US 12,385,069 B2
Compositions and methods for studying the Tat gene
Martin R. Schiller, Henderson, NV (US); and Ronald Benjamin, Henderson, NV (US)
Assigned to The Board Of Regents Of The Nevada System Of Higher Education On Behalf Of The University of Nevada, Las Vegas, NV (US)
Appl. No. 16/318,965
Filed by The Board Of Regents Of The Nevada System Of Higher Education On Behalf Of The University of Nevada, Las Vegas, Las Vegas, NV (US)
PCT Filed Jul. 14, 2017, PCT No. PCT/US2017/042179
§ 371(c)(1), (2) Date Jan. 18, 2019,
PCT Pub. No. WO2018/017419, PCT Pub. Date Jan. 25, 2018.
Claims priority of provisional application 62/365,537, filed on Jul. 22, 2016.
Prior Publication US 2020/0017883 A1, Jan. 16, 2020
Int. Cl. C40B 20/04 (2006.01); C07K 14/005 (2006.01); C12N 15/10 (2006.01); C12N 15/11 (2006.01); C12N 15/113 (2010.01); C12N 15/65 (2006.01); C12N 15/90 (2006.01); C40B 40/08 (2006.01)
CPC C12N 15/907 (2013.01) [C07K 14/005 (2013.01); C12N 15/1065 (2013.01); C12N 15/1086 (2013.01); C12N 15/111 (2013.01); C12N 15/113 (2013.01); C12N 15/65 (2013.01); C40B 20/04 (2013.01); C40B 40/08 (2013.01); C12N 2740/16322 (2013.01); C12N 2750/14122 (2013.01)] 1 Claim
 
1. A vector comprising a double-stranded nucleic acid construct, wherein the double-stranded nucleic acid construct comprises a first strand and a second strand, wherein the first strand comprises from 5′ to 3′, a left arm of a AAVS1 locus sequence;
a nucleic acid sequence encoding puromycin N-acetyl-transferase; a CMV promoter; a tat cDNA coding sequence; a 3′ UTR, wherein the 3′ UTR comprises a nucleic acid sequence complementary to a synthetic poly(A) signal of the 3′UTR of the second strand, a first barcode sequence complementary to a second barcode sequence of the second strand, and a polySV40(A) sequence; a nucleic acid sequence complementary to a GFP coding sequence of the second strand, a sequence complementary to a LTR promoter of the second strand; a sequence complementary to the right arm of the AAVS1 locus sequence of the second strand; and a functional sequence;
wherein the second strand comprises from 5′ to 3′, a right arm of a AAVS1 locus sequence; a LTR promoter; a GFP coding sequence; a 3′ UTR, wherein the 3′UTR comprises a nucleic acid sequence complementary to the polySV40(A) sequence of the 3′ UTR of the first strand, the second barcode sequence complementary to the first barcode sequence of the first strand, and a synthetic poly(A) signal; a nucleic acid sequence complementary to the tat cDNA coding sequence and the CMV promoter of the first strand; a nucleic acid sequence encoding puromycin N-acetyl-transferase; a nucleic acid sequence complementary to the left arm of the AAVS1 locus sequence of the first strand; and a functional sequence.