeptide sequence or a nucleic acid sequence identified based on the method of claim 3 .
8 . A polypeptide sequence or a nucleic acid sequence identified based on the method of claim 4 .
9 . A system for identifying a selected for or screened for sequence comprising an apparatus practicing the method of claim 1 .
10 . A system for identifying a selected for or screened for sequence comprising an apparatus practicing the method of claim 2 .
11 . A system for identifying a selected for or screened for sequence comprising an apparatus practicing the method of claim 3 .
12 . A system for identifying a selected for or screened for sequence comprising an apparatus practicing the method of claim 4 .