Dox-Akt — PlasmidDB

Plasmid Information

Purpose

PiggyBac vector backbone encoding doxycycline-inducible expression of membrane-localized Akt1 (without pleckstrin homology domain)

Depositor

Jared Toettcher

Article

Underhill et al Development. 2023 Aug 15;150(16):dev2016

Insert

rac protein kinase-alpha (AKT1 Human)

Sequence Information

Length

9,825 bp

Topology

circular

Molecule Type

other DNA

View on AddGene.org

Features

CDS 10

Label	Gene	Product	Position	Strand
FusionRed	—	monomeric red fluorescent protein optim…	74..770	Fwd
DNA Sequence ATGGTGAGCGAGCTGATTAAGGAGAACATGCCCATGAAGCTGTACATGGAGGGCACCGTGAACAACCACCACTTCAAGTGCACATCCGAGGGCGAAGGCAAGCCCTACGAGGGCACCCAGACCATGAGAATCAAGGTCGTCGAGGGCGGCCCTCTCCCCTTCGCCTTCGACATCCTGGCTACCAGCTTCATGTACGGCAGCAGAACCTTCATCAAGCACCCTCCGGGCATCCCCGACTTCTTTAAGCAGTCCTTCCCTGAGGGCTTCACATGGGAGAGAGTCACCACATACGAAGACGGGGGCGTGCTGACCGCTACCCAGGACACCAGCCTCCAGGACGGCTGCCTCATCTACAACGTCAAGGTTAGAGGGGTGAACTTCCCAGCCAACGGCCCTGTGATGCAGAAGAAAACACTCGGCTGGGAGGCCTCCACCGAGACGATGTACCCCGCTGACGGCGGCCTGGAAGGCGCATGTGACATGGCCCTGAAGCTCGTGGGCGGGGGCCACCTGATCTGCAACCTTGAGACCACATACAGATCCAAGAAACCCGCTACGAACCTCAAGATGCCCGGCGTCTACAACGTGGACCACAGACTGGAAAGAATCAAGGAGGCCGACGATGAGACCTACGTCGAGCAGCACGAGGTGGCTGTGGCCAGATACTCTACTGGTGGCGCTGGTGATGGAGGTAAA Protein Sequence MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKVVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLTATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMALKLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARYSTGGAGDGGK
P2A	—	2A peptide from porcine teschovirus-1 p…	779..836	Fwd
DNA Sequence GCTACTAACTTCAGCCTGCTGAAGCAGGCTGGAGACGTGGAGGAGAACCCTGGACCT Protein Sequence ATNFSLLKQAGDVEENPGP
Tet-On(R) 3G	—	modified rtTA protein that binds tightl…	836..1583	Fwd
DNA Sequence ATGAGCCGGCTGGACAAGAGCAAAGTGATCAACAGCGCCCTGGAACTGCTGAACGGCGTGGGCATCGAGGGCCTGACCACCCGGAAGCTGGCCCAGAAACTGGGCGTGGAACAGCCCACCCTGTACTGGCACGTGAAGAACAAGCGGGCCCTGCTGGACGCCCTGCCCATCGAGATGCTGGACCGGCACCACACCCACAGCTGCCCTCTGGAAGGCGAGAGCTGGCAGGACTTCCTGCGGAACAACGCCAAGAGCTACAGATGCGCCCTGCTGAGCCACCGGGACGGCGCCAAAGTGCACCTGGGCACCAGACCCACCGAGAAGCAGTACGAGACACTGGAAAACCAGCTGGCCTTCCTGTGCCAGCAGGGCTTCAGCCTGGAAAACGCCCTGTACGCCCTGAGCGCCGTGGGCCACTTCACCCTGGGCTGCGTGCTGGAAGAACAGGAACACCAGGTCGCCAAAGAGGAAAGAGAGACACCCACCACCGACAGCATGCCCCCCCTGCTGAAGCAGGCCATCGAGCTGTTCGACAGACAGGGCGCCGAGCCCGCCTTCCTGTTCGGCCTGGAACTGATCATCTGCGGCCTCGAGAAGCAGCTGAAGTGCGAGAGCGGCGGACCCACCGACGCCCTGGACGACTTCGACCTGGACATGCTGCCCGCCGATGCCCTGGATGATTTTGATCTGGATATGCTCCCTGCTGACGCTCTCGATGACTTCGATCTCGATATGCTGCCAGGCTAG Protein Sequence MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHVKNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTRPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTTDSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPADALDDFDLDMLPADALDDFDLDMLPG
Factor Xa site	—	Factor Xa recognition and cleavage site	2158..2170	Rev
DNA Sequence ATTGAGGGCCGA Protein Sequence IEGR
AmpR	bla	beta-lactamase	3062..3923	Fwd
DNA Sequence ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA Protein Sequence MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
myr	—	N-myristoylation signal from Src kinase…	5987..6008	Fwd
DNA Sequence ATGGGTTCCTCCAAATCCAAG Protein Sequence MGSSKSK
TagBFP	—	monomeric blue fluorescent protein	6035..6734	Fwd
DNA Sequence ATGAGCGAGCTGATTAAGGAGAACATGCACATGAAGCTGTACATGGAGGGCACCGTGGACAACCATCACTTCAAGTGCACATCCGAGGGCGAAGGCAAGCCCTACGAGGGCACCCAGACCATGAGAATCAAGGTGGTCGAGGGCGGCCCTCTCCCCTTCGCCTTCGACATCCTGGCTACTAGCTTCCTCTACGGCAGCAAGACCTTCATCAACCACACCCAGGGCATCCCCGACTTCTTCAAGCAGTCCTTCCCTGAGGGCTTCACATGGGAGAGAGTCACCACATACGAAGACGGGGGCGTGCTGACCGCTACCCAGGACACCAGCCTCCAGGACGGCTGCCTCATCTACAACGTCAAGATCAGAGGGGTGAACTTCACATCCAACGGCCCTGTGATGCAGAAGAAAACACTCGGCTGGGAGGCCTTCACCGAGACGCTGTACCCCGCTGACGGCGGCCTGGAAGGCAGAAACGACATGGCCCTGAAGCTCGTGGGCGGGAGCCATCTGATCGCAAACATCAAGACCACATATAGATCCAAGAAACCCGCTAAGAACCTCAAGATGCCTGGCGTCTACTATGTGGACTACAGACTGGAAAGAATCAAGGAGGCCAACAACGAGACCTACGTCGAGCAGCACGAGGTGGCAGTGGCCAGATACTGCGACCTCCCTAGCAAACTGGGGCACAAGCTTAAT Protein Sequence MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKLVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCDLPSKLGHKLN
HA	—	HA (human influenza hemagglutinin) epit…	7817..7844	Fwd
DNA Sequence TATCCTTACGACGTCCCCGACTACGCC Protein Sequence YPYDVPDYA
HA	—	HA (human influenza hemagglutinin) epit…	7823..7850	Fwd
DNA Sequence TACGACGTCCCCGACTACGCCAGTCTG Protein Sequence YDVPDYASL
Factor Xa site	—	Factor Xa recognition and cleavage site	8440..8452	Rev
DNA Sequence ATTGAGGGCCGA Protein Sequence IEGR

intron 1

Label	Gene	Product	Position	Strand
EF-1-alpha intron A	—	—	8886..9825	Fwd
DNA Sequence GTAAGTGCCGTGTGTGGTTCCCGCGGGCCTGGCCTCTTTACGGGTTATGGCCCTTGCGTGCCTTGAATTACTTCCACCTGGCTGCAGTACGTGATTCTTGATCCCGAGCTTCGGGTTGGAAGTGGGTGGGAGAGTTCGAGGCCTTGCGCTTAAGGAGCCCCTTCGCCTCGTGCTTGAGTTGAGGCCTGGCCTGGGCGCTGGGGCCGCCGCGTGCGAATCTGGTGGCACCTTCGCGCCTGTCTCGCTGCTTTCGATAAGTCTCTAGCCATTTAAAATTTTTGATGACCTGCTGCGACGCTTTTTTTCTGGCAAGATAGTCTTGTAAATGCGGGCCAAGATCTGCACACTGGTATTTCGGTTTTTGGGGCCGCGGGCGGCGACGGGGCCCGTGCGTCCCAGCGCACATGTTCGGCGAGGCGGGGCCTGCGAGCGCGGCCACCGAGAATCGGACGGGGGTAGTCTCAAGCTGGCCGGCCTGCTCTGGTGCCTGGCCTCGCGCCGCCGTGTATCGCCCCGCCCTGGGCGGCAAGGCTGGCCCGGTCGGCACCAGTTGCGTGAGCGGAAAGATGGCCGCTTCCCGGCCCTGCTGCAGGGAGCTCAAAATGGAGGACGCGGCGCTCGGGAGAGCGGGCGGGTGAGTCACCCACACAAAGGAAAAGGGCCTTTCCGTCCTCAGCCGTCGCTTCATGTGACTCCACGGAGTACCGGGCGCCGTCCAGGCACCTCGATTAGTTCTCGAGCTTTTGGAGTACGTCGTCTTTAGGTTGGGGGGAGGGGTTTTATGCGATGGAGTTTCCCCACACTGAGTGGGTGGAGACTGAAGTTAGGCCAGCTTGGCACTTGATGTAATTCTCCTTGGAATTTGCCCTTTTTAAGTTTGGATCTTGGTTCATTCTCAAGCCTCAGACAGTGGTTCAAAGTTTTTTTCTTCCATTTCAG

misc_feature 2

Label	Gene	Product	Position	Strand
WPRE	—	—	1687..2276	Fwd
DNA Sequence AATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGC
WPRE	—	—	7969..8558	Fwd
DNA Sequence AATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGC

promoter 3

Label	Gene	Product	Position	Strand
AmpR promoter	bla	—	2957..3062	Fwd
DNA Sequence CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
lac promoter	—	—	5005..5036	Fwd
DNA Sequence TTTACACTTTATGCTTCCGGCTCGTATGTTG
TRE3G promoter	—	—	5533..5912	Fwd
DNA Sequence GGCTTTACTCCCTATCAGTGATAGAGAACGTATGAAGAGTTTACTCCCTATCAGTGATAGAGAACGTATGCAGACTTTACTCCCTATCAGTGATAGAGAACGTATAAGGAGTTTACTCCCTATCAGTGATAGAGAACGTATGACCAGTTTACTCCCTATCAGTGATAGAGAACGTATCTACAGTTTACTCCCTATCAGTGATAGAGAACGTATATCCAGTTTACTCCCTATCAGTGATAGAGAACGTATAAGCTTTAGGCGTGTACGGTGGGCGCCTATAAAAGCAGAGCTCGTTTAGTGAACCGTCAGATCGCCTGGAGCAATTCCACAACACTTTTGTCTTATACCAACTTTCCGTACCACTTCCTACCCTCGTAAA

protein_bind 12

Label	Gene	Product	Position	Strand
CAP binding site	—	—	4969..4991	Fwd
DNA Sequence TAATGTGAGTTAGCTCACTCAT
lac operator	—	—	5043..5060	Fwd
DNA Sequence TTGTGAGCGGATAACAA
tet operator	tetO	—	5541..5560	Fwd
DNA Sequence TCCCTATCAGTGATAGAGA
tet operator	tetO	—	5577..5596	Fwd
DNA Sequence TCCCTATCAGTGATAGAGA
tet operator	tetO	—	5613..5632	Fwd
DNA Sequence TCCCTATCAGTGATAGAGA
tet operator	tetO	—	5649..5668	Fwd
DNA Sequence TCCCTATCAGTGATAGAGA
tet operator	tetO	—	5685..5704	Fwd
DNA Sequence TCCCTATCAGTGATAGAGA
tet operator	tetO	—	5721..5740	Fwd
DNA Sequence TCCCTATCAGTGATAGAGA
tet operator	tetO	—	5757..5776	Fwd
DNA Sequence TCCCTATCAGTGATAGAGA
attB1	mutant version of attB	—	5957..5982	Fwd
DNA Sequence ACAAGTTTGTACAAAAAAGCAGGCT
tet operator	tetO	—	8802..8821	Fwd
DNA Sequence TCCCTATCAGTGATAGAGA
tet operator	tetO	—	8823..8842	Fwd
DNA Sequence TCCCTATCAGTGATAGAGA

rep_origin 1

Label	Gene	Product	Position	Strand
ori	—	—	4093..4682	Fwd
DNA Sequence TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA