LOCUS       sequence-178380-       12494 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for Cas9 knockout, lentiviral expression of Cas9. Alternate plasmid 
            name: pXR111.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12494)
  AUTHORS   Doench JG, Hartenian E, Graham DB, Tothova Z, Hegde M, Smith I, 
            Sullender M, Ebert BL, Xavier RJ, Root DE
  TITLE     Rational design of highly active sgRNAs for CRISPR-Cas9-mediated 
            gene inactivation.
  JOURNAL   Nat Biotechnol. 2014 Sep 3. doi: 10.1038/nbt.3026.
  PUBMED    25184501
REFERENCE   2  (bases 1 to 12494)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2014 Sep 3. doi: 10.1038/nbt.3026."
COMMENT     Sequence Label: pLX_311-Cas9
FEATURES             Location/Qualifiers
     source          1..12494
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             79..474
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    548..665
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        803..1981
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1034..1972
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1929..1949
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    2001..2025
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2028..2093
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2100..2120
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2145..6245
                     /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="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             2145..6245
                     /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="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             6246..6293
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    complement(6296..6320)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6328..6369
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    6420..7008
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6473..6493)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6891..6902)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7080..7313
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7391..7512
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7428..7447)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7482..7501
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7552..7687
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7707..7726)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7708..7726)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7736..7753)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7736..7752)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7745..7767)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7894..8349
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7981..8000)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8191..8212
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8375..8479
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8480..8548,8549..9340)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8698..8717)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9511..10099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10000..10019
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10253..10270
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10387..10408
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10423..10428,10429..10446,10447..10453)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10461..10477
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     10466..10488
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10485..10501
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10485..10501
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10520..10540
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10522..10540
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        10568..10794
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             10795..10975
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11022..11147
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11640..11873
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12058..12102
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        join(12333..12494,1..49)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        join(12355..12494,1..57)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      join(12402..12494,1..43)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
       61 ctttctagag gtaccaccat ggccaagcct ttgtctcaag aagaatccac cctcattgaa
      121 agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca
      181 gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga
      241 ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact
      301 tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga
      361 caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag
      421 ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga gggcctgcag
      481 ctgcagtagt aaggcgcgcc gttaacgaat tctagatctt gagacaaatg gcagtattca
      541 tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag
      601 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
      661 attttcgggt ttattacagg gacagcagag atccactttg gcgccggctc gaggcctgca
      721 ggtgcaaaga tggataaagt tttaaacaga gaggaatctt tgcagctaat ggaccttcta
      781 ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc
      841 cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg
      901 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
      961 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     1021 cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta
     1081 tggcccttgc gtgccttgaa ttacttccac ctggctgcag tacgtgattc ttgatcccga
     1141 gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc
     1201 tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca
     1261 ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc
     1321 tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac
     1381 tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg
     1441 ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc
     1501 tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc
     1561 aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc
     1621 tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac
     1681 acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg
     1741 ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg
     1801 gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag
     1861 gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg
     1921 gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg
     1981 aggctagcat cgattgatca acaagtttgt acaaaaaagt tggcatggac tataaggacc
     2041 acgacggaga ctacaaggat catgatattg attacaaaga cgatgacgat aagatggccc
     2101 caaagaagaa gcggaaggtc ggtatccacg gagtcccagc agccgacaag aagtacagca
     2161 tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg
     2221 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc
     2281 tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa
     2341 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca
     2401 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg
     2461 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg
     2521 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg
     2581 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc
     2641 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc
     2701 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg
     2761 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg
     2821 cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg
     2881 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga
     2941 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg
     3001 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga
     3061 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg
     3121 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt
     3181 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag
     3241 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg
     3301 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca
     3361 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg
     3421 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc
     3481 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca
     3541 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt
     3601 ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg
     3661 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag
     3721 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg
     3781 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg
     3841 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt
     3901 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc
     3961 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg
     4021 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag
     4081 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga
     4141 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg
     4201 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg
     4261 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc
     4321 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc
     4381 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga
     4441 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg
     4501 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc
     4561 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg
     4621 aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc
     4681 tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga
     4741 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc
     4801 tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg
     4861 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga
     4921 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg
     4981 acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc
     5041 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg
     5101 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg
     5161 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc
     5221 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca
     5281 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg
     5341 gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc
     5401 tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca
     5461 aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg
     5521 accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg
     5581 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca
     5641 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct
     5701 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg
     5761 aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg
     5821 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg
     5881 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg
     5941 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc
     6001 tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg
     6061 agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact
     6121 ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc
     6181 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag
     6241 gcgacaaaag gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aagaccccca
     6301 actttcttgt acaaagtggt tgatatcggt aagcctatcc ctaaccctct cctcggtctc
     6361 gattctacgt agtaatgaac tagtaccggt taagtcgaca atcaacgcgt taagtcgaca
     6421 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     6481 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     6541 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     6601 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     6661 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     6721 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     6781 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     6841 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     6901 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     6961 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt cgactttaag
     7021 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     7081 ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct
     7141 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     7201 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     7261 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtacgta
     7321 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     7381 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     7441 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     7501 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc
     7561 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     7621 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     7681 ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
     7741 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     7801 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     7861 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     7921 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     7981 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     8041 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     8101 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     8161 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     8221 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     8281 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     8341 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     8401 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     8461 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     8521 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     8581 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     8641 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     8701 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     8761 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     8821 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     8881 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     8941 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     9001 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     9061 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     9121 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     9181 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     9241 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     9301 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     9361 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     9421 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     9481 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     9541 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     9601 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     9661 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     9721 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     9781 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     9841 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     9901 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     9961 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
    10021 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
    10081 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
    10141 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
    10201 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
    10261 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
    10321 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
    10381 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
    10441 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
    10501 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca
    10561 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
    10621 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
    10681 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
    10741 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct
    10801 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
    10861 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
    10921 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
    10981 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
    11041 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
    11101 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
    11161 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
    11221 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
    11281 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
    11341 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
    11401 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
    11461 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
    11521 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
    11581 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
    11641 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
    11701 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
    11761 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
    11821 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
    11881 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
    11941 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
    12001 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
    12061 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
    12121 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
    12181 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
    12241 tcgtttcaga cccacctccc aaccccgagg ggacccatgc attgcatctc aattagtcag
    12301 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcgtc
    12361 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
    12421 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
    12481 aggccgcctc ggcc
//
