Nucleoside triphosphate transporter and uses thereof
Disclosed herein are proteins, methods, cells, engineered microorganisms, and kits for generating a modified nucleoside triphosphate transporter from Phaeodactylum tricornutum . Also disclosed herein proteins, methods, cells, engineered microorganisms, and kits for production of a nucleic acid molecule that comprises an unnatural nucleotide utilizing a modified nucleoside triphosphate transporter from Phaeodactylum tricornutum.
1. A recombinant nucleoside triphosphate transporter having an amino acid sequence selected from SEQ ID NO: 4, SEQ ID NO: 6, and SEQ ID NO: 8.
2. The recombinant nucleoside triphosphate transporter of claim 1 , wherein the recombinant nucleoside triphosphate transporter is encoded by an extrachromosomal nucleic acid molecule.
3. The recombinant nucleoside triphosphate transporter of claim 2 , wherein the extrachromosomal nucleic acid molecule comprises a nucleic acid sequence provided in SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 5, or SEQ ID NO: 7.
4. A prokaryotic cell comprising the recombinant nucleoside triphosphate transporter of claim 1 .
5. The prokaryotic cell of claim 4 , wherein the prokaryotic cell is an engineered cell.
6. The prokaryotic cell of claim 4 , wherein the prokaryotic cell is E. coli.
7. The prokaryotic cell of claim 4 , further comprising an unnatural nucleoside triphosphate transported into the prokaryotic cell using the recombinant nucleoside triphosphate transporter, wherein the unnatural nucleoside triphosphate comprises a base selected from the group consisting of:
8. The prokaryotic cell of claim 7 , wherein the unnatural nucleoside comprises a deoxyribose moiety.
9. The prokaryotic cell of claim 7 , wherein the unnatural nucleoside comprises a ribose moiety.
10. The prokaryotic cell of claim 4 , further comprising an unnatural base pair (UBP) comprising a first unnatural nucleoside and a second unnatural nucleoside, wherein the first unnatural nucleoside comprises a base that is
and the second unnatural nucleoside comprises a base selected from the group consisting of
11. The prokaryotic cell of claim 10 , wherein the second unnatural nucleoside comprises a base that is
12. The prokaryotic cell of claim 4 , wherein the prokaryotic cell is isolated and purified.
13. A nucleic acid molecule encoding the recombinant nucleoside triphosphate transporter of claim 1 .
14. The recombinant nucleoside triphosphate transporter of claim 1 , wherein the recombinant nucleoside triphosphate transporter has amino acid sequence SEQ ID NO: 4.
15. The recombinant nucleoside triphosphate transporter of claim 1 , wherein the recombinant nucleoside triphosphate transporter has amino acid sequence SEQ ID NO: 6.
16. The recombinant nucleoside triphosphate transporter of claim 1 , wherein the recombinant nucleoside triphosphate transporter has amino acid sequence SEQ ID NO: 8.
17. The prokaryotic cell of claim 4 , wherein the recombinant nucleoside triphosphate transporter has amino acid sequence SEQ ID NO: 4.
18. The prokaryotic cell of claim 4 , wherein the recombinant nucleoside triphosphate transporter has amino acid sequence SEQ ID NO: 6.
19. The prokaryotic cell of claim 4 , wherein the recombinant nucleoside triphosphate transporter has amino acid sequence SEQ ID NO: 8.
20. The prokaryotic cell of claim 6 , wherein the recombinant nucleoside triphosphate transporter has amino acid sequence SEQ ID NO: 4.
21. The prokaryotic cell of claim 6 , wherein the recombinant nucleoside triphosphate transporter has amino acid sequence SEQ ID NO: 6.
22. The prokaryotic cell of claim 6 , wherein the recombinant nucleoside triphosphate transporter has amino acid sequence SEQ ID NO: 8.
23. The prokaryotic cell of claim 10 , wherein the cell is E. coli.
24. The prokaryotic cell of claim 23 , wherein the recombinant nucleoside triphosphate transporter has amino acid sequence SEQ ID NO: 4.
25. The prokaryotic cell of claim 23 , wherein the recombinant nucleoside triphosphate transporter has amino acid sequence SEQ ID NO: 6.
26. The prokaryotic cell of claim 23 , wherein the recombinant nucleoside triphosphate transporter has amino acid sequence SEQ ID NO: 8.
27. The prokaryotic cell of claim 11 , wherein the cell is E. coli.
28. The prokaryotic cell of claim 27 , wherein the recombinant nucleoside triphosphate transporter has amino acid sequence SEQ ID NO: 4.
29. The prokaryotic cell of claim 27 , wherein the recombinant nucleoside triphosphate transporter has amino acid sequence SEQ ID NO: 6.
30. The prokaryotic cell of claim 27 , wherein the recombinant nucleoside triphosphate transporter has amino acid sequence SEQ ID NO: 8.