LentiCas9- EGFP

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  • Model: PVT10979
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LentiCas9-EGFP


Catalog No. PVT10979
Packing 2ug

 

LentiCas9-EGFP Information

Function Mammal Editing plasmids
Resistance Amp
Screen Zeo
Strain Stbl3
Culture temperature 37degrees centigrade

Promoter: EF-1 alpha core promoter

Copier: Ori, F1 ori, SV40 ori

Terminator: WPRE

Plasmid classification: mammalian cell plasmid; mammalian editing plasmid; mammalian Cas9 plasmid

Plasmid size: 13177 BP

Plasmid tags: C-nucleoplasmin NLS, C-FLAG, C-EGFP

Prokaryotic resistance: ampicillin Amp

Screening Marker: Bleomycin Zeo

Cloning strain: Escherichia coli Stbl3

Culture conditions: 37 C, aerobic, LB

Expressing host: mammalian cells such as 293T

 

LentiCas9-EGFP Description

Expresses human codon-optimized S. pyogenes Cas9 protein and EGFP from EFS promoter. 3rd generation lentiviral backbone.

 

LentiCas9-EGFP Reference

Genome-wide CRISPR screen in a mouse model of tumor growth and metastasis.

 

LentiCas9-EGFP Sequence

LOCUS       Exported               13177 bp ds-DNA    circular SYN 17-AUG-2017

KEYWORDS    lentiCas9-EGFP

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 13177)

FEATURES             Location/Qualifiers

     source          1..13177

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     enhancer        238..617

                     /note="CMV enhancer"

                     /note="human cytomegalovirus immediate early enhancer"

     promoter        619..817

                     /note="CMV promoter"

                     /note="human cytomegalovirus (CMV) immediate early 

                     promoter"

     LTR             835..1015

                     /note="5' LTR (truncated)"

                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"

     misc_feature    1062..1187

                     /note="HIV-1 Psi"

                     /note="packaging signal of human immunodeficiency virus 

                     type 1"

     misc_feature    1680..1913

                     /note="RRE"

                     /note="The Rev response element (RRE) of HIV-1 allows for 

                     Rev-dependent mRNA export from the nucleus to the 

                     cytoplasm."

     misc_feature    2440..2557

                     /note="cPPT/CTS"

                     /note="central polypurine tract and central termination 

                     sequence of HIV-1"

     promoter        2650..2861

                     /note="EF-1-alpha core promoter"

                     /note="core promoter for human elongation factor 

                     EF-1-alpha"

     CDS             2886..6989

                     /codon_start=1

                     /locus_tag="

                     "

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

                     pyogenes Type II CRISPR/Cas system"

                     /note="Cas9"

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

                     /protein_id="

                     "

                     /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             6990..7037

                     /codon_start=1

                     /product="bipartite nuclear localization signal from 

                     nucleoplasmin"

                     /note="nucleoplasmin NLS"

                     /translation="KRPAATKKAGQAKKKK"

     CDS             7038..7061

                     /codon_start=1

                     /product="FLAG(R) epitope tag, followed by an enterokinase 

                     cleavage site"

                     /note="FLAG"

                     /translation="DYKDDDDK"

     CDS             7071..7127

                     /codon_start=1

                     /product="2A peptide from porcine teschovirus-1 

                     polyprotein"

                     /note="P2A"

                     /note="Eukaryotic ribosomes fail to insert a peptide bond 

                     between the Gly and Pro residues, yielding separate 

                     polypeptides."

                     /translation="ATNFSLLKQAGDVEENPGP"

     CDS             7128..7844

                     /codon_start=1

                     /product="enhanced GFP"

                     /note="EGFP"

                     /note="mammalian codon-optimized"

                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK

                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG

                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA

                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE

                     FVTAAGITLGMDELYK"

     misc_feature    7869..8457

                     /note="WPRE"

                     /note="woodchuck hepatitis virus posttranscriptional 

                     regulatory element"

     LTR             8982..9162

                     /note="5' LTR (truncated)"

                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"

     polyA_signal    9194..9418

                     /note="bGH poly(A) signal"

                     /note="bovine growth hormone polyadenylation signal"

     rep_origin      9464..9892

                     /direction=RIGHT

                     /note="f1 ori"

                     /note="f1 bacteriophage origin of replication; arrow 

                     indicates direction of (+) strand synthesis"

     promoter        9906..10235

                     /note="SV40 promoter"

                     /note="SV40 enhancer and early promoter"

     rep_origin      10086..10221

                     /note="SV40 ori"

                     /note="SV40 origin of replication"

     promoter        10283..10330

                     /note="EM7 promoter"

                     /note="synthetic bacterial promoter "

     CDS             10349..10723

                     /codon_start=1

                     /gene="Sh ble from Streptoalloteichus hindustanus"

                     /product="antibiotic-binding protein"

                     /note="Zeo"

                     /note="confers resistance to bleomycin, phleomycin, and 

                     Zeocin(TM)"

                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD

                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR

                     EFALRDPAGNCVHFVAEEQD"

     polyA_signal    10853..10974

                     /note="SV40 poly(A) signal"

                     /note="SV40 polyadenylation signal"

     rep_origin      complement(11425..12013)

                     /direction=LEFT

                     /note="ori"

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

                     replication"

     CDS             complement(12184..13044)

                     /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"

     promoter        complement(13045..13149)

                     /gene="bla"

                     /note="AmpR promoter"

ORIGIN

        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg

       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt

      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc

      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac

      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat

      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg

      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt

      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag

      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc

      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag

      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt

      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc

      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg

      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct

      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt

      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac

      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc

     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc

     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa

     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg

     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata

     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc

     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga

     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc

     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca

     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg

     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga

     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata

     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg

     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg

     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag

     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt

     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt

     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt

     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag

     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata

     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta

     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta

     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa

     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt

     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat

     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa

     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag

     2581 agatccagtt tggttaatta gctagctagg tcttgaaagg agtgggaatt ggctccggtg

     2641 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg

     2701 gcaattgatc cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt

     2761 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg

     2821 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggaccggttc tagagcgctg

     2881 ccaccatgga caagaagtac agcatcggcc tggacatcgg caccaactct gtgggctggg

     2941 ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg

     3001 accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag

     3061 ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga

     3121 tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc

     3181 acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct

     3241 tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga

     3301 gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg

     3361 cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca

     3421 gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa

     3481 accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga

     3541 gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg

     3601 gaaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg

     3661 ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc

     3721 tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg

     3781 ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg

     3841 cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg

     3901 tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct

     3961 acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca

     4021 tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc

     4081 tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc

     4141 tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa

     4201 agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa

     4261 acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg

     4321 aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg

     4381 ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca

     4441 ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct

     4501 tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag

     4561 tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg

     4621 aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga

     4681 aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata

     4741 tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct

     4801 atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct

     4861 ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa

     4921 tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg

     4981 acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata

     5041 gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc

     5101 agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca

     5161 tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg

     5221 agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac

     5281 accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg

     5341 ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg

     5401 accatatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca

     5461 gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga

     5521 tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca

     5581 atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga

     5641 gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga

     5701 tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga

     5761 agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca

     5821 acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca

     5881 aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc

     5941 ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct

     6001 acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga

     6061 agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg

     6121 attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg

     6181 aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc

     6241 tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg

     6301 tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga

     6361 gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca

     6421 tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc

     6481 ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg

     6541 aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg

     6601 ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg

     6661 tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga

     6721 gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg

     6781 ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg

     6841 gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca

     6901 ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac

     6961 ggatcgacct gtctcagctg ggaggcgaca agcgacctgc cgccacaaag aaggctggac

     7021 aggctaagaa gaagaaagat tacaaagacg atgacgataa gggatccggc gcaacaaact

     7081 tctctctgct gaaacaagcc ggagatgtcg aagagaatcc tggaccggtg agcaagggcg

     7141 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc

     7201 acaagttcag cgtgtctggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga

     7261 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga

     7321 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca

     7381 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca

     7441 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc

     7501 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact

     7561 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggcgaact

     7621 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga

     7681 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt

     7741 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga

     7801 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagaattc gatatcaagc

     7861 ttatcggtaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact

     7921 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg

     7981 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg

     8041 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa

     8101 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc

     8161 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg

     8221 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt

     8281 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt

     8341 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc

     8401 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc

     8461 gataccgtcg acctcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag

     8521 ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag

     8581 tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact

     8641 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatatcc

     8701 ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaac tacacaccag

     8761 ggccagggat cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc

     8821 aagagaaggt agaagaagcc aatgaaggag agaacacccg cttgttacac cctgtgagcc

     8881 tgcatgggat ggatgacccg gagagagaag tattagagtg gaggtttgac agccgcctag

     8941 catttcatca catggcccga gagctgcatc cggactgtac tgggtctctc tggttagacc

     9001 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa

     9061 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga

     9121 gatccctcag acccttttag tcagtgtgga aaatctctag cagggcccgt ttaaacccgc

     9181 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg

     9241 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt

     9301 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc

     9361 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct

     9421 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc

     9481 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc

     9541 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc

     9601 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc

     9661 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg

     9721 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact

     9781 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt

     9841 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt

     9901 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc

     9961 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag

    10021 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc

    10081 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa

    10141 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt

    10201 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca

    10261 ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt

    10321 ataatacgac aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct

    10381 caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg

    10441 ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag

    10501 cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct

    10561 ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg

    10621 gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc

    10681 ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt

    10741 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg

    10801 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt

    10861 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc

    10921 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt

    10981 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt

    11041 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa

    11101 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc

    11161 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag

    11221 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt

    11281 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga

    11341 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg

    11401 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa

    11461 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt

    11521 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct

    11581 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct

    11641 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc

    11701 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt

    11761 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc

    11821 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat

    11881 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa

    11941 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa

    12001 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga

    12061 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct

    12121 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga

    12181 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc

    12241 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg

    12301 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat

    12361 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat

    12421 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg

    12481 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc

    12541 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa

    12601 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc

    12661 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt

    12721 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag

    12781 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt

    12841 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag

    12901 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac

    12961 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc

    13021 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca

    13081 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg

    13141 ggttccgcgc acatttcccc gaaaagtgcc acctgac

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