pDx_FusionRed-SYFP2

Plasmid Information

Purpose

Dual expression vector for bacteria and mammalian cells producing FusionRed-SYFP2 fusion protein for FRET analysis

Depositor

Dorus Gadella

Article

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

Insert

FusionRed-SYFP2

Sequence Information

Length

6,355 bp

Topology

circular

Molecule Type

other DNA

View on AddGene.org

Features

3'UTR 1

Label	Gene	Product	Position	Strand
polyhedrin 3' UTR	—	—	2281..2654	Fwd
DNA Sequence AACACGATACATTGTTATTAGTACATTTATTAAGCGCTAGATTCTGTGCGTTGTTGATTTACAGACAATTGTTGTACGTATTTTAATAATTCATTAAATTTATAATCTTTAGGGTGGTATGTTAGAGCGAAAATCAAATGATTTTCAGCGTCTTTATATCTGAATTTAAATATTAAATCCTCAATAGATTTGTAAAATAGGTTTCGATTAGTTTCAAACAAGGGTTGTTTTTCCGAACCGATGGCTGGACTATCTAATGGATTTTCGCTCAACGCCACAAAACTTGCCAAATCTTGTAGCAGCAATCTAGCTTTGTCGATATTCGTTTGTGTTTTGTTTTGTAATAAAGGTTCGACGTCGTTCAAAATATTAT

CDS 8

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
FusionRed	—	monomeric red fluorescent protein optim…	423..1119	Fwd
DNA Sequence ATGGTGAGCGAGCTGATTAAGGAGAACATGCCCATGAAGCTCTACATGGAGGGCACCGTGAACAACCACCACTTCAAGTGCACATCCGAGGGCGAAGGCAAGCCCTACGAGGGCACCCAGACCATGAGAATCAAGGTCGTCGAGGGCGGCCCTCTCCCCTTCGCCTTCGACATCCTGGCTACCAGCTTCATGTACGGCAGCAGAACCTTCATCAAGCACCCTCCGGGCATCCCCGACTTCTTTAAGCAGTCCTTCCCTGAGGGCTTCACATGGGAGAGAGTCACCACATACGAAGACGGGGGCGTGCTGACCGCTACCCAGGACACCAGCCTCCAGGACGGCTGCCTCATCTACAACGTCAAGGTTAGAGGGGTGAACTTCCCAGCCAACGGCCCTGTGATGCAGAAGAAAACACTCGGCTGGGAGGCCTCCACCGAGACGATGTACCCCGCTGACGGCGGCCTGGAAGGCGCATGTGACATGGCCCTGAAGCTCGTGGGCGGGGGCCACCTGATCTGCAACCTTGAGACCACATACAGATCCAAGAAACCCGCTACGAACCTCAAGATGCCCGGCGTCTACAACGTGGACCACAGACTGGAAAGAATCAAGGAGGCCGACGATGAGACCTACGTCGAGCAGCACGAGGTGGCTGTGGCCAGATACTCTACTGGTGGCGCTGGTGATCTGTACAAG Protein Sequence MVSELIKENMPMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKVVEGGPLPFAFDILATSFMYGSRTFIKHPPGIPDFFKQSFPEGFTWERVTTYEDGGVLTATQDTSLQDGCLIYNVKVRGVNFPANGPVMQKKTLGWEASTETMYPADGGLEGACDMALKLVGGGHLICNLETTYRSKKPATNLKMPGVYNVDHRLERIKEADDETYVEQHEVAVARYSTGGAGDLYK
SYFP2	—	monomeric optimized variant of EYFP, id…	1119..1836	Fwd
DNA Sequence ATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGCTGATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCCTGGGCTACGGCGTGCAGTGCTTCGCCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCTATATCACCGCCGACAAGCAGAAGAACGGCATCAAGGCCAACTTCAAGATCCGCCACAACATCGAGGACGGCGGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCTACCAGTCCAAGCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTCTACAAG Protein Sequence MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK
ORF1629	—	baculovirus capsid-associated protein	2310..2899	Rev
DNA Sequence AATTCCAAAAAGCTCAACTACAAATTTGATCGCGGACGTGTTAGCCGACACAATTAATAGGCGTCGTGTGGCTATGGCAAAATCGTCTTCGGAAGCAACTTCTAACGACGAGGGTTGGGACGACGACGATAATCGGCCTAATAAAGCTAACACGCCCGATGTTAAATATGTCCAAGCTCTATTTAACGTGTTTACGTCGAGTCAATTGTATACTAACGACAGTGATGAAAGAAATACAAAAGCGCATAATATTTTGAACGACGTCGAACCTTTATTACAAAACAAAACACAAACGAATATCGACAAAGCTAGATTGCTGCTACAAGATTTGGCAAGTTTTGTGGCGTTGAGCGAAAATCCATTAGATAGTCCAGCCATCGGTTCGGAAAAACAACCCTTGTTTGAAACTAATCGAAACCTATTTTACAAATCTATTGAGGATTTAATATTTAAATTCAGATATAAAGACGCTGAAAATCATTTGATTTTCGCTCTAACATACCACCCTAAAGATTATAAATTTAATGAATTATTAAAATACGTACAACAATTGTCTGTAAATCAACAACGCACAGAATCTAGCGCTTAA Protein Sequence IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNRPNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKARLLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIFALTYHPKDYKFNELLKYVQQLSVNQQRTESSA
KanR	aph(3')-Ia	aminoglycoside phosphotransferase	3708..4524	Rev
DNA Sequence ATGAGCCATATTCAACGGGAAACGTCTTGCTCGAGGCCGCGATTAAATTCCAACATGGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGATTGTATGGGAAGCCCGATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGGCTAAACTGGCTGACGGAATTTATGCCTCTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCAGGGAAAACAGCATTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCATTCGATTCCTGTTTGTAATTGTCCTTTTAACGGCGATCGCGTATTTCGTCTCGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTGGTGCGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAGCTTTTGCCATTCTCACCGGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACGAGTCGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTTTTCAAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCACTTGATGCTCGATGAGTTTTTCTAA Protein Sequence MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF
lef2	—	baculovirus late expression factor 2	4766..4891	Fwd
DNA Sequence TGCATCTTTTAATCAAATCCCAAGATGTGTATAAACCACCAAACTGCCAAAAAATGAAAACTGTCGACAAGCTCTGTCCGTTTGCTGGCAACTGCAAGGGTCTCAATCCTATTTGTAATTATTGA Protein Sequence HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY
ORF603	—	baculovirus ORF603 protein	4928..5534	Rev
DNA Sequence ATGGCTGTTATTTTTAACAACAAACAACTGCTCGCAGACAATAGTATAGAAAAGGGAGGTGAACTGTTTTTGTTTAACGGTTCGTACAACATTTTGGAAAGTTATGTTAATCCGGTGCTGCTAAAAAATGGTGTAATTGAACTAGAAGAAGCTGCGTACTATGCCGGCAACATATTGTATAAAACCGACGATCCCAAATTCATTGATTATATAAATTTAATAATTAAAGCAACACACTCCGAAGAACTACCAGAAAATAGCACTGTTGTAAATTACAGAAAAACTATGCGCAGCGGTACTATACACCCCATTAAAAAAGACATATATATTTATGACAACAAAAAATTTACTCTATACGATAGATACATATATGGATACGATAATAACTATGTTAATTTTTATGAAGAGAAAAATGAAAAAGAGAAGGAATACGAAGAAGAAGACGACAAGGCGTCTAGTTTATGTGAAAATAAAATTATATTGTCGCAAATTAACTGTGAATCATTTGAAAATGATTTTAAATATTACCTCAGCGATTATAACTACGCGTTTTCAATTATAGATAATACTACAAATGTTCTTGTTGCGTTTGGTTTGTATCGTTAA Protein Sequence MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGVIELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPIKKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQINCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR

enhancer 1

Label	Gene	Product	Position	Strand
CMV enhancer	—	—	5661..5965	Fwd
DNA Sequence CGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTCCGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTACGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATG

polyA_signal 1

Label	Gene	Product	Position	Strand
beta-globin poly(A) signal	—	—	1990..2046	Fwd
DNA Sequence AATAAAGGAAATTTATTTTCATTGCAATAGTGTGTTGGAATTTTTTGTGTCTCTCA

promoter 2

Label	Gene	Product	Position	Strand
rhaB promoter	—	—	161..280	Fwd
DNA Sequence CACCACAATTCAGCAAATTGTGAACATCATCACGTTCATCTTTCCCTGGTTGCCAAAGGCCCATTTTCCTGTCAGTAACGAGAAGGTCGCGAATTCAGGCGCTTTTTAGACTGGTCGTA
CMV promoter	—	—	5966..6169	Fwd
DNA Sequence TGATGCGGTTTTGGCAGTACACCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAATAACCCCGCCCCGTTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGACGT

protein_bind 1

Label	Gene	Product	Position	Strand
loxP	—	—	2907..2941	Rev
DNA Sequence ATAACTTCGTATAATGTATGCTATACGAAGTTAT

rep_origin 1

Label	Gene	Product	Position	Strand
ori	—	—	3108..3696	Rev
DNA Sequence TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGCGGAGCCTATGGAAA

terminator 1

Label	Gene	Product	Position	Strand
T7 terminator	—	—	2133..2181	Fwd
DNA Sequence CTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG