pUFlip-floxed2A-KalTA4-; gcry1:BFP -2

173891 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; gcry1: BFP
Available Since
Aug. 20, 2021
Availability
Available

Sequence Information

Length
7,559 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… 669..981 Fwd
DNA Sequence
ATGACCATGATTACGCCAAGCGCGCAATTAACCCTCACTAAAGGGAACAAAAGCTGGAGGACCTCGAGGGGGGGCCCGGTACCCAATTCGCCCTATAGTGAGTCGTATTACGCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCACCTGCCACACTTCCGGCCGCATAACTTTAAATAA
Protein Sequence
MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWHLPHFRPHNFK
P2A 2A peptide from porcine teschovirus-1 p… 1222..1279 Fwd
DNA Sequence
GCCACGAACTTCTCTCTGTTAAAGCAAGCAGGAGACGTGGAAGAAAACCCCGGTCCT
Protein Sequence
ATNFSLLKQAGDVEENPGP
GAL4 DNA binding domain Saccharomyces cerevisiae GAL4 (truncated) DNA binding domain of the GAL4 transcri… 1279..1720 Fwd
DNA Sequence
ATGAAACTGCTCTCATCCATCGAGCAAGCCTGCGACATTTGTCGGCTTAAGAAGCTGAAATGCTCCAAGGAAAAGCCGAAATGTGCCAAATGCCTGAAGAACAATTGGGAATGTCGTTACTCTCCCAAAACCAAGCGAAGTCCACTCACAAGGGCTCATCTGACCGAAGTGGAGAGCAGGCTAGAGAGACTGGAACAACTCTTTTTGCTCATCTTCCCTAGAGAGGACCTTGACATGATCCTCAAGATGGATTCTCTCCAGGATATTAAAGCCCTTTTGACTGGCTTATTCGTCCAGGACAATGTGAACAAAGACGCTGTGACAGACCGATTGGCAAGTGTCGAGACCGATATGCCTCTGACACTGAGACAGCACAGAATCAGCGCTACTTCCTCAAGCGAAGAGTCTTCTAACAAGGGACAGAGACAGCTGACTGTTTCG
Protein Sequence
MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKRSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKDAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS
SV40 NLS nuclear localization signal of SV40 (si… 3735..3756 Fwd
DNA Sequence
CCAAAGAAGAAGCGTAAGGTA
Protein Sequence
PKKKRKV
TagBFP monomeric blue fluorescent protein 3759..4461 Fwd
DNA Sequence
ATGAGCGAGCTGATTAAGGAGAACATGCACATGAAGCTGTACATGGAGGGCACCGTGGACAACCATCACTTCAAGTGCACATCCGAGGGCGAAGGCAAGCCCTACGAGGGCACCCAGACCATGAGAATCAAGGTGGTCGAGGGCGGCCCTCTCCCCTTCGCCTTCGACATCCTGGCTACTAGCTTCCTCTACGGCAGCAAGACCTTCATCAACCACACCCAGGGCATCCCCGACTTCTTCAAGCAGTCCTTCCCTGAGGGCTTCACATGGGAGAGAGTCACCACATACGAAGACGGGGGCGTGCTGACCGCTACCCAGGACACCAGCCTCCAGGACGGCTGCCTCATCTACAACGTCAAGATCAGAGGGGTGAACTTCACATCCAACGGCCCTGTGATGCAGAAGAAAACACTCGGCTGGGAGGCCTTCACCGAGACGCTGTACCCCGCTGACGGCGGCCTGGAAGGCAGAAACGACATGGCCCTGAAGCTCGTGGGCGGGAGCCATCTGATCGCAAACATCAAGACCACATATAGATCCAAGAAACCCGCTAAGAACCTCAAGATGCCTGGCGTCTACTATGTGGACTACAGACTGGAAAGAATCAAGGAGGCCAACAACGAGACCTACGTCGAGCAGCACGAGGTGGCAGTGGCCAGATACTGCGACCTCCCTAGCAAACTGGGGCACAAGCTCAATTAA
Protein Sequence
MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKLVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCDLPSKLGHKLN
KanR aph(3')-Ia aminoglycoside phosphotransferase 6116..6926 Fwd
DNA Sequence
ATGAGCCATATTCAACGGGAAACGTCGAGGCCGCGATTAAATTCCAACATGGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGCTTGTATGGGAAGCCCGATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGACTAAACTGGCTGACGGAATTTATGCCTCTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCCGGAAAAACAGCATTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCATTCGATTCCTGTTTGTAATTGTCCTTTTAACAGCGATCGCGTATTTCGTCTCGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTGATGCGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAACTTTTGCCATTCTCACCGGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACGAGTCGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTTTTCAAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCATTTGATGCTCGATGAGTTTTTCTAA
Protein Sequence
MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF
promoter 4
Label Gene Product Position Strand
lac promoter 595..626 Fwd
DNA Sequence
TTTACACTTTATGCTTCCGGCTCGTATGTTG
T3 promoter 694..713 Fwd
DNA Sequence
AATTAACCCTCACTAAAGG
T7 promoter 760..779 Rev
DNA Sequence
TAATACGACTCACTATAGG
T7 promoter 5811..5830 Fwd
DNA Sequence
TAATACGACTCACTATAGG
protein_bind 8
Label Gene Product Position Strand
CAP binding site 559..581 Fwd
DNA Sequence
TAATGTGAGTTAGCTCACTCAT
lac operator 633..650 Fwd
DNA Sequence
TTGTGAGCGGATAACAA
rox 968..1000 Fwd
DNA Sequence
TAACTTTAAATAATGCCAATTATTTAAAGTTA
loxP 1005..1039 Rev
DNA Sequence
ATAACTTCGTATAATGTATGCTATACGAAGTTAT
lox2272 1043..1077 Rev
DNA Sequence
ATAACTTCGTATAAAGTATCCTATACGAAGTTAT
loxP 5535..5569 Fwd
DNA Sequence
ATAACTTCGTATAATGTATGCTATACGAAGTTAT
lox2272 5574..5608 Fwd
DNA Sequence
ATAACTTCGTATAAAGTATCCTATACGAAGTTAT
rox 5614..5646 Fwd
DNA Sequence
TAACTTTAAATAATTGGCATTATTTAAAGTTA
rep_origin 1
Label Gene Product Position Strand
ori 0..7559 Fwd
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGGTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA