MiniCoopR U6:gRNA cdkn2a, mitfa:Cas9

118842 Danio rerio Bacterial Expression

Plasmid Information

Purpose
Targets cdkn2a and expresses zebrafish mitfa specifically in zebrafish melanocytes
Depositor
Leonard Zon
Article
Ablain et al Science. 2018 Nov 1. pii: science.aau650
Insert
cdkn2a gRNA (LOC100329528 Zebrafish)
Available Since
Nov. 28, 2018
Availability
Available

Sequence Information

Length
15,514 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 4
Label Gene Product Position Strand
AmpR bla beta-lactamase 124..985 Fwd
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
SV40 NLS — nuclear localization signal of SV40 (si… 8236..8257 Rev
DNA Sequence
CCTAAGAAGAAGAGAAAGGTG
Protein Sequence
PKKKRKV
Cas9 — Cas9 (Csn1) endonuclease from the Strep… 8272..12373 Rev
DNA Sequence
ATGGATAAGAAGTATAGCATCGGCCTGGATATTGGAACTAACTCCGTGGGTTGGGCAGTGATTACAGACGACTACAAGGTCCCTAGCAAGAAATTTAAGGTGCTGGGTAACACCGACAGGCACAGCATCAAGAAAAATCTGATTGGAGCCCTGCTGTTCGGTTCTGGAGAGACTGCCGAAGCAACACGCCTGAAAAGAACAGCAAGAAGGCGCTATACCAGAAGGAAGAATAGAATCTGTTACCTGCAGGAGATTTTCTCTAACGAAATGGCTAAGGTGGACGATTCATTCTTTCATAGGCTGGAGGAAAGTTTCCTGGTCGAGGAAGATAAGAAACACGAGCGCCATCCTATCTTTGGAAACATTGTGGACGAGGTCGCCTATCACGAAAAATACCCAACCATCTATCATCTGCGCAAGAAACTGGCTGACTCTACTGATAAAGCCGACCTGAGACTGATCTATCTGGCTCTGGCCCACATGATTAAGTTCAGGGGTCATTTTCTGATCGAGGGCGATCTGAACCCCGACAATTCCGATGTGGACAAGCTGTTCATCCAGCTGGTCCAGATTTACAATCAGCTGTTTGAGGAAAACCCTATTAATGCTTCCAGAGTGGACGCAAAAGCTATCCTGTCAGCCAGGCTGTCCAAGTCACGCAGACTGGAGAACCTGATTGCACAGCTGCCCGGAGAAAAGAGGAACGGTCTGTTTGGAAATCTGATCGCTCTGAGTCTGGGCCTGACTCCTAACTTCAAAAGCAATTTTGATCTGGCTGAGGACGCCAAACTGCAGCTGTCAAAGGACACATATGACGATGACCTGGATAACCTGCTGGCACAGATCGGAGATCAGTACGCTGACCTGTTCCTGGCTGCCAAAAATCTGTCCGACGCAATCCTGCTGTCAGATATTCTGAGAGTGAACAGCGAGATTACAAAAGCACCTCTGAGTGCCAGCATGATCAAGAGATATGACGAGCACCATCAGGATCTGACCCTGCTGAAGGCTCTGGTCAGGCAGCAGCTGCCAGAGAAGTACAAGGAAATTTTCTTTGATCAGTCCAAGAACGGCTACGCCGGTTATATCGACGGAGGCGCATCACAGGAGGAATTCTACAAGTTTATCAAACCTATTCTGGAGAAGATGGACGGAACTGAGGAACTGCTGGTGAAACTGAATAGAGAGGACCTGCTGAGGAAGCAGCGCACATTTGATAACGGTTCCATCCCACACCAGATTCATCTGGGAGAGCTGCACGCTATCCTGAGGCGCCAGGAAGACTTCTACCCCTTTCTGAAAGATAACCGCGAGAAGATCGAAAAAATTCTGACCTTCAGAATCCCTTACTATGTGGGTCCACTGGCTCGCGGAAACAGCAGATTTGCCTGGATGACTCGCAAATCCGAGGAAACCATTACTCCTTGGAACTTCGAGGAAGTGGTCGATAAGGGCGCCTCTGCACAGTCCTTCATCGAGAGAATGACTAATTTTGACAAAAACCTGCCCAATGAGAAAGTGCTGCCTAAGCACTCCCTGCTGTACGAGTATTTCACTGTCTATAACGAACTGACAAAGGTGAAATACGTCACCGAGGGCATGAGAAAGCCAGCCTTCCTGTCAGGAGAGCAGAAGAAAGCAATCGTGGATCTGCTGTTTAAAACCAACAGGAAAGTGACTGTCAAGCAGCTGAAGGAGGACTACTTCAAGAAAATTGAATGCTTCGATTCCGTGGAGATCAGCGGAGTCGAAGACAGATTTAACGCAAGCCTGGGCGCTTACCACGATCTGCTGAAGATCATTAAGGATAAAGACTTCCTGGACAACGAGGAAAATGAGGATATCCTGGAAGACATTGTGCTGACACTGACCCTGTTTGAGGACAGAGGAATGATCGAGGAAAGACTGAAAACCTATGCTCATCTGTTCGATGACAAGGTGATGAAACAGCTGAAGAGAAGGCGCTACACTGGCTGGGGTAGACTGAGCAGGAAGCTGATCAACGGCATTAGGGATAAACAGTCAGGAAAGACAATCCTGGACTTTCTGAAAAGTGATGGCTTCGCCAACCGCAATTTTATGCAGCTGATTCACGATGACAGTCTGACCTTCAAAGAGGACATCCAGAAGGCTCAGGTGTCTGGACAGGGCCACTCCCTGCATGAGCAGATTGCAAACCTGGCTGGAAGCCCAGCCATCAAGAAAGGCATTCTGCAGACAGTGAAAATCGTCGATGAGCTGGTGAAAGTCATGGGCCATAAGCCCGAAAACATCGTGATTGAGATGGCTCGCGAAAATCAGACAACCCAGAAGGGTCAGAAGAACAGTAGAGAGAGGATGAAAAGAATCGAGGAAGGCATTAAGGAGCTGGGTAGCCAGATCCTGAAAGAGCACCCAGTGGAAAACACACAGCTGCAGAATGAGAAGCTGTATCTGTACTATCTGCAGAATGGAAGAGATATGTACGTGGACCAGGAGCTGGATATTAACAGGCTGTCTGATTACGACGTGGATCATATCGTCCCCCAGAGTTTCATCAAAGATGACAGCATTGACAACAAGGTGCTGACCAGGTCCGACAAAAACAGAGGAAAATCAGATAATGTCCCTAGTGAGGAAGTGGTCAAGAAAATGAAGAACTACTGGAGACAGCTGCTGAATGCCAAACTGATCACTCAGAGGAAGTTTGATAACCTGACAAAAGCAGAGCGCGGTGGACTGTCAGAACTGGACAAAGCTGGATTCATCAAGAGGCAGCTGGTGGAAACACGCCAGATCACTAAACACGTCGCACAGATTCTGGATAGTCGCATGAACACAAAGTACGATGAGAATGACAAACTGATCAGAGAAGTGAAGGTCATTACCCTGAAGAGTAAACTGGTCAGCGACTTTAGGAAAGATTTCCAGTTTTATAAGGTCCGCGAGATTAACAATTATCACCATGCCCATGACGCATACCTGAACGCCGTGGTCGGTACCGCACTGATCAAGAAATACCCAAAACTGGAGAGCGAATTCGTGTACGGAGACTATAAGGTGTACGATGTCAGAAAAATGATCGCCAAGTCCGAGCAGGAAATTGGAAAAGCTACTGCCAAGTATTTCTTTTACTCAAACATCATGAATTTCTTTAAGACAGAGATCACCCTGGCCAATGGAGAAATCCGCAAAAGGCCCCTGATTGAGACAAACGGAGAGACAGGCGAAATCGTGTGGGACAAAGGCAGAGATTTTGCAACCGTGAGGAAGGTCCTGAGCATGCCTCAAGTGAATATCGTCAAGAAAACTGAGGTGCAGACAGGCGGTTTCTCAAAAGAAAGTATTCTGCCAAAACGCAACTCTGATAAGCTGATCGCTAGAAAGAAAGACTGGGACCCTAAGAAGTATGGAGGCTTTGACTCTCCCACTGTGGCATACTCCGTCCTGGTGGTCGCTAAGGTGGAGAAGGGCAAAAGCAAGAAACTGAAATCTGTCAAGGAGCTGCTGGGTATCACAATTATGGAGAGAAGCTCTTTCGAGAAGAACCCAATCGATTTTCTGGAGGCCAAAGGTTATAAGGAAGTGAAGAAAGACCTGATCATTAAACTGCCCAAGTACAGTCTGTTTGAGCTGGAAAACGGCAGGAAACGCATGCTGGCAAGCGCTGGAGAGCTGCAGAAAGGCAATGAACTGGCCCTGCCTTCTAAGTACGTGAACTTCCTGTATCTGGCAAGCCACTACGAGAAGCTGAAAGGATCTCCAGAGGATAACGAACAGAAACAGCTGTTTGTGGAGCAGCACAAGCATTATCTGGACGAGATCATTGAACAGATTAGCGAGTTCTCTAAAAGAGTGATCCTGGCCGACGCAAATCTGGATAAGGTCCTGTCTGCTTACAACAAACACAGAGATAAGCCCATCAGGGAGCAGGCCGAAAATATCATTCATCTGTTCACTCTGACAAACCTGGGCGCACCTGCAGCTTTCAAGTACTTCGACACTACAATCGATAGAAAGAGGTACACCTCCACTAAGGAGGTGCTGGACGCTACACTGATCCATCAGAGTATTACCGGCCTGTACGAAACAAGGATTGACCTGTCTCAGCTGGGTGGCGAC
Protein Sequence
MDKKYSIGLDIGTNSVGWAVITDDYKVPSKKFKVLGNTDRHSIKKNLIGALLFGSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLADSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQIYNQLFEENPINASRVDAKAILSARLSKSRRLENLIAQLPGEKRNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNSEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGAYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDRGMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGHSLHEQIANLAGSPAIKKGILQTVKIVDELVKVMGHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFIKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD
SV40 NLS — nuclear localization signal of SV40 (si… 12379..12400 Rev
DNA Sequence
CCTAAGAAGAAGAGAAAGGTG
Protein Sequence
PKKKRKV
RBS 1
Label Gene Product Position Strand
Shine-Dalgarno sequence — — 8351..8360 Fwd
DNA Sequence
ACCTCCTTA
polyA_signal 2
Label Gene Product Position Strand
SV40 poly(A) signal — — 7261..7396 Fwd
DNA Sequence
AACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGATC
SV40 poly(A) signal — — 7975..8110 Fwd
DNA Sequence
AACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGATC
promoter 5
Label Gene Product Position Strand
AmpR promoter bla — 19..124 Fwd
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
lac promoter — — 2067..2098 Fwd
DNA Sequence
TTTACACTTTATGCTTCCGGCTCGTATGTTG
T3 promoter — — 2166..2185 Fwd
DNA Sequence
AATTAACCCTCACTAAAGG
T3 promoter — — 8164..8183 Fwd
DNA Sequence
AATTAACCCTCACTAAAGG
T7 promoter — — 12453..12472 Rev
DNA Sequence
TAATACGACTCACTATAGG
protein_bind 6
Label Gene Product Position Strand
CAP binding site — — 2031..2053 Fwd
DNA Sequence
TAATGTGAGTTAGCTCACTCAT
lac operator — — 2105..2122 Fwd
DNA Sequence
TTGTGAGCGGATAACAA
attB3 mutant version of attB — 7892..7913 Fwd
DNA Sequence
CAACTATGTATAATAAAGTTG
attB2 mutant version of attB — 8111..8136 Fwd
DNA Sequence
CCCACTTTGTACAAGAAAGCTGGGT
attB1 mutant version of attB — 12499..12524 Rev
DNA Sequence
GCAAGTTTGTACAAAAAAGCAGGCT
attB4 mutant version of attB — 14397..14418 Rev
DNA Sequence
CAACTTTGTATAGAAAAGTTG
rep_origin 1
Label Gene Product Position Strand
ori — — 1155..1744 Fwd
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA