pDRESS_mTurquoise2_spatial-linker-P2A_mScarlet-I3

Plasmid Information

Purpose

Produces an mTurquoise-antiFRET-mScarlet-I3 fusion protein in bacteria and in mamallian cells it produces 1:1 unfused mTurquoise2 and mScarlet-I3 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_mScarlet-I3

Sequence Information

Length

6,556 bp

Topology

circular

Molecule Type

other DNA

View on AddGene.org

Features

3'UTR 1

Label	Gene	Product	Position	Strand
polyhedrin 3' UTR	—	—	4095..4468	Fwd
DNA Sequence AACACGATACATTGTTATTAGTACATTTATTAAGCGCTAGATTCTGTGCGTTGTTGATTTACAGACAATTGTTGTACGTATTTTAATAATTCATTAAATTTATAATCTTTAGGGTGGTATGTTAGAGCGAAAATCAAATGATTTTCAGCGTCTTTATATCTGAATTTAAATATTAAATCCTCAATAGATTTGTAAAATAGGTTTCGATTAGTTTCAAACAAGGGTTGTTTTTCCGAACCGATGGCTGGACTATCTAATGGATTTTCGCTCAACGCCACAAAACTTGCCAAATCTTGTAGCAGCAATCTAGCTTTGTCGATATTCGTTTGTGTTTTGTTTTGTAATAAAGGTTCGACGTCGTTCAAAATATTAT

CDS 9

Label	Gene	Product	Position	Strand
lef2	—	baculovirus late expression factor 2	24..149	Fwd
DNA Sequence TGCATCTTTTAATCAAATCCCAAGATGTGTATAAACCACCAAACTGCCAAAAAATGAAAACTGTCGACAAGCTCTGTCCGTTTGCTGGCAACTGCAAGGGTCTCAATCCTATTTGTAATTATTGA Protein Sequence HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY
ORF603	—	baculovirus ORF603 protein	186..792	Rev
DNA Sequence ATGGCTGTTATTTTTAACAACAAACAACTGCTCGCAGACAATAGTATAGAAAAGGGAGGTGAACTGTTTTTGTTTAACGGTTCGTACAACATTTTGGAAAGTTATGTTAATCCGGTGCTGCTAAAAAATGGTGTAATTGAACTAGAAGAAGCTGCGTACTATGCCGGCAACATATTGTATAAAACCGACGATCCCAAATTCATTGATTATATAAATTTAATAATTAAAGCAACACACTCCGAAGAACTACCAGAAAATAGCACTGTTGTAAATTACAGAAAAACTATGCGCAGCGGTACTATACACCCCATTAAAAAAGACATATATATTTATGACAACAAAAAATTTACTCTATACGATAGATACATATATGGATACGATAATAACTATGTTAATTTTTATGAAGAGAAAAATGAAAAAGAGAAGGAATACGAAGAAGAAGACGACAAGGCGTCTAGTTTATGTGAAAATAAAATTATATTGTCGCAAATTAACTGTGAATCATTTGAAAATGATTTTAAATATTACCTCAGCGATTATAACTACGCGTTTTCAATTATAGATAATACTACAAATGTTCTTGTTGCGTTTGGTTTGTATCGTTAA Protein Sequence MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGVIELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPIKKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQINCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR
6xHis	—	6xHis affinity tag	1967..1985	Fwd
DNA Sequence CATCATCACCACCATCAC Protein Sequence HHHHHH
thrombin site	—	thrombin recognition and cleavage site	1994..2012	Fwd
DNA Sequence CTGGTGCCGCGCGGCAGC Protein Sequence LVPRGS
mTurquoise2	—	enhanced monomeric variant of CFP (Goed…	2036..2753	Fwd
DNA Sequence ATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCCTGTCCTGGGGCGTGCAGTGCTTCGCCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACTTTAGCGACAACGTCTATATCACCGCCGACAAGCAGAAGAACGGCATCAAGGCCAACTTCAAGATCCGCCACAACATCGAGGACGGCGGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCACCCAGTCCAAGCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTCTACAAG Protein Sequence MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK
P2A	—	2A peptide from porcine teschovirus-1 p…	2873..2930	Fwd
DNA Sequence GCTACTAACTTCAGCCTGCTGAAGCAGGCTGGAGACGTGGAGGAGAACCCTGGACCT Protein Sequence ATNFSLLKQAGDVEENPGP
mScarlet3	—	bright cysteine-free monomeric red fluo…	2954..3641	Fwd
DNA Sequence ATGGATAGCACCGAGGCAGTGATCAAGGAGTTCATGCGGTTCAAGGTGCACATGGAGGGCTCCATGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCTACGAGGGCACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCCCTGCCCTTCTCCTGGGACATCCTGTCCCCTCAGTTCATGTACGGCTCCAGGGCCTTCATCAAGCACCCCGCCGACATCCCCGACTACTGGAAGCAGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGATGATCTTCGAGGACGGCGGCACCGTGTCCGTGACCCAGGACACCTCCCTGGAGGACGGCACCCTGATCTACAAGGTGAAGCTCCGCGGCGGCAACTTCCCTCCTGACGGCCCCGTAATGCAGAAGAGGACAATGGGCTGGGAAGCATCCACCGAGCGGTTGTACCCCGAGGACGTCGTGCTGAAGGGCGACATTAAGATGGCCCTGCGCCTGAAGGACGGCGGCCGCTACCTGGCGGACTTCAAGACCACCTACAAGGCCAAGAAGCCCGTGCAGATGCCCGGCGCCTTCAACATCGACCGCAAGTTGGACATCACCTCCCACAACGAGGACTACACCGTGGTGGAACAGTACGAACGCTCCGTGGCCCGCCACTCCACCGGCGGCTCCGGTGGCTCC Protein Sequence MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYWKQSFPEGFKWERVMIFEDGGTVSVTQDTSLEDGTLIYKVKLRGGNFPPDGPVMQKRTMGWEASTERLYPEDVVLKGDIKMALRLKDGGRYLADFKTTYKAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHSTGGSGGS
ORF1629	—	baculovirus capsid-associated protein	4124..4713	Rev
DNA Sequence AATTCCAAAAAGCTCAACTACAAATTTGATCGCGGACGTGTTAGCCGACACAATTAATAGGCGTCGTGTGGCTATGGCAAAATCGTCTTCGGAAGCAACTTCTAACGACGAGGGTTGGGACGACGACGATAATCGGCCTAATAAAGCTAACACGCCCGATGTTAAATATGTCCAAGCTCTATTTAACGTGTTTACGTCGAGTCAATTGTATACTAACGACAGTGATGAAAGAAATACAAAAGCGCATAATATTTTGAACGACGTCGAACCTTTATTACAAAACAAAACACAAACGAATATCGACAAAGCTAGATTGCTGCTACAAGATTTGGCAAGTTTTGTGGCGTTGAGCGAAAATCCATTAGATAGTCCAGCCATCGGTTCGGAAAAACAACCCTTGTTTGAAACTAATCGAAACCTATTTTACAAATCTATTGAGGATTTAATATTTAAATTCAGATATAAAGACGCTGAAAATCATTTGATTTTCGCTCTAACATACCACCCTAAAGATTATAAATTTAATGAATTATTAAAATACGTACAACAATTGTCTGTAAATCAACAACGCACAGAATCTAGCGCTTAA Protein Sequence IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNRPNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKARLLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIFALTYHPKDYKFNELLKYVQQLSVNQQRTESSA
KanR	aph(3')-Ia	aminoglycoside phosphotransferase	5522..6338	Rev
DNA Sequence ATGAGCCATATTCAACGGGAAACGTCTTGCTCGAGGCCGCGATTAAATTCCAACATGGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGATTGTATGGGAAGCCCGATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGGCTAAACTGGCTGACGGAATTTATGCCTCTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCAGGGAAAACAGCATTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCATTCGATTCCTGTTTGTAATTGTCCTTTTAACGGCGATCGCGTATTTCGTCTCGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTGGTGCGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAGCTTTTGCCATTCTCACCGGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACGAGTCGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTTTTCAAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCACTTGATGCTCGATGAGTTTTTCTAA Protein Sequence MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF

enhancer 1

Label	Gene	Product	Position	Strand
CMV enhancer	—	—	919..1223	Fwd
DNA Sequence CGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTCCGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTACGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATG

polyA_signal 1

Label	Gene	Product	Position	Strand
beta-globin poly(A) signal	—	—	3804..3860	Fwd
DNA Sequence AATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCA

promoter 2

Label	Gene	Product	Position	Strand
CMV promoter	—	—	1224..1427	Fwd
DNA Sequence TGATGCGGTTTTGGCAGTACACCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAATAACCCCGCCCCGTTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGACGT
rhaB promoter	—	—	1774..1893	Fwd
DNA Sequence CACCACAATTCAGCAAATTGTGAACATCATCACGTTCATCTTTCCCTGGTTGCCAAAGGCCCATTTTCCTGTCAGTAACGAGAAGGTCGCGAATTCAGGCGCTTTTTAGACTGGTCGTA

protein_bind 1

Label	Gene	Product	Position	Strand
loxP	—	—	4721..4755	Rev
DNA Sequence ATAACTTCGTATAATGTATGCTATACGAAGTTAT

rep_origin 1

Label	Gene	Product	Position	Strand
ori	—	—	4922..5510	Rev
DNA Sequence TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGCGGAGCCTATGGAAA

terminator 1

Label	Gene	Product	Position	Strand
T7 terminator	—	—	3947..3995	Fwd
DNA Sequence CTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG