pUFlip-floxed2A-KalTA4-; HE1A:BFP -2

180012 Danio rerio Bacterial Expression

Plasmid Information

Purpose
Donor for generating conditional alleles in zebrafish using precise CRISPR directed genomic integration with the GeneWeld method
Depositor
Jeffrey Essner
Article
Liu et al Elife. 2022 Jun 17;11. pii: 71478. doi:
Insert
2A-KalTA4; HE1A: BFP
Available Since
Feb. 7, 2022
Availability
Available

Sequence Information

Length
7,284 bp
Topology
circular
Molecule Type
other DNA

Features

CDS 6
Label Gene Product Position Strand
lacZ-alpha lacZ fragment LacZ-alpha fragment of beta-galactosida… 623..935 Fwd
DNA Sequence
ATGACCATGATTACGCCAAGCGCGCAATTAACCCTCACTAAAGGGAACAAAAGCTGGAGGACCTCGAGGGGGGGCCCGGTACCCAATTCGCCCTATAGTGAGTCGTATTACGCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCACCTGCCACACTTCCGGCCGCATAACTTTAAATAA
Protein Sequence
MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWHLPHFRPHNFK
P2A 2A peptide from porcine teschovirus-1 p… 1176..1233 Fwd
DNA Sequence
GCCACGAACTTCTCTCTGTTAAAGCAAGCAGGAGACGTGGAAGAAAACCCCGGTCCT
Protein Sequence
ATNFSLLKQAGDVEENPGP
GAL4 DNA binding domain Saccharomyces cerevisiae GAL4 (truncated) DNA binding domain of the GAL4 transcri… 1233..1674 Fwd
DNA Sequence
ATGAAACTGCTCTCATCCATCGAGCAAGCCTGCGACATTTGTCGGCTTAAGAAGCTGAAATGCTCCAAGGAAAAGCCGAAATGTGCCAAATGCCTGAAGAACAATTGGGAATGTCGTTACTCTCCCAAAACCAAGCGAAGTCCACTCACAAGGGCTCATCTGACCGAAGTGGAGAGCAGGCTAGAGAGACTGGAACAACTCTTTTTGCTCATCTTCCCTAGAGAGGACCTTGACATGATCCTCAAGATGGATTCTCTCCAGGATATTAAAGCCCTTTTGACTGGCTTATTCGTCCAGGACAATGTGAACAAAGACGCTGTGACAGACCGATTGGCAAGTGTCGAGACCGATATGCCTCTGACACTGAGACAGCACAGAATCAGCGCTACTTCCTCAAGCGAAGAGTCTTCTAACAAGGGACAGAGACAGCTGACTGTTTCG
Protein Sequence
MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKRSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKDAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS
SV40 NLS nuclear localization signal of SV40 (si… 3414..3435 Fwd
DNA Sequence
CCAAAGAAGAAGCGTAAGGTA
Protein Sequence
PKKKRKV
TagBFP monomeric blue fluorescent protein 3438..4140 Fwd
DNA Sequence
ATGAGCGAGCTGATTAAGGAGAACATGCACATGAAGCTGTACATGGAGGGCACCGTGGACAACCATCACTTCAAGTGCACATCCGAGGGCGAAGGCAAGCCCTACGAGGGCACCCAGACCATGAGAATCAAGGTGGTCGAGGGCGGCCCTCTCCCCTTCGCCTTCGACATCCTGGCTACTAGCTTCCTCTACGGCAGCAAGACCTTCATCAACCACACCCAGGGCATCCCCGACTTCTTCAAGCAGTCCTTCCCTGAGGGCTTCACATGGGAGAGAGTCACCACATACGAAGACGGGGGCGTGCTGACCGCTACCCAGGACACCAGCCTCCAGGACGGCTGCCTCATCTACAACGTCAAGATCAGAGGGGTGAACTTCACATCCAACGGCCCTGTGATGCAGAAGAAAACACTCGGCTGGGAGGCCTTCACCGAGACGCTGTACCCCGCTGACGGCGGCCTGGAAGGCAGAAACGACATGGCCCTGAAGCTCGTGGGCGGGAGCCATCTGATCGCAAACATCAAGACCACATATAGATCCAAGAAACCCGCTAAGAACCTCAAGATGCCTGGCGTCTACTATGTGGACTACAGACTGGAAAGAATCAAGGAGGCCAACAACGAGACCTACGTCGAGCAGCACGAGGTGGCAGTGGCCAGATACTGCGACCTCCCTAGCAAACTGGGGCACAAGCTCAATTAA
Protein Sequence
MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKLVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCDLPSKLGHKLN
KanR aph(3')-Ia aminoglycoside phosphotransferase 5795..6605 Fwd
DNA Sequence
ATGAGCCATATTCAACGGGAAACGTCGAGGCCGCGATTAAATTCCAACATGGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGCTTGTATGGGAAGCCCGATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGACTAAACTGGCTGACGGAATTTATGCCTCTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCCGGAAAAACAGCATTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCATTCGATTCCTGTTTGTAATTGTCCTTTTAACAGCGATCGCGTATTTCGTCTCGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTGATGCGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAACTTTTGCCATTCTCACCGGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACGAGTCGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTTTTCAAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCATTTGATGCTCGATGAGTTTTTCTAA
Protein Sequence
MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF
promoter 4
Label Gene Product Position Strand
lac promoter 549..580 Fwd
DNA Sequence
TTTACACTTTATGCTTCCGGCTCGTATGTTG
T3 promoter 648..667 Fwd
DNA Sequence
AATTAACCCTCACTAAAGG
T7 promoter 714..733 Rev
DNA Sequence
TAATACGACTCACTATAGG
T7 promoter 5490..5509 Fwd
DNA Sequence
TAATACGACTCACTATAGG
protein_bind 8
Label Gene Product Position Strand
CAP binding site 513..535 Fwd
DNA Sequence
TAATGTGAGTTAGCTCACTCAT
lac operator 587..604 Fwd
DNA Sequence
TTGTGAGCGGATAACAA
rox 922..954 Fwd
DNA Sequence
TAACTTTAAATAATGCCAATTATTTAAAGTTA
loxP 959..993 Rev
DNA Sequence
ATAACTTCGTATAATGTATGCTATACGAAGTTAT
lox2272 997..1031 Rev
DNA Sequence
ATAACTTCGTATAAAGTATCCTATACGAAGTTAT
loxP 5214..5248 Fwd
DNA Sequence
ATAACTTCGTATAATGTATGCTATACGAAGTTAT
lox2272 5253..5287 Fwd
DNA Sequence
ATAACTTCGTATAAAGTATCCTATACGAAGTTAT
rox 5293..5325 Fwd
DNA Sequence
TAACTTTAAATAATTGGCATTATTTAAAGTTA
rep_origin 1
Label Gene Product Position Strand
ori 0..7284 Fwd
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGGTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA