p104Tol2-tp1:Cas9-t2A-GFP, 4xU6:sgRNA

227773 Danio rerio Bacterial Expression

Plasmid Information

Purpose
Expression of tp1 (Notch) dependent Cas9-GFP and U6-driven 4 sgRNAs in zebrafish
Depositor
Bushra Raj
Article
Siniscalco et al Development. 2024 Nov 22:dev.203102. doi
Promoter
U6 and tp1
Available Since
Dec. 10, 2024
Availability
Available

Sequence Information

Length
12,061 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 7
Label Gene Product Position Strand
Cas9 — Cas9 (Csn1) endonuclease from the Strep… 0..12061 Fwd
DNA Sequence
ATGGATAAGAAGTATAGCATCGGCCTGGATATTGGAACTAACTCCGTGGGTTGGGCAGTGATTACAGACGACTACAAGGTCCCTAGCAAGAAATTTAAGGTGCTGGGTAACACCGACAGGCACAGCATCAAGAAAAATCTGATTGGAGCCCTGCTGTTCGGTTCTGGAGAGACTGCCGAAGCAACACGCCTGAAAAGAACAGCAAGAAGGCGCTATACCAGAAGGAAGAATAGAATCTGTTACCTGCAGGAGATTTTCTCTAACGAAATGGCTAAGGTGGACGATTCATTCTTTCATAGGCTGGAGGAAAGTTTCCTGGTCGAGGAAGATAAGAAACACGAGCGCCATCCTATCTTTGGAAACATTGTGGACGAGGTCGCCTATCACGAAAAATACCCAACCATCTATCATCTGCGCAAGAAACTGGCTGACTCTACTGATAAAGCCGACCTGAGACTGATCTATCTGGCTCTGGCCCACATGATTAAGTTCAGGGGTCATTTTCTGATCGAGGGCGATCTGAACCCCGACAATTCCGATGTGGACAAGCTGTTCATCCAGCTGGTCCAGATTTACAATCAGCTGTTTGAGGAAAACCCTATTAATGCTTCCAGAGTGGACGCAAAAGCTATCCTGTCAGCCAGGCTGTCCAAGTCACGCAGACTGGAGAACCTGATTGCACAGCTGCCCGGAGAAAAGAGGAACGGTCTGTTTGGAAATCTGATCGCTCTGAGTCTGGGCCTGACTCCTAACTTCAAAAGCAATTTTGATCTGGCTGAGGACGCCAAACTGCAGCTGTCAAAGGACACATATGACGATGACCTGGATAACCTGCTGGCACAGATCGGAGATCAGTACGCTGACCTGTTCCTGGCTGCCAAAAATCTGTCCGACGCAATCCTGCTGTCAGATATTCTGAGAGTGAACAGCGAGATTACAAAAGCACCTCTGAGTGCCAGCATGATCAAGAGATATGACGAGCACCATCAGGATCTGACCCTGCTGAAGGCTCTGGTCAGGCAGCAGCTGCCAGAGAAGTACAAGGAAATTTTCTTTGATCAGTCCAAGAACGGCTACGCCGGTTATATCGACGGAGGCGCATCACAGGAGGAATTCTACAAGTTTATCAAACCTATTCTGGAGAAGATGGACGGAACTGAGGAACTGCTGGTGAAACTGAATAGAGAGGACCTGCTGAGGAAGCAGCGCACATTTGATAACGGTTCCATCCCACACCAGATTCATCTGGGAGAGCTGCACGCTATCCTGAGGCGCCAGGAAGACTTCTACCCCTTTCTGAAAGATAACCGCGAGAAGATCGAAAAAATTCTGACCTTCAGAATCCCTTACTATGTGGGTCCACTGGCTCGCGGAAACAGCAGATTTGCCTGGATGACTCGCAAATCCGAGGAAACCATTACTCCTTGGAACTTCGAGGAAGTGGTCGATAAGGGCGCCTCTGCACAGTCCTTCATCGAGAGAATGACTAATTTTGACAAAAACCTGCCCAATGAGAAAGTGCTGCCTAAGCACTCCCTGCTGTACGAGTATTTCACTGTCTATAACGAACTGACAAAGGTGAAATACGTCACCGAGGGCATGAGAAAGCCAGCCTTCCTGTCAGGAGAGCAGAAGAAAGCAATCGTGGATCTGCTGTTTAAAACCAACAGGAAAGTGACTGTCAAGCAGCTGAAGGAGGACTACTTCAAGAAAATTGAATGCTTCGATTCCGTGGAGATCAGCGGAGTCGAAGACAGATTTAACGCAAGCCTGGGCGCTTACCACGATCTGCTGAAGATCATTAAGGATAAAGACTTCCTGGACAACGAGGAAAATGAGGATATCCTGGAAGACATTGTGCTGACACTGACCCTGTTTGAGGACAGAGGAATGATCGAGGAAAGACTGAAAACCTATGCTCATCTGTTCGATGACAAGGTGATGAAACAGCTGAAGAGAAGGCGCTACACTGGCTGGGGTAGACTGAGCAGGAAGCTGATCAACGGCATTAGGGATAAACAGTCAGGAAAGACAATCCTGGACTTTCTGAAAAGTGATGGCTTCGCCAACCGCAATTTTATGCAGCTGATTCACGATGACAGTCTGACCTTCAAAGAGGACATCCAGAAGGCTCAGGTGTCTGGACAGGGCCACTCCCTGCATGAGCAGATTGCAAACCTGGCTGGAAGCCCAGCCATCAAGAAAGGCATTCTGCAGACAGTGAAAATCGTCGATGAGCTGGTGAAAGTCATGGGCCATAAGCCCGAAAACATCGTGATTGAGATGGCTCGCGAAAATCAGACAACCCAGAAGGGTCAGAAGAACAGTAGAGAGAGGATGAAAAGAATCGAGGAAGGCATTAAGGAGCTGGGTAGCCAGATCCTGAAAGAGCACCCAGTGGAAAACACACAGCTGCAGAATGAGAAGCTGTATCTGTACTATCTGCAGAATGGAAGAGATATGTACGTGGACCAGGAGCTGGATATTAACAGGCTGTCTGATTACGACGTGGATCATATCGTCCCCCAGAGTTTCATCAAAGATGACAGCATTGACAACAAGGTGCTGACCAGGTCCGACAAAAACAGAGGAAAATCAGATAATGTCCCTAGTGAGGAAGTGGTCAAGAAAATGAAGAACTACTGGAGACAGCTGCTGAATGCCAAACTGATCACTCAGAGGAAGTTTGATAACCTGACAAAAGCAGAGCGCGGTGGACTGTCAGAACTGGACAAAGCTGGATTCATCAAGAGGCAGCTGGTGGAAACACGCCAGATCACTAAACACGTCGCACAGATTCTGGATAGTCGCATGAACACAAAGTACGATGAGAATGACAAACTGATCAGAGAAGTGAAGGTCATTACCCTGAAGAGTAAACTGGTCAGCGACTTTAGGAAAGATTTCCAGTTTTATAAGGTCCGCGAGATTAACAATTATCACCATGCCCATGACGCATACCTGAACGCCGTGGTCGGTACCGCACTGATCAAGAAATACCCAAAACTGGAGAGCGAATTCGTGTACGGAGACTATAAGGTGTACGATGTCAGAAAAATGATCGCCAAGTCCGAGCAGGAAATTGGAAAAGCTACTGCCAAGTATTTCTTTTACTCAAACATCATGAATTTCTTTAAGACAGAGATCACCCTGGCCAATGGAGAAATCCGCAAAAGGCCCCTGATTGAGACAAACGGAGAGACAGGCGAAATCGTGTGGGACAAAGGCAGAGATTTTGCAACCGTGAGGAAGGTCCTGAGCATGCCTCAAGTGAATATCGTCAAGAAAACTGAGGTGCAGACAGGCGGTTTCTCAAAAGAAAGTATTCTGCCAAAACGCAACTCTGATAAGCTGATCGCTAGAAAGAAAGACTGGGACCCTAAGAAGTATGGAGGCTTTGACTCTCCCACTGTGGCATACTCCGTCCTGGTGGTCGCTAAGGTGGAGAAGGGCAAAAGCAAGAAACTGAAATCTGTCAAGGAGCTGCTGGGTATCACAATTATGGAGAGAAGCTCTTTCGAGAAGAACCCAATCGATTTTCTGGAGGCCAAAGGTTATAAGGAAGTGAAGAAAGACCTGATCATTAAACTGCCCAAGTACAGTCTGTTTGAGCTGGAAAACGGCAGGAAACGCATGCTGGCAAGCGCTGGAGAGCTGCAGAAAGGCAATGAACTGGCCCTGCCTTCTAAGTACGTGAACTTCCTGTATCTGGCAAGCCACTACGAGAAGCTGAAAGGATCTCCAGAGGATAACGAACAGAAACAGCTGTTTGTGGAGCAGCACAAGCATTATCTGGACGAGATCATTGAACAGATTAGCGAGTTCTCTAAAAGAGTGATCCTGGCCGACGCAAATCTGGATAAGGTCCTGTCTGCTTACAACAAACACAGAGATAAGCCCATCAGGGAGCAGGCCGAAAATATCATTCATCTGTTCACTCTGACAAACCTGGGCGCACCTGCAGCTTTCAAGTACTTCGACACTACAATCGATAGAAAGAGGTACACCTCCACTAAGGAGGTGCTGGACGCTACACTGATCCATCAGAGTATTACCGGCCTGTACGAAACAAGGATTGACCTGTCTCAGCTGGGTGGCGAC
Protein Sequence
MDKKYSIGLDIGTNSVGWAVITDDYKVPSKKFKVLGNTDRHSIKKNLIGALLFGSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLADSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQIYNQLFEENPINASRVDAKAILSARLSKSRRLENLIAQLPGEKRNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNSEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGAYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDRGMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGHSLHEQIANLAGSPAIKKGILQTVKIVDELVKVMGHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFIKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD
SV40 NLS — nuclear localization signal of SV40 (si… 2496..2517 Fwd
DNA Sequence
CCTAAGAAGAAGAGAAAGGTG
Protein Sequence
PKKKRKV
T2A — 2A peptide from Thosea asigna virus cap… 2532..2586 Fwd
DNA Sequence
GAGGGCAGAGGCAGTCTGCTGACATGCGGTGACGTGGAAGAGAATCCCGGCCCT
