LOCUS       sequence-181653-       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"
     primer_bind     complement(30..47)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(201..789)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(281..300)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(960..1751,1752..1820))
                     /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     1583..1602
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1821..1925)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1951..2406)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2088..2109)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2300..2319
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2533..2555
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2547..2564
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2548..2564
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2574..2593
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2574..2592
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      2613..2748
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2788..2909
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2799..2818)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2853..2872
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             2987..3220
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    3292..3880
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3398..3409
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3807..3827
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3931..3972)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    3980..4004
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(4007..4054)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(4055..8155)
                     /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             complement(4055..8155)
                     /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             complement(8180..8200)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8207..8272)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(8275..8299)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(8319..9497)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          8328..9266
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(8351..8371)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    9635..9752
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(9826..10221)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(10243..10439)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        complement(10251..10461)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      10257..10392
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             complement(10692..10736)
                     /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"
     misc_feature    10921..11154
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    11647..11772
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             11819..11999
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(12000..12226)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(12254..12274)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(12254..12272)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(12293..12309)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12293..12309)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12306..12328)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12317..12333
                     /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)."
     promoter        complement(join(12341..12347,12348..12365,12366..12371))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12386..12407
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
       61 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      121 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      181 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      241 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      301 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      361 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      421 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      481 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      541 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      601 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      661 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      721 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      781 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
      841 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
      901 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
      961 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1021 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     1081 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     1141 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     1201 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     1261 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     1321 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     1381 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     1441 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     1501 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     1561 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     1621 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     1681 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     1741 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     1801 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     1861 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     1921 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     1981 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     2041 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     2101 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     2161 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     2221 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     2281 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     2341 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     2401 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     2461 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     2521 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg
     2581 actcactata gggcgaattg ggtacgtccc taggcctcca aaaaagcctc ctcactactt
     2641 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca
     2701 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
     2761 gggcgggata gctagagcca gacatgataa gatacattga tgagtttgga caaaccacaa
     2821 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     2881 taaccattat aagctgcaat aaacaagttc ctctcactct ctgatattca tttctttgca
     2941 agttataaat actgaataat aagatgacat gaactactat acgtactgct agagattttc
     3001 cacactgact aaaagggtct gagggatctc tagttaccag agtcacacaa cagacgggca
     3061 cacactactt gaagcactca aggcaagctt tattgaggct taagcagtgg gttccctagt
     3121 tagccagaga gctcccaggc tcagatctgg tctaaccaga gagacccagt acaagcaaaa
     3181 agcagatctt gtcttcgttg ggagtgaatt agcccttcca gtcccccctt ttcttttaaa
     3241 aagtggctaa gatctacagc tgccttgtaa gtcattggtc ttaaagtcga cgcggggagg
     3301 cggcccaaag ggagatccga ctcgtctgag ggcgaaggcg aagacgcgga agaggccgca
     3361 gagccggcag caggccgcgg gaaggaaggt ccgctggatt gagggccgaa gggacgtagc
     3421 agaaggacgt cccgcgcaga atccaggtgg caacacaggc gagcagccaa ggaaaggacg
     3481 atgatttccc cgacaacacc acggaattgt cagtgcccaa cagccgagcc cctgtccagc
     3541 agcgggcaag gcaggcggcg atgagttccg ccgtggcaat agggaggggg aaagcgaaag
     3601 tcccggaaag gagctgacag gtggtggcaa tgccccaacc agtgggggtt gcgtcagcaa
     3661 acacagtgca caccacgcca cgttgcctga caacgggcca caactcctca taaagagaca
     3721 gcaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat
     3781 acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac
     3841 cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt gtcgacttaa cgcgttgatt
     3901 gtcgacttaa ccggtactag ttcattacta cgtagaatcg agaccgagga gagggttagg
     3961 gataggctta ccgatatcaa ccactttgta caagaaagtt gggggtcttt ttcttttttg
     4021 cctggccggc ctttttcgtg gccgccggcc ttttgtcgcc tcccagctga gacaggtcga
     4081 tccgtgtctc gtacaggccg gtgatgctct ggtggatcag ggtggcgtcc agcacctctt
     4141 tggtgctggt gtacctcttc cggtcgatgg tggtgtcaaa gtacttgaag gcggcagggg
     4201 ctcccagatt ggtcagggta aacaggtgga tgatattctc ggcctgctct ctgatgggct
     4261 tatcccggtg cttgttgtag gcggacagca ctttgtccag attagcgtcg gccaggatca
     4321 ctctcttgga gaactcgctg atctgctcga tgatctcgtc caggtagtgc ttgtgctgtt
     4381 ccacaaacag ctgtttctgc tcattatcct cgggggagcc cttcagcttc tcatagtggc
     4441 tggccaggta caggaagttc acatatttgg agggcagggc cagttcgttt cccttctgca
     4501 gttcgccggc agaggccagc attctcttcc ggccgttttc cagctcgaac agggagtact
     4561 taggcagctt gatgatcagg tcctttttca cttctttgta gcccttggct tccagaaagt
     4621 cgatgggatt cttctcgaag ctgcttcttt ccatgatggt gatccccagc agctctttca
     4681 cactcttcag tttcttggac ttgccctttt ccactttggc caccaccagc acagaatagg
     4741 ccacggtggg gctgtcgaag ccgccgtact tcttagggtc ccagtccttc tttctggcga
     4801 tcagcttatc gctgttcctc ttgggcagga tagactcttt gctgaagccg cctgtctgca
     4861 cctcggtctt tttcacgata ttcacttggg gcatgctcag cactttccgc acggtggcaa
     4921 aatcccggcc cttatcccac acgatctccc cggtttcgcc gtttgtctcg atcagaggcc
     4981 gcttccggat ctcgccgttg gccagggtaa tctcggtctt gaaaaagttc atgatgttgc
     5041 tgtagaagaa gtacttggcg gtagccttgc cgatttcctg ctcgctcttg gcgatcatct
     5101 tccgcacgtc gtacaccttg tagtcgccgt acacgaactc gctttccagc ttagggtact
     5161 ttttgatcag ggcggttccc acgacggcgt tcaggtaggc gtcgtgggcg tggtggtagt
     5221 tgttgatctc gcgcactttg taaaactgga aatccttccg gaaatcggac accagcttgg
     5281 acttcagggt gatcactttc acttcccgga tcagcttgtc attctcgtcg tacttagtgt
     5341 tcatccggga gtccaggatc tgtgccacgt gctttgtgat ctgccgggtt tccaccagct
     5401 gtctcttgat gaagccggcc ttatccagtt cgctcaggcc gcctctctcg gccttggtca
     5461 gattgtcgaa ctttctctgg gtaatcagct tggcgttcag cagctgccgc cagtagttct
     5521 tcatcttctt cacgacctct tcggagggca cgttgtcgct cttgccccgg ttcttgtcgc
     5581 ttctggtcag caccttgttg tcgatggagt cgtccttcag aaagctctga ggcacgatat
     5641 ggtccacatc