pX330A- 1x2

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  • Model: PVT10934
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pX330A-1x2


Catalog No. PVT10934
Packing 2ug

 

pX330A-1x2 Information

Replicon: PUC

Prokaryotic resistance: Amp

Clone strain: Stbl3

Culture conditions: 37 degrees

Plasmid host: Mammalian cells

Plasmid use: Gene editing

Fragment type: CRISPR

Fragment species:

Prokaryotic resistance: Amp

 

pX330A-1x2 Description
pX330A-1x2 is a Mammal Editing plasmids.

 

pX330A-1x2 Sequence

LOCUS       Exported                8964 bp ds-DNA     circular SYN 12-AUG-2018

DEFINITION  synthetic circular DNA

KEYWORDS    pX330A-1x2

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 8964)

  TITLE     Direct Submission

FEATURES             Location/Qualifiers

     source          1..8964

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     enhancer        8..293

                     /note="CMV enhancer"

                     /note="human cytomegalovirus immediate early enhancer; 

                     contains an 18-bp deletion relative to the standard CMV 

                     enhancer"

     promoter        295..572

                     /note="chicken beta-actin promoter"

     intron          573..801

                     /note="hybrid intron"

                     /note="hybrid between chicken beta-actin (CBA) and minute 

                     virus of mice (MMV) introns (Gray et al., 2011)"

     CDS             822..887

                     /codon_start=1

                     /product="three tandem FLAG(R) epitope tags, followed by an

                     enterokinase cleavage site"

                     /note="3xFLAG"

                     /translation="DYKDHDGDYKDHDIDYKDDDDK"

     CDS             894..914

                     /codon_start=1

                     /product="nuclear localization signal of SV40 large T 

                     antigen"

                     /note="SV40 NLS"

                     /translation="PKKKRKV"

     CDS             939..5039

                     /codon_start=1

                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 

                     pyogenes Type II CRISPR/Cas system"

                     /note="Cas9"

                     /note="generates RNA-guided double strand breaks in DNA"

                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN

                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF

                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF

                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE

                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI

                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ

                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR

                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN

                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF

                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS

                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL

                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ

                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH

                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL

                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS

                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV

                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN

                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE

                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE

                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT

                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL

                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA

                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD

                     ATLIHQSITGLYETRIDLSQLGGD"

     CDS             5040..5087

                     /codon_start=1

                     /product="bipartite nuclear localization signal from 

                     nucleoplasmin"

                     /note="nucleoplasmin NLS"

                     /translation="KRPAATKKAGQAKKKK"

     polyA_signal    5121..5328

                     /note="bGH poly(A) signal"

                     /note="bovine growth hormone polyadenylation signal"

     repeat_region   5337..5477

                     /note="AAV2 ITR"

                     /note="inverted terminal repeat of adeno-associated virus 

                     serotype 2"

     rep_origin      5552..6007

                     /direction=RIGHT

                     /note="f1 ori"

                     /note="f1 bacteriophage origin of replication; arrow 

                     indicates direction of (+) strand synthesis"

     promoter        6289..6393

                     /gene="bla"

                     /note="AmpR promoter"

     CDS             6394..7254

                     /codon_start=1

                     /gene="bla"

                     /product="beta-lactamase"

                     /note="AmpR"

                     /note="confers resistance to ampicillin, carbenicillin, and

                     related antibiotics"

                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI

                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS

                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW

                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA

                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS

                     LIKHW"

     rep_origin      7425..8013

                     /direction=RIGHT

                     /note="ori"

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 

                     replication"

     promoter        8075..8315

                     /note="U6 promoter"

                     /note="RNA polymerase III promoter for human U6 snRNA"

     misc_RNA        8342..8417

                     /note="gRNA scaffold"

                     /note="guide RNA scaffold for the Streptococcus pyogenes 

                     CRISPR/Cas9 system"

ORIGIN

        1 aggtacccgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc

       61 gcccattgac gtcaatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt

      121 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc

      181 ctattgacgt caatgacggt aaatggcccg cctggcattg tgcccagtac atgaccttat

      241 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg

      301 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt

      361 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggggcgcg

      421 cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg

      481 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc

      541 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgacgctg ccttcgcccc

      601 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc

      661 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagct gagcaagagg

      721 taagggttta agggatggtt ggttggtggg gtattaatgt ttaattacct ggagcacctg

      781 cctgaaatca ctttttttca ggttggaccg gtgccaccat ggactataag gaccacgacg

      841 gagactacaa ggatcatgat attgattaca aagacgatga cgataagatg gccccaaaga

      901 agaagcggaa ggtcggtatc cacggagtcc cagcagccga caagaagtac agcatcggcc

      961 tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca

     1021 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg

     1081 gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca

     1141 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg

     1201 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag

     1261 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc

     1321 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg

     1381 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc

     1441 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg

     1501 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca

     1561 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc

     1621 tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga

     1681 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg

     1741 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc

     1801 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga

     1861 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc

     1921 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag

     1981 agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc

     2041 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac

     2101 tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca

     2161 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt

     2221 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc

     2281 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga

     2341 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc

     2401 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc

     2461 ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat

     2521 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg

     2581 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact

     2641 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg

     2701 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca

     2761 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca

     2821 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga

     2881 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg

     2941 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg

     3001 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc

     3061 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg

     3121 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga

     3181 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga

     3241 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca

     3301 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg

     3361 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg

     3421 acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg

     3481 acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca

     3541 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg

     3601 ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga

     3661 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa

     3721 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc

     3781 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg

     3841 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc

     3901 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg

     3961 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa

     4021 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg

     4081 agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa

     4141 ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca

     4201 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt

     4261 ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta

     4321 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag

     4381 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca

     4441 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag

     4501 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg

     4561 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc

     4621 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc

     4681 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga

     4741 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca

     4801 aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata

     4861 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca

     4921 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc

     4981 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca

     5041 aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa gaaaaagtaa gaattcctag

     5101 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc

     5161 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga

     5221 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca

     5281 ggacagcaag ggggaggatt gggaagagaa tagcaggcat gctggggagc ggccgcagga

     5341 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg

     5401 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc

     5461 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat

     5521 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg

     5581 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct

     5641 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta

     5701 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa

     5761 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct

     5821 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc

     5881 aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg

     5941 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt

     6001 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc

     6061 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct

     6121 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca

     6181 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg

     6241 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct

     6301 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga

     6361 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc

     6421 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg

     6481 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc

     6541 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact

     6601 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc

     6661 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag

     6721 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat

     6781 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt

     6841 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa

     6901 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc

     6961 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg

     7021 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt

     7081 gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc actggggcca

     7141 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat

     7201 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca

     7261 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg

     7321 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg

     7381 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt

     7441 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg

     7501 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata

     7561 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca

     7621 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag

     7681 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc

     7741 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga

     7801 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg

     7861 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac

     7921 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg

     7981 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg

     8041 ttcctggcct tttgctggcc ttttgctcac atgtgagggc ctatttccca tgattccttc

     8101 atatttgcat atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa

     8161 cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc

     8221 agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt

     8281 cgatttcttg gctttatata tcttgtggaa aggacgaaac accgggtctt cgagaagacc

     8341 tgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag

     8401 tggcaccgag tcggtgcttt tttgttttag agctagaaat agcaagttaa aataaggcta

     8461 gtccgttttt agcgcgtgcg ccaattctgc agacaaatgg ctctagagac cggcgccgct

     8521 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg

     8581 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg

     8641 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta

     8701 atacgactca ctatagggcg aattgggtac cgggcccccc ctcgaggtcc tccagctttt

     8761 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg

     8821 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta

     8881 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg

     8941 ctttccaccg gtggtctctt ctag

//

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