Protein Sequence
EGRGSLLTCGDVEENPGP
EGFP — the original enhanced GFP (Yang et al.,… 2592..3312 Fwd
DNA Sequence
ATGGTGAGCAAGGGCGAAGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCCTGACCTACGGCGTGCAGTGCTTCAGCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCTATATCATGGCCGACAAGCAGAAGAACGGCATCAAGGTGAACTTCAAGATCCGCCACAACATCGAGGACGGCAGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCACCCAGTCCGCCCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTGTACAAGTAA
Protein Sequence
MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK
EGFP — the original enhanced GFP (Yang et al.,… 2592..3309 Fwd
DNA Sequence
ATGGTGAGCAAGGGCGAAGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCCTGACCTACGGCGTGCAGTGCTTCAGCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCTATATCATGGCCGACAAGCAGAAGAACGGCATCAAGGTGAACTTCAAGATCCGCCACAACATCGAGGACGGCAGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCACCCAGTCCGCCCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTGTACAAG
Protein Sequence
MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK
AmpR bla beta-lactamase 7894..8755 Rev
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
SV40 NLS — nuclear localization signal of SV40 (si… 10414..10435 Fwd
DNA Sequence
CCTAAGAAGAAGAGAAAGGTG
Protein Sequence
PKKKRKV
RBS 1
Label Gene Product Position Strand
Shine-Dalgarno sequence — — 2393..2402 Fwd
DNA Sequence
TAAGGAGGT
polyA_signal 2
Label Gene Product Position Strand
SV40 poly(A) signal — — 3421..3556 Fwd
DNA Sequence
GATCCAGACATGATAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTT
SV40 poly(A) signal — — 5835..5970 Fwd
DNA Sequence
GATCCAGACATGATAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTT
promoter 5
Label Gene Product Position Strand
T3 promoter — — 3348..3367 Rev
DNA Sequence
AATTAACCCTCACTAAAGG
T3 promoter — — 6694..6713 Rev
DNA Sequence
AATTAACCCTCACTAAAGG
lac promoter — — 6781..6812 Rev
DNA Sequence
TTTACACTTTATGCTTCCGGCTCGTATGTTG
AmpR promoter bla — 8755..8860 Rev
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
T7 promoter — — 10342..10361 Fwd
DNA Sequence
TAATACGACTCACTATAGG
protein_bind 6
Label Gene Product Position Strand
attB2 mutant version of attB — 3395..3420 Rev
DNA Sequence
CCCACTTTGTACAAGAAAGCTGGGT
attB3 mutant version of attB — 3618..3639 Rev
DNA Sequence
CAACTATGTATAATAAAGTTG
lac operator — — 6757..6774 Fwd
DNA Sequence
TTGTTATCCGCTCACAA
CAP binding site — — 6826..6848 Fwd
DNA Sequence
ATGAGTGAGCTAACTCACATTA
attB4 mutant version of attB — 9975..9996 Fwd
DNA Sequence
CAACTTTGTATAGAAAAGTTG
attB1 mutant version of attB — 10290..10315 Fwd
DNA Sequence
ACAAGTTTGTACAAAAAAGCAGGCT
rep_origin 1
Label Gene Product Position Strand
ori — — 7135..7724 Rev
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA