DETAILED CORRESPONDENCE
Application Status
1. The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
2. Applicants’ amendment to the claims filed on 12/29/2025 in response to the restriction requirement mailed on 11/03/2025 is acknowledged. This listing of claims replaces all prior listings of claims in the application.
3. Claims 1-8 and 10-21 are pending.
Election/Restrictions
4. Applicant’s election without traverse of Group I, claims 1-8 and 12-20 in the reply filed on 12/29/2025 is acknowledged.
5. Claims 10-11 and 21 stand withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 12/29/2025.
Claims 1-8 and 12-20 are pending and examined on the merits.
Priority
6. Acknowledgement is made of applicant’s claim for foreign priority under 35 U.S.C. 119(a)-(d) to China Patent Application No. CN202011085981.6, filing date 10/12/2020. The certified copy has been filed in the present application, filed on 04/12/2023.
Information Disclosure Statement
7. The IDS filed on 05/05/2023 has been considered by the examiner and a copy of the Form PTO/SB/08 is attached to the office action.
Drawings
8. The Drawings filed on 04/12/2023 are acknowledged and accepted by the examiner.
Claim Rejections - 35 USC § 112(b)
9. The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
10. Claims 7 and 8 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Regarding claims 7 and 8, there is insufficient antecedent basis for the limitation “the sequence between the transposon 3’ terminal repeat sequence…” and furthermore, it is unclear what sequence between the transposon 3’ terminal repeat sequence the limitation is referring to. It is suggested that applicants clarify the meaning of the claims.
Claim Rejections - 35 USC § 103
11. The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
12. Claim(s) 1-8 and 12-20 is/are rejected under 35 U.S.C. 103 as being unpatentable over Qian et al. (US Patent Application Publication 2018/0265890 A1; cited on PTO-892 mailed on 11/03/2025) in view of Mossine et al. (PLoS One, 2013; examiner cited) and Gasanov et al. (Acta Natural, 2015; examiner cited).
13. Claims 1-6 and 12-17 are drawn to a nucleic acid construct, comprising or consisting of the following elements: a transposon 3’ terminal repeat sequence, a first polyA sequence, an insulator sequence with transcription termination function, a transposon 5’ terminal repeat sequence.
Claims 7-8 and 18-20 are drawn to a host cell comprising: (1) the nucleic acid construct according to claim 1, and/or (2) the sequence between the transposon 3’ terminal repeat sequence and the transposon 5’ terminal repeat sequence of the nucleic acid construct according to claim 1.
14. With respect to claim 1, Qian et al. teach a nucleic acid construct comprising a transposon 3’ terminal repeat sequence, a first polyA signal sequence, and a transposon 5’ terminal repeat sequence [see Abstract; paragraphs 0011-0029; 0069].
With respect to claim 2, Qian et al. teach the nucleic acid construct comprising a transposon 3’ terminal repeat sequence, a first polyA signal sequence, a transposon 5’ terminal repeat sequence, a transposase encoding sequence and a promoter controlling the expression of the transposase [see Abstract; paragraphs 0011-0029; 0069].
With respect to claim 3, Qian et al. teach the nucleic acid construct comprising a transposon 3’ terminal repeat sequence, a multiple cloning insertion site, a first polyA signal sequence, a transposon 5’ terminal repeat sequence, a transposase encoding sequence and a promoter controlling the expression of the transposase [see Abstract; paragraphs 0011-0029; 0069].
With respect to claim 4, Qian et al. teach the nucleic acid construct wherein the orientation of the expression cassette of the transposase is opposite to the orientation of the expression cassette of the exogenous gene [see Abstract], wherein the transposon 3’ terminal repeat and 5’ terminal repeat is a PiggyBac transposon terminal repeat [see paragraph 0035], the transposase is a PiggyBac transposase [see paragraph 0035], the promoter is a CMV promoter, SV40 promoter, EF1a promoter, HSP70 promoter, PGK-1 promoter, b-actin promoter and GRP78 promoter [see paragraph 0046], and the transposase coding sequence contains or is operably linked to a single copy or multiple copies of a nuclear localization signal coding sequence [see paragraph 0041].
With respect to claim 5, Qian et al. teach the construct wherein the transposon 3’ terminal repeat sequence is 100% identical to SEQ ID NO: 1 [see alignment attached as APPENDIX A], the transposon 5’ terminal repeat sequence is 100% identical to SEQ ID NO: 6 [see alignment attached as APPENDIX A], the PiggyBac transposase is as shown in SEQ ID NO: 36 [see alignment attached as APPENDIX A], and the nuclear localization signal is a c-myc nuclear localization signal [see paragraph 0041]. The recitation of “as shown in” in reference to a SEQ ID NO is sufficiently broad to encompass fragments of said sequence such that the interpretation of said sequence need only share at least two contiguous nucleotides with the claimed sequence.
With respect to claim 6, Qian et al. teach the nucleic acid construct wherein the nucleic acid construct comprises a sequence as shown in SEQ ID NO: 10 or 14 [see alignment attached as APPENDIX B]. The recitation of “a sequence as shown in” in reference to a SEQ ID NO is sufficiently broad to encompass fragments of said sequence such that the interpretation of said sequence need only share at least two contiguous nucleotides with the claimed sequence.
With respect to claim 7, Qian et al. teach a host cell comprising the nucleic acid construct [see paragraph 0055].
With respect to claim 8, Qian et al. teach a pharmaceutical composition (medicament) comprising the nucleic acid construct and a pharmaceutically acceptable excipient [see paragraphs 0058-0060].
With respect to claim 12, Qian et al. teach the nucleic acid construct comprising a transposon 3’ terminal repeat sequence, a multiple cloning insertion site, a first polyA signal sequence, a transposon 5’ terminal repeat sequence, a transposase encoding sequence, a promoter controlling the expression of the transposase, and exogenous gene of interest [see Abstract; paragraphs 0011-0029; 0062; 0069].
With respect to claim 13, Qian et al. teach the construct wherein the transposase coding sequence and promoter controlling the expression of the transposase are outside the region between the 3’ and 5’ terminal repeat sequence [see paragraph 0017].
With respect to claim 14, Qian et al. teach the construct wherein the nucleic acid construct further comprises a multiple cloning insertion site and an exogenous gene of interest [see Abstract; paragraphs 0011-0029; 0062; 0069].
With respect to claim 15, Qian et al. teach the nucleic acid construct wherein the coding sequence of the PiggyBac transposase is as shown in SEQ ID NO: 7 and the nuclear localization signal has the sequence that is 100% identical to SEQ ID NO: 35 [see alignment attached as APPENDIX C]. The recitation of “as shown in” in reference to a SEQ ID NO is sufficiently broad to encompass fragments of said sequence such that the interpretation of said sequence need only share at least two contiguous nucleotides with the claimed sequence.
With respect to claim 16, Qian et al. teach the nucleic acid construct wherein the construct is a recombinant vector [see paragraph 0050].
With respect to claim 17, Qian et al. teach the nucleic acid construct wherein the nucleic acid construct is a recombinant cloning vector or a recombinant expression vector [see paragraphs 0050-0051].
With respect to claim 18, Qian et al. teach the host cell wherein the host cell is a mammalian cell [see paragraph 0055].
With respect to claim 19, Qian et al. teach the host cell wherein the host cells are immune cells, Jurkat cell, K562 cell, embryonic stem cell, tumor cell, HEK293 cells, and CHO cell [see paragraph 0055].
With respect to claim 20, Qian et al. teach the host cell wherein the immune cell is a T cell [see paragraph 0055].
However, Qian et al. does not teach the nucleic acid construct comprising an insulator sequence with transcription termination function and the sequential order of the construct as set forth in claim 3.
Mossine et al. teach a piggyBac transposon based reporter expression system wherein the most efficient and stable reporter activity was observed for combinations of transposon inverted terminal repeats and one or two cHS4 core insulators flanking the reporter construct with no detectable silencing over 10 months of continuous cell culture [see Abstract; p. 6, column 2 to p. 8, column 1], wherein the construct sequentially comprised a 3’ transposon terminal repeat, an insulator sequence and 5’- terminal repeat [see Figure 1].
Gasanov et al. teach that mammalian cell lines are widely used to produce recombinant proteins [see Abstract]. Gasanov et al. teach the use of transcription terminators from the SV40 virus, as well as human b- and g- globin genes, to prevent transcription through the transgene. The transcription terminators were shown to increase and stabilize the expression of the EGFP reporter gene in CHO cells and the combination of insulator and terminators provided an additive effect, which enhances and stabilizes transgene expression [see Abstract; p. 77-78, p. 80, column 2; Figure 2].
Before the effective filing date of the claimed invention, it would have been obvious for one of ordinary skill in the art to combine the teachings of Qian et al., Mossine et al. and Gasanov et al. to include insulator sequence with transcription termination function in the constructs of Qian et al. because Qian et al. teach nucleic acid construct comprising a transposon 3’ terminal repeat sequence, a first polyA signal sequence, and a transposon 5’ terminal repeat sequence for stable transfection and expression of exogenous genes in mammalian cells. Mossine et al. teach combinations of transposon inverted terminal repeats and one or two cHS4 core insulators flanking the reporter construct result in stable transformations and Gasanov et al. teach the combination of insulator and terminators provided an additive effect, which enhances and stabilizes transgene expression. One of ordinary skill in the art would have had a reasonable expectation of success, a reasonable level of predictability and would have been motivated to combine the teachings of Qian et al., Mossine et al. and Gasanov et al. because Mossine et al. acknowledges that combinations of transposon inverted terminal repeats and one or two cHS4 core insulators flanking the reporter construct result in stable transformations and Gasanov et al. acknowledges the combination of insulator and terminators provided an additive effect, which enhances and stabilizes transgene expression. Furthermore, it has been held that rearranging parts of an invention involves only routine skill in the art. In re Japikse, 86 USPQ 70. Therefore, the above invention would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the claimed invention.
Conclusion
15. Status of the claims:
Claims 1-8 and 10-21 are pending.
Claims 10-11 and 21 stand withdrawn pursuant to 37 CFR 1.142(b).
Claims 1-8 and 12-20 are rejected.
No claims are in condition for an allowance.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to PAUL J HOLLAND whose telephone number is (571)270-3537. The examiner can normally be reached Monday to Friday from 8AM to 5PM.
Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Manjunath Rao can be reached at 571-272-0939. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
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/PAUL J HOLLAND/Primary Examiner, Art Unit 1656
APPENDIX A
Qian et al. with SEQ ID NO: 1
ALIGNMENT:
Query Match 100.0%; Score 67; Length 67;
Best Local Similarity 100.0%;
Matches 67; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 TTAACCCTAGAAAGATAATCATATTGTGACGTACGTTAAAGATAATCATGCGTAAAATTG 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 TTAACCCTAGAAAGATAATCATATTGTGACGTACGTTAAAGATAATCATGCGTAAAATTG 60
Qy 61 ACGCATG 67
|||||||
Db 61 ACGCATG 67
Qian et al. with SEQ ID NO: 6
ALIGNMENT:
Query Match 100.0%; Score 40; Length 40;
Best Local Similarity 100.0%;
Matches 40; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 GCATGCGTCAATTTTACGCAGACTATCTTTCTAGGGTTAA 40
