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1. (WO2017095793) TRANSCRIPTION TERMINATOR AND USE THEREOF
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CLAIMS

1. A non-naturally occurring nucleic acid sequence comprising a Y— X— Z stem-loop,

wherein:

Y is a nucleotide sequence of 10 to 30 nucleotides in length;

X is a nucleotide sequence of 3 to 12 nucleotides in length, each nucleotide therein not base pairing with any other nucleotide within X; and

Z is a nucleotide sequence of 10 to 50 nucleotides in length and having at least 70% complementarity to Y.

2. The non-naturally occurring nucleic acid sequence of claim 1, wherein Y has a G/C

content of at most 60%, preferably at most 50%, more preferably at most 40%.

3. The non-naturally occurring nucleic acid sequence of claim 1, wherein Y is engineered to be 5' to X.

4. The non-naturally occurring nucleic acid sequence of claim 1, wherein Y is engineered to be 3' to X.

5. The non-naturally occurring nucleic acid sequence of any one of claims 1-4, wherein Y is

12-18 nucleotides in length, or 14-16 nucleotides in length, or 16-18 nucleotides in length, or 17-19 nucleotides in length, or 15-30 nucleotides in length, or 18-27 nucleotides in length, or 21-24 nucleotides in length, or 24-28 nucleotides in length, or 25-29

nucleotides in length.

6. The non-naturally occurring nucleic acid sequence of any one of claims 1-4, wherein X is 3-8 nucleotides in length, preferably 4-6 nucleotides in length, more preferably 5-6 nucleotides in length.

7. The non-naturally occurring nucleic acid sequence of any one of claims 1-4, wherein Z has the same length as Y and has one or more mismatches with Y.

8. The non-naturally occurring nucleic acid sequence of any one of claims 1-4, wherein Z has a different length than Y and has one or more insertions or deletions compared to Y.

9. The non-naturally occurring nucleic acid sequence of any one of claims 1-4, comprising AAGC.

10. The non-naturally occurring nucleic acid sequence of any one of claims 1-4, comprising CATC.

11. The non-naturally occurring nucleic acid sequence of any one of claims 1-4, having the sequence of SEQ ID NO: 3, 4, or 6.

12. A transcription terminator comprising a first stem-loop and a second stem-loop, wherein the first stem-loop has the non-naturally occurring nucleic acid sequence of any one of claims 1-4, and wherein the first stem-loop is engineered to be 5' to the second stem-loop.

13. The transcription terminator of claim 12, wherein the second stem-loop is a short stem- loop.

14. The transcription terminator of claim 12, wherein the second stem-loop has the non- naturally occurring nucleic acid sequence of any one of claims 1-4.

15. The transcription terminator of claim 12, further comprising a third stem-loop.

16. The transcription terminator of claim 15, wherein the third stem-loop is a short stem-loop.

17. The transcription terminator of claim 15, wherein the third stem-loop has the non- naturally occurring nucleic acid sequence of any one of claims 1-11.

18. The transcription terminator of claim 12, having the sequence of SEQ ID NO: 2 or 5.

19. A vector comprising the transcription terminator of claim 12, wherein the transcription terminator is operably linked to a DNA insert.

20. The vector of claim 19, having the sequence of SEQ ID NO: 1.

21. An engineered cell comprising the vector of claim 19 or 20.

22. A method of engineering a vector, comprising providing the transcription terminator of claim 12 in a vector, wherein the transcription terminator is engineered to operably link to a DNA insert.

23. A method of terminating transcription of a DNA insert, comprising:

a. providing the transcription terminator of claim 12 engineered to operably link to the DNA insert;

b. allow transcription of the DNA insert; and

c. terminate transcription of the DNA insert at the transcription terminator.