pCasSA

$193.00

  • Model: PVT11366
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pCasSA

PVT11366   2ug

 

pCasSA Information

Prokaryotic resistance: kanamycin Kan

Screening marker: chloramphenicol Chl

Clone strain: Escherichia coli Stbl3

Culture conditions: 30 C

 

pCasSA Description

PCasSA is a Staphylococcus aureus temperature sensitive gene editing plasmid.

 

pCasSA Sequence

 

LOCUS       Exported               10239 bp ds-DNA     circular SYN 07-JAN-2018
DEFINITION  synthetic circular DNA
FEATURES             Location/Qualifiers
     source          1..10239
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      78..666
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_RNA        804..879
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             1120..5226
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6793..7392
                     /codon_start=1
                     /label=repC
                     /translation="MSENVIKETENKKNSRGRNWTFVLYPESAKAEWLEYLKELHIQFV
                     VSPLHDRDTDTEDRMKKEHYHILVMYEGNKSYEQIKIITEELNATIPQIAGSVKGLVRY
                     MLHMDDPNKFKYQKEDMIVYGGVDVDELLKKTTTDRYKLIKEMIEFIDEQGIVEFKSLM
                     DYAMKFKFDDWFPLLCDNSAYVIQEYIKSNRYKSDR"
     CDS             8288..8938
                     /codon_start=1
                     /label=Chl
                     /translation="MKFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     CDS             9311..10105
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=Kan
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     terminator      10136..10230
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 gtcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
       61 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
      121 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
      181 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
      241 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
      301 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
      361 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
      421 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
      481 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
      541 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
      601 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
      661 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
      721 cacatgttct ttcctgcgtt gtcgacagag tttgcaaaat atacagggga ttatatataa
      781 tggaaacgag accattggtc tcagttttag agctagaaat agcaagttaa aataaggcta
      841 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttgaga tctgtccata
      901 cccatggtct agaatgctcg agtcagaaaa atatacctgt atcttttttc aaaagcaaac
      961 atgctttggg taaacatgta ggtattaacg tcaatgcgac aatagtagca ttgattaaat
     1021 gagaattagt aagtgtttta cttactaaat tttatttaac ctaaaaatga accacctgga
     1081 tgtgtgggat taaaaagtga agagaggagg acatatcaca tggataagaa atactcaata
     1141 ggcttagata tcggcacaaa tagcgtcgga tgggcggtga tcactgatga atataaggtt
     1201 ccgtctaaaa agttcaaggt tctgggaaat acagaccgcc acagtatcaa aaaaaatctt
     1261 ataggggctc ttttatttga cagtggagag acagcggaag cgactcgtct caaacggaca
     1321 gctcgtagaa ggtatacacg tcggaagaat cgtatttgtt atctacagga gattttttca
     1381 aatgagatgg cgaaagtaga tgatagtttc tttcatcgac ttgaagagtc ttttttggtg
     1441 gaagaagaca agaagcatga acgtcatcct atttttggaa atatagtaga tgaagttgct
     1501 tatcatgaga aatatccaac tatctatcat ctgcgaaaaa aattggtaga ttctactgat
     1561 aaagcggatt tgcgcttaat ctatttggcc ttagcgcata tgattaagtt tcgtggtcat
     1621 tttttgattg agggagattt aaatcctgat aatagtgatg tggacaaact atttatccag
     1681 ttggtacaaa cctacaatca attatttgaa gaaaacccta t
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