||||||||||||||||||||||||||||||||||||||||
Db 1 GCATGCGTCAATTTTACGCAGACTATCTTTCTAGGGTTAA 40
Qian et al. with SEQ ID NO: 36
Query Match 97.2%; Score 3137; DB 1; Length 604;
Best Local Similarity 98.8%;
Matches 597; Conservative 0; Mismatches 7; Indels 0; Gaps 0;
Qy 1 MGKPKKIKTEDGSSLDDEHILSALLQSDDELVGEDSDSEVSDHVSEDDVQSDTEEAFIDE 60
|| | ||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MGPAAKRVKLDGSSLDDEHILSALLQSDDELVGEDSDSEVSDHVSEDDVQSDTEEAFIDE 60
Qy 61 VHEVQPTSSGSEILDEQNVIEQPGSSLASNRILTLPQRTIRGKNKHCWSTSKPTRRSRVS 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 VHEVQPTSSGSEILDEQNVIEQPGSSLASNRILTLPQRTIRGKNKHCWSTSKPTRRSRVS 120
Qy 121 ALNIVRSQRGPTRMCRNIYDPLLCFKLFFTDEIISEIVKWTNAEISLKRRESMTSATFRD 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 ALNIVRSQRGPTRMCRNIYDPLLCFKLFFTDEIISEIVKWTNAEISLKRRESMTSATFRD 180
Qy 181 TNEDEIYAFFGILVMTAVRKDNHMSTDDLFDRSLSMVYVSVMSRDRFDFLIRCLRMDDKS 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 TNEDEIYAFFGILVMTAVRKDNHMSTDDLFDRSLSMVYVSVMSRDRFDFLIRCLRMDDKS 240
Qy 241 IRPTLRENDVFTPVRKIWDLFIHQCIQNYTPGAHLTIDEQLLGFRGRCPFRVYIPNKPSK 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 IRPTLRENDVFTPVRKIWDLFIHQCIQNYTPGAHLTIDEQLLGFRGRCPFRVYIPNKPSK 300
Qy 301 YGIKILMMCDSGTKYMINGMPYLGRGTQTNGVPLGEYYVKELSKPVHGSCRNITCDNWFT 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 YGIKILMMCDSGTKYMINGMPYLGRGTQTNGVPLGEYYVKELSKPVHGSCRNITCDNWFT 360
Qy 361 SIPLAKNLLQEPYKLTIVGTVRSNKREIPEVLKNSRSRPVGTSMFCFDGPLTLVSYKPKP 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 SIPLAKNLLQEPYKLTIVGTVRSNKREIPEVLKNSRSRPVGTSMFCFDGPLTLVSYKPKP 420
Qy 421 AKMVYLLSSCDEDASINESTGKPQMVMYYNQTKGGVDTLDQMCSVMTCSRKTNRWPMALL 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 AKMVYLLSSCDEDASINESTGKPQMVMYYNQTKGGVDTLDQMCSVMTCSRKTNRWPMALL 480
Qy 481 YGMINIACINSFIIYSHNVSSKGEKVQSRKKFMRNLYMGLTSSFMRKRLEAPTLKRYLRD 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 YGMINIACINSFIIYSHNVSSKGEKVQSRKKFMRNLYMGLTSSFMRKRLEAPTLKRYLRD 540
Qy 541 NISNILPKEVPGTSDDSTEEPVMKKRTYCTYCPSKIRRKASASCKKCKKVICREHNIDMC 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 NISNILPKEVPGTSDDSTEEPVMKKRTYCTYCPSKIRRKASASCKKCKKVICREHNIDMC 600
Qy 601 QSCF 604
||||
Db 601 QSCF 604
APPENDIX B
Qian et al. with SEQ ID NO: 10
Query Match 68.6%; Score 1804.6; Length 2760;
Best Local Similarity 96.8%;
Matches 1862; Conservative 0; Mismatches 34; Indels 27; Gaps 1;
Qy 637 AAAATAAAGATCTTTTATTGCATGCGTCAATTTTACGCAGACTATCTTTCTAGGGTTAAA 696
|| | | | | | | |||||||||||||||||||||||||||||||||||||||||
Db 330 AACCTCTACAAATGTGGTAGCATGCGTCAATTTTACGCAGACTATCTTTCTAGGGTTAAA 389
Qy 697 TCGATTTAGTCCAACTTGACCCTCTTGGCAGCAGGGAAGCAGCTCTGGCACATGTCGATG 756
|||||||| |||||||||||||||||||||||||
Db 390 TCGATTTA---------------------------GAAGCAGCTCTGGCACATGTCGATG 422
Qy 757 TTGTGCTCGCGGCAGATCACCTTCTTGCACTTCTTGCAGCTGGCGCTGGCCTTGCGGCGG 816
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 423 TTGTGCTCGCGGCAGATCACCTTCTTGCACTTCTTGCAGCTGGCGCTGGCCTTGCGGCGG 482
Qy 817 ATCTTGCTGGGGCAGTAGGTGCAGTAGGTGCGCTTCTTCATCACGGGCTCCTCGGTGCTG 876
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 483 ATCTTGCTGGGGCAGTAGGTGCAGTAGGTGCGCTTCTTCATCACGGGCTCCTCGGTGCTG 542
Qy 877 TCGTCGCTGGTGCCGGGCACCTCCTTGGGCAGGATGTTGCTGATGTTGTCGCGCAGGTAG 936
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 543 TCGTCGCTGGTGCCGGGCACCTCCTTGGGCAGGATGTTGCTGATGTTGTCGCGCAGGTAG 602
Qy 937 CGCTTCAGGGTGGGGGCCTCCAGGCGCTTGCGCATGAAGCTGCTGGTCAGGCCCATGTAC 996
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 603 CGCTTCAGGGTGGGGGCCTCCAGGCGCTTGCGCATGAAGCTGCTGGTCAGGCCCATGTAC 662
Qy 997 AGGTTGCGCATGAACTTCTTGCGGCTCTGCACCTTCTCGCCCTTGCTGCTCACGTTGTGG 1056
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 663 AGGTTGCGCATGAACTTCTTGCGGCTCTGCACCTTCTCGCCCTTGCTGCTCACGTTGTGG 722
Qy 1057 CTGTAGATGATGAAGCTGTTGATGCAGGCGATGTTGATCATGCCGTACAGCAGGGCCATG 1116
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 723 CTGTAGATGATGAAGCTGTTGATGCAGGCGATGTTGATCATGCCGTACAGCAGGGCCATG 782
Qy 1117 GGCCAGCGGTTGGTCTTGCGGCTGCAGGTCATCACGCTGCACATCTGGTCCAGGGTGTCC 1176
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 783 GGCCAGCGGTTGGTCTTGCGGCTGCAGGTCATCACGCTGCACATCTGGTCCAGGGTGTCC 842
Qy 1177 ACGCCGCCCTTGGTCTGGTTGTAGTACATCACCATCTGGGGCTTGCCGGTGCTCTCGTTG 1236
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 843 ACGCCGCCCTTGGTCTGGTTGTAGTACATCACCATCTGGGGCTTGCCGGTGCTCTCGTTG 902
Qy 1237 ATGCTGGCGTCCTCGTCGCAGCTGCTCAGCAGGTACACCATCTTGGCGGGCTTGGGCTTG 1296
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 903 ATGCTGGCGTCCTCGTCGCAGCTGCTCAGCAGGTACACCATCTTGGCGGGCTTGGGCTTG 962
Qy 1297 TAGCTCACCAGGGTCAGGGGGCCGTCGAAGCAGAACATGCTGGTGCCCACGGGGCGGCTG 1356
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 963 TAGCTCACCAGGGTCAGGGGGCCGTCGAAGCAGAACATGCTGGTGCCCACGGGGCGGCTG 1022
Qy 1357 CGGCTGTTCTTCAGCACCTCGGGGATCTCGCGCTTGTTGCTGCGCACGGTGCCCACGATG 1416
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1023 CGGCTGTTCTTCAGCACCTCGGGGATCTCGCGCTTGTTGCTGCGCACGGTGCCCACGATG 1082
Qy 1417 GTCAGCTTGTAGGGCTCCTGCAGCAGGTTCTTGGCCAGGGGGATGCTGGTGAACCAGTTG 1476
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1083 GTCAGCTTGTAGGGCTCCTGCAGCAGGTTCTTGGCCAGGGGGATGCTGGTGAACCAGTTG 1142
Qy 1477 TCGCAGGTGATGTTGCGGCAGCTGCCGTGCACGGGCTTGCTCAGCTCCTTCACGTAGTAC 1536
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1143 TCGCAGGTGATGTTGCGGCAGCTGCCGTGCACGGGCTTGCTCAGCTCCTTCACGTAGTAC 1202
Qy 1537 TCGCCCAGGGGCACGCCGTTGGTCTGGGTGCCGCGGCCCAGGTAGGGCATGCCGTTGATC 1596
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1203 TCGCCCAGGGGCACGCCGTTGGTCTGGGTGCCGCGGCCCAGGTAGGGCATGCCGTTGATC 1262
Qy 1597 ATGTACTTGGTGCCGCTGTCGCACATCATCAGGATCTTGATGCCGTACTTGCTGGGCTTG 1656
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1263 ATGTACTTGGTGCCGCTGTCGCACATCATCAGGATCTTGATGCCGTACTTGCTGGGCTTG 1322
Qy 1657 TTGGGGATGTACACGCGGAAGGGGCAGCGGCCGCGGAAGCCCAGCAGCTGCTCGTCGATG 1716
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1323 TTGGGGATGTACACGCGGAAGGGGCAGCGGCCGCGGAAGCCCAGCAGCTGCTCGTCGATG 1382
Qy 1717 GTCAGGTGGGCGCCGGGGGTGTAGTTCTGGATGCACTGGTGGATGAACAGGTCCCAGATC 1776
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1383 GTCAGGTGGGCGCCGGGGGTGTAGTTCTGGATGCACTGGTGGATGAACAGGTCCCAGATC 1442
Qy 1777 TTGCGCACGGGGGTGAACACGTCGTTCTCGCGCAGGGTGGGGCGGATGCTCTTGTCGTCC 1836
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1443 TTGCGCACGGGGGTGAACACGTCGTTCTCGCGCAGGGTGGGGCGGATGCTCTTGTCGTCC 1502
Qy 1837 ATGCGCAGGCAGCGGATCAGGAAGTCGAAGCGGTCGCGGCTCATCACGCTCACGTACACC 1896
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1503 ATGCGCAGGCAGCGGATCAGGAAGTCGAAGCGGTCGCGGCTCATCACGCTCACGTACACC 1562
Qy 1897 ATGCTCAGGCTGCGGTCGAACAGGTCGTCGGTGCTCATGTGGTTGTCCTTGCGCACGGCG 1956
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1563 ATGCTCAGGCTGCGGTCGAACAGGTCGTCGGTGCTCATGTGGTTGTCCTTGCGCACGGCG 1622
Qy 1957 GTCATCACCAGGATGCCGAAGAAGGCGTAGATCTCGTCCTCGTTGGTGTCGCGGAAGGTG 2016
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1623 GTCATCACCAGGATGCCGAAGAAGGCGTAGATCTCGTCCTCGTTGGTGTCGCGGAAGGTG 1682
Qy 2017 GCGCTGGTCATGCTCTCGCGGCGCTTCAGGCTGATCTCGGCGTTGGTCCACTTCACGATC 2076
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1683 GCGCTGGTCATGCTCTCGCGGCGCTTCAGGCTGATCTCGGCGTTGGTCCACTTCACGATC 1742
Qy 2077 TCGCTGATGATCTCGTCGGTGAAGAACAGCTTGAAGCACAGCAGGGGGTCGTAGATGTTG 2136
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1743 TCGCTGATGATCTCGTCGGTGAAGAACAGCTTGAAGCACAGCAGGGGGTCGTAGATGTTG 1802
Qy 2137 CGGCACATGCGGGTGGGGCCGCGCTGGCTGCGCACGATGTTCAGGGCGCTCACGCGGCTG 2196
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1803 CGGCACATGCGGGTGGGGCCGCGCTGGCTGCGCACGATGTTCAGGGCGCTCACGCGGCTG 1862
Qy 2197 CGGCGGGTGGGCTTGCTGGTGCTCCAGCAGTGCTTGTTCTTGCCGCGGATGGTGCGCTGG 2256
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1863 CGGCGGGTGGGCTTGCTGGTGCTCCAGCAGTGCTTGTTCTTGCCGCGGATGGTGCGCTGG 1922
Qy 2257 GGCAGGGTCAGGATGCGGTTGCTGGCCAGGCTGCTGCCGGGCTGCTCGATCACGTTCTGC 2316
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1923 GGCAGGGTCAGGATGCGGTTGCTGGCCAGGCTGCTGCCGGGCTGCTCGATCACGTTCTGC 1982
Qy 2317 TCGTCCAGGATCTCGCTGCCGCTGCTGGTGGGCTGCACCTCGTGCACCTCGTCGATGAAG 2376
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1983 TCGTCCAGGATCTCGCTGCCGCTGCTGGTGGGCTGCACCTCGTGCACCTCGTCGATGAAG 2042
Qy 2377 GCCTCCTCGGTGTCGCTCTGCACGTCGTCCTCGCTCACGTGGTCGCTCACCTCGCTGTCG 2436
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2043 GCCTCCTCGGTGTCGCTCTGCACGTCGTCCTCGCTCACGTGGTCGCTCACCTCGCTGTCG 2102
Qy 2437 CTGTCCTCGCCCACCAGCTCGTCGTCGCTCTGCAGCAGGGCGCTCAGGATGTGCTCGTCG 2496
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2103 CTGTCCTCGCCCACCAGCTCGTCGTCGCTCTGCAGCAGGGCGCTCAGGATGTGCTCGTCG 2162
Qy 2497 TCCAGGCTGCTGCCATCTTCTGTTTTGATTTTCTTAGGTTTGCCCATGGTGCCAAAGTTG 2556
|||||||||||||| || | | || || |||||||||| ||| |||
Db 2163 TCCAGGCTGCTGCCGTCCAACTTGACCCTCTTGGCAGCAGGGCCCATGGTGGCAAGCTTG 2222
Qy 2557 AGC 2559
| |
Db 2223 ATC 2225
APPENDIX C
Qian et al. with SEQ ID NO: 7
Query Match 96.7%; Score 1781.6; DB 1; Length 1815;
Best Local Similarity 99.0%;
Matches 1793; Conservative 0; Mismatches 19; Indels 0; Gaps 0;
Qy 1 ATGGGCAAACCTAAGAAAATCAAAACAGAAGATGGCAGCAGCCTGGACGACGAGCACATC 60
|||||| || || | | || |||||||||||||||||||||||||||
Db 1 ATGGGCCCTGCTGCCAAGAGGGTCAAGTTGGACGGCAGCAGCCTGGACGACGAGCACATC 60
Qy 61 CTGAGCGCCCTGCTGCAGAGCGACGACGAGCTGGTGGGCGAGGACAGCGACAGCGAGGTG 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 CTGAGCGCCCTGCTGCAGAGCGACGACGAGCTGGTGGGCGAGGACAGCGACAGCGAGGTG 120
Qy 121 AGCGACCACGTGAGCGAGGACGACGTGCAGAGCGACACCGAGGAGGCCTTCATCGACGAG 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 AGCGACCACGTGAGCGAGGACGACGTGCAGAGCGACACCGAGGAGGCCTTCATCGACGAG 180
Qy 181 GTGCACGAGGTGCAGCCCACCAGCAGCGGCAGCGAGATCCTGGACGAGCAGAACGTGATC 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 GTGCACGAGGTGCAGCCCACCAGCAGCGGCAGCGAGATCCTGGACGAGCAGAACGTGATC 240
Qy 241 GAGCAGCCCGGCAGCAGCCTGGCCAGCAACCGCATCCTGACCCTGCCCCAGCGCACCATC 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 GAGCAGCCCGGCAGCAGCCTGGCCAGCAACCGCATCCTGACCCTGCCCCAGCGCACCATC 300
Qy 301 CGCGGCAAGAACAAGCACTGCTGGAGCACCAGCAAGCCCACCCGCCGCAGCCGCGTGAGC 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 CGCGGCAAGAACAAGCACTGCTGGAGCACCAGCAAGCCCACCCGCCGCAGCCGCGTGAGC 360
Qy 361 GCCCTGAACATCGTGCGCAGCCAGCGCGGCCCCACCCGCATGTGCCGCAACATCTACGAC 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 GCCCTGAACATCGTGCGCAGCCAGCGCGGCCCCACCCGCATGTGCCGCAACATCTACGAC 420
Qy 421 CCCCTGCTGTGCTTCAAGCTGTTCTTCACCGACGAGATCATCAGCGAGATCGTGAAGTGG 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 CCCCTGCTGTGCTTCAAGCTGTTCTTCACCGACGAGATCATCAGCGAGATCGTGAAGTGG 480
Qy 481 ACCAACGCCGAGATCAGCCTGAAGCGCCGCGAGAGCATGACCAGCGCCACCTTCCGCGAC 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 ACCAACGCCGAGATCAGCCTGAAGCGCCGCGAGAGCATGACCAGCGCCACCTTCCGCGAC 540
Qy 541 ACCAACGAGGACGAGATCTACGCCTTCTTCGGCATCCTGGTGATGACCGCCGTGCGCAAG 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 ACCAACGAGGACGAGATCTACGCCTTCTTCGGCATCCTGGTGATGACCGCCGTGCGCAAG 600
Qy 601 GACAACCACATGAGCACCGACGACCTGTTCGACCGCAGCCTGAGCATGGTGTACGTGAGC 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 GACAACCACATGAGCACCGACGACCTGTTCGACCGCAGCCTGAGCATGGTGTACGTGAGC 660
Qy 661 GTGATGAGCCGCGACCGCTTCGACTTCCTGATCCGCTGCCTGCGCATGGACGACAAGAGC 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 GTGATGAGCCGCGACCGCTTCGACTTCCTGATCCGCTGCCTGCGCATGGACGACAAGAGC 720
Qy 721 ATCCGCCCCACCCTGCGCGAGAACGACGTGTTCACCCCCGTGCGCAAGATCTGGGACCTG 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 721 ATCCGCCCCACCCTGCGCGAGAACGACGTGTTCACCCCCGTGCGCAAGATCTGGGACCTG 780
Qy 781 TTCATCCACCAGTGCATCCAGAACTACACCCCCGGCGCCCACCTGACCATCGACGAGCAG 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 781 TTCATCCACCAGTGCATCCAGAACTACACCCCCGGCGCCCACCTGACCATCGACGAGCAG 840
Qy 841 CTGCTGGGCTTCCGCGGCCGCTGCCCCTTCCGCGTGTACATCCCCAACAAGCCCAGCAAG 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 841 CTGCTGGGCTTCCGCGGCCGCTGCCCCTTCCGCGTGTACATCCCCAACAAGCCCAGCAAG 900
Qy 901 TACGGCATCAAGATCCTGATGATGTGCGACAGCGGCACCAAGTACATGATCAACGGCATG 960
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 901 TACGGCATCAAGATCCTGATGATGTGCGACAGCGGCACCAAGTACATGATCAACGGCATG 960
Qy 961 CCCTACCTGGGCCGCGGCACCCAGACCAACGGCGTGCCCCTGGGCGAGTACTACGTGAAG 1020
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 961 CCCTACCTGGGCCGCGGCACCCAGACCAACGGCGTGCCCCTGGGCGAGTACTACGTGAAG 1020
Qy 1021 GAGCTGAGCAAGCCCGTGCACGGCAGCTGCCGCAACATCACCTGCGACAACTGGTTCACC 1080
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1021 GAGCTGAGCAAGCCCGTGCACGGCAGCTGCCGCAACATCACCTGCGACAACTGGTTCACC 1080
Qy 1081 AGCATCCCCCTGGCCAAGAACCTGCTGCAGGAGCCCTACAAGCTGACCATCGTGGGCACC 1140
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1081 AGCATCCCCCTGGCCAAGAACCTGCTGCAGGAGCCCTACAAGCTGACCATCGTGGGCACC 1140
Qy 1141 GTGCGCAGCAACAAGCGCGAGATCCCCGAGGTGCTGAAGAACAGCCGCAGCCGCCCCGTG 1200
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1141 GTGCGCAGCAACAAGCGCGAGATCCCCGAGGTGCTGAAGAACAGCCGCAGCCGCCCCGTG 1200
Qy 1201 GGCACCAGCATGTTCTGCTTCGACGGCCCCCTGACCCTGGTGAGCTACAAGCCCAAGCCC 1260
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1201 GGCACCAGCATGTTCTGCTTCGACGGCCCCCTGACCCTGGTGAGCTACAAGCCCAAGCCC 1260
Qy 1261 GCCAAGATGGTGTACCTGCTGAGCAGCTGCGACGAGGACGCCAGCATCAACGAGAGCACC 1320
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1261 GCCAAGATGGTGTACCTGCTGAGCAGCTGCGACGAGGACGCCAGCATCAACGAGAGCACC 1320
Qy 1321 GGCAAGCCCCAGATGGTGATGTACTACAACCAGACCAAGGGCGGCGTGGACACCCTGGAC 1380
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1321 GGCAAGCCCCAGATGGTGATGTACTACAACCAGACCAAGGGCGGCGTGGACACCCTGGAC 1380
Qy 1381 CAGATGTGCAGCGTGATGACCTGCAGCCGCAAGACCAACCGCTGGCCCATGGCCCTGCTG 1440
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1381 CAGATGTGCAGCGTGATGACCTGCAGCCGCAAGACCAACCGCTGGCCCATGGCCCTGCTG 1440
Qy 1441 TACGGCATGATCAACATCGCCTGCATCAACAGCTTCATCATCTACAGCCACAACGTGAGC 1500
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1441 TACGGCATGATCAACATCGCCTGCATCAACAGCTTCATCATCTACAGCCACAACGTGAGC 1500
Qy 1501 AGCAAGGGCGAGAAGGTGCAGAGCCGCAAGAAGTTCATGCGCAACCTGTACATGGGCCTG 1560
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1501 AGCAAGGGCGAGAAGGTGCAGAGCCGCAAGAAGTTCATGCGCAACCTGTACATGGGCCTG 1560
Qy 1561 ACCAGCAGCTTCATGCGCAAGCGCCTGGAGGCCCCCACCCTGAAGCGCTACCTGCGCGAC 1620
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1561 ACCAGCAGCTTCATGCGCAAGCGCCTGGAGGCCCCCACCCTGAAGCGCTACCTGCGCGAC 1620
Qy 1621 AACATCAGCAACATCCTGCCCAAGGAGGTGCCCGGCACCAGCGACGACAGCACCGAGGAG 1680
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1621 AACATCAGCAACATCCTGCCCAAGGAGGTGCCCGGCACCAGCGACGACAGCACCGAGGAG 1680
Qy 1681 CCCGTGATGAAGAAGCGCACCTACTGCACCTACTGCCCCAGCAAGATCCGCCGCAAGGCC 1740
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1681 CCCGTGATGAAGAAGCGCACCTACTGCACCTACTGCCCCAGCAAGATCCGCCGCAAGGCC 1740
Qy 1741 AGCGCCAGCTGCAAGAAGTGCAAGAAGGTGATCTGCCGCGAGCACAACATCGACATGTGC 1800
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1741 AGCGCCAGCTGCAAGAAGTGCAAGAAGGTGATCTGCCGCGAGCACAACATCGACATGTGC 1800
Qy 1801 CAGAGCTGCTTC 1812
||||||||||||
Db 1801 CAGAGCTGCTTC 1812
Qian et al. with SEQ ID NO: 35
Query Match 100.0%; Score 27; Length 27;
Best Local Similarity 100.0%;
Matches 27; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 CCTGCTGCCAAGAGGGTCAAGTTGGAC 27
|||||||||||||||||||||||||||
Db 1 CCTGCTGCCAAGAGGGTCAAGTTGGAC 27