pDRESS_mTurquoise2_spatial-linker-P2A_mScarlet3

Plasmid Information

Purpose

Produces an mTurquoise-antiFRET-mScarlet3 fusion protein in bacteria and in mamallian cells it produces 1:1 unfused mTurquoise2 and mScarlet3 FPs for ratiometric analysis

Depositor

Dorus Gadella

Article

Gadella TWJ et al Nat Methods. 2023 Apr;20(4):541-545. doi

Insert

mTurquoise2_spatial-linker-P2A_mScarlet3

Sequence Information

Length

6,559 bp

Topology

circular

Molecule Type

other DNA

View on AddGene.org

Features

3'UTR 1

Label	Gene	Product	Position	Strand
polyhedrin 3' UTR	—	—	2485..2858	Fwd
DNA Sequence AACACGATACATTGTTATTAGTACATTTATTAAGCGCTAGATTCTGTGCGTTGTTGATTTACAGACAATTGTTGTACGTATTTTAATAATTCATTAAATTTATAATCTTTAGGGTGGTATGTTAGAGCGAAAATCAAATGATTTTCAGCGTCTTTATATCTGAATTTAAATATTAAATCCTCAATAGATTTGTAAAATAGGTTTCGATTAGTTTCAAACAAGGGTTGTTTTTCCGAACCGATGGCTGGACTATCTAATGGATTTTCGCTCAACGCCACAAAACTTGCCAAATCTTGTAGCAGCAATCTAGCTTTGTCGATATTCGTTTGTGTTTTGTTTTGTAATAAAGGTTCGACGTCGTTCAAAATATTAT

CDS 9

Label	Gene	Product	Position	Strand
6xHis	—	6xHis affinity tag	354..372	Fwd
DNA Sequence CATCATCACCACCATCAC Protein Sequence HHHHHH
thrombin site	—	thrombin recognition and cleavage site	381..399	Fwd
DNA Sequence CTGGTGCCGCGCGGCAGC Protein Sequence LVPRGS
mTurquoise2	—	enhanced monomeric variant of CFP (Goed…	423..1140	Fwd
DNA Sequence ATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCCTGTCCTGGGGCGTGCAGTGCTTCGCCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACTTTAGCGACAACGTCTATATCACCGCCGACAAGCAGAAGAACGGCATCAAGGCCAACTTCAAGATCCGCCACAACATCGAGGACGGCGGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCACCCAGTCCAAGCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTCTACAAG Protein Sequence MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK
P2A	—	2A peptide from porcine teschovirus-1 p…	1260..1317	Fwd
DNA Sequence GCTACTAACTTCAGCCTGCTGAAGCAGGCTGGAGACGTGGAGGAGAACCCTGGACCT Protein Sequence ATNFSLLKQAGDVEENPGP
mScarlet3	—	bright cysteine-free monomeric red fluo…	1341..2028	Fwd
DNA Sequence ATGGATAGCACCGAGGCAGTGATCAAGGAGTTCATGCGGTTCAAGGTGCACATGGAGGGCTCCATGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCTACGAGGGCACCCAGACCGCCAAGCTGAGGGTGACCAAGGGTGGCCCCCTGCCCTTCTCCTGGGACATCCTGTCCCCTCAGTTCATGTACGGCTCCAGGGCCTTCACGAAGCACCCCGCCGACATCCCCGACTACTGGAAGCAGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGAGGACGGCGGCGCCGTGTCCGTGGCCCAGGACACCTCCCTGGAGGACGGCACCCTGATCTACAAGGTGAAGCTCCGCGGCACCAACTTCCCTCCTGACGGCCCCGTAATGCAGAAGAAGACAATGGGCTGGGAAGCATCCACCGAGCGGTTGTACCCCGAGGACGTCGTGCTGAAGGGCGACATTAAGATGGCCCTGCGCCTGAAGGACGGCGGCCGCTACCTGGCGGACTTCAAGACCACCTACAGGGCCAAGAAGCCCGTGCAGATGCCCGGCGCCTTCAACATCGACCGCAAGTTGGACATCACATCCCACAACGAGGACTACACCGTGGTGGAACAGTACGAACGCTCCGTGGCCCGCCACTCCACCGGCGGCTCCGGTGGCTCC Protein Sequence MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAKLRVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYWKQSFPEGFKWERVMNFEDGGAVSVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKMALRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHSTGGSGGS
ORF1629	—	baculovirus capsid-associated protein	2514..3103	Rev
DNA Sequence AATTCCAAAAAGCTCAACTACAAATTTGATCGCGGACGTGTTAGCCGACACAATTAATAGGCGTCGTGTGGCTATGGCAAAATCGTCTTCGGAAGCAACTTCTAACGACGAGGGTTGGGACGACGACGATAATCGGCCTAATAAAGCTAACACGCCCGATGTTAAATATGTCCAAGCTCTATTTAACGTGTTTACGTCGAGTCAATTGTATACTAACGACAGTGATGAAAGAAATACAAAAGCGCATAATATTTTGAACGACGTCGAACCTTTATTACAAAACAAAACACAAACGAATATCGACAAAGCTAGATTGCTGCTACAAGATTTGGCAAGTTTTGTGGCGTTGAGCGAAAATCCATTAGATAGTCCAGCCATCGGTTCGGAAAAACAACCCTTGTTTGAAACTAATCGAAACCTATTTTACAAATCTATTGAGGATTTAATATTTAAATTCAGATATAAAGACGCTGAAAATCATTTGATTTTCGCTCTAACATACCACCCTAAAGATTATAAATTTAATGAATTATTAAAATACGTACAACAATTGTCTGTAAATCAACAACGCACAGAATCTAGCGCTTAA Protein Sequence IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNRPNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKARLLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIFALTYHPKDYKFNELLKYVQQLSVNQQRTESSA
KanR	aph(3')-Ia	aminoglycoside phosphotransferase	3912..4728	Rev
DNA Sequence ATGAGCCATATTCAACGGGAAACGTCTTGCTCGAGGCCGCGATTAAATTCCAACATGGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGATTGTATGGGAAGCCCGATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGGCTAAACTGGCTGACGGAATTTATGCCTCTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCAGGGAAAACAGCATTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCATTCGATTCCTGTTTGTAATTGTCCTTTTAACGGCGATCGCGTATTTCGTCTCGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTGGTGCGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAGCTTTTGCCATTCTCACCGGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACGAGTCGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTTTTCAAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCACTTGATGCTCGATGAGTTTTTCTAA Protein Sequence MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF
lef2	—	baculovirus late expression factor 2	4970..5095	Fwd
DNA Sequence TGCATCTTTTAATCAAATCCCAAGATGTGTATAAACCACCAAACTGCCAAAAAATGAAAACTGTCGACAAGCTCTGTCCGTTTGCTGGCAACTGCAAGGGTCTCAATCCTATTTGTAATTATTGA Protein Sequence HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY
ORF603	—	baculovirus ORF603 protein	5132..5738	Rev
DNA Sequence ATGGCTGTTATTTTTAACAACAAACAACTGCTCGCAGACAATAGTATAGAAAAGGGAGGTGAACTGTTTTTGTTTAACGGTTCGTACAACATTTTGGAAAGTTATGTTAATCCGGTGCTGCTAAAAAATGGTGTAATTGAACTAGAAGAAGCTGCGTACTATGCCGGCAACATATTGTATAAAACCGACGATCCCAAATTCATTGATTATATAAATTTAATAATTAAAGCAACACACTCCGAAGAACTACCAGAAAATAGCACTGTTGTAAATTACAGAAAAACTATGCGCAGCGGTACTATACACCCCATTAAAAAAGACATATATATTTATGACAACAAAAAATTTACTCTATACGATAGATACATATATGGATACGATAATAACTATGTTAATTTTTATGAAGAGAAAAATGAAAAAGAGAAGGAATACGAAGAAGAAGACGACAAGGCGTCTAGTTTATGTGAAAATAAAATTATATTGTCGCAAATTAACTGTGAATCATTTGAAAATGATTTTAAATATTACCTCAGCGATTATAACTACGCGTTTTCAATTATAGATAATACTACAAATGTTCTTGTTGCGTTTGGTTTGTATCGTTAA Protein Sequence MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGVIELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPIKKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQINCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR

enhancer 1

Label	Gene	Product	Position	Strand
CMV enhancer	—	—	5865..6169	Fwd
DNA Sequence CGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTCCGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTACGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATG

polyA_signal 1

Label	Gene	Product	Position	Strand
beta-globin poly(A) signal	—	—	2194..2250	Fwd
DNA Sequence AATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCA

promoter 2

Label	Gene	Product	Position	Strand
rhaB promoter	—	—	161..280	Fwd
DNA Sequence CACCACAATTCAGCAAATTGTGAACATCATCACGTTCATCTTTCCCTGGTTGCCAAAGGCCCATTTTCCTGTCAGTAACGAGAAGGTCGCGAATTCAGGCGCTTTTTAGACTGGTCGTA
CMV promoter	—	—	6170..6373	Fwd
DNA Sequence TGATGCGGTTTTGGCAGTACACCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAATAACCCCGCCCCGTTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGACGT

protein_bind 1

Label	Gene	Product	Position	Strand
loxP	—	—	3111..3145	Rev
DNA Sequence ATAACTTCGTATAATGTATGCTATACGAAGTTAT

rep_origin 1

Label	Gene	Product	Position	Strand
ori	—	—	3312..3900	Rev
DNA Sequence TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGCGGAGCCTATGGAAA

terminator 1

Label	Gene	Product	Position	Strand
T7 terminator	—	—	2337..2385	Fwd
DNA Sequence CTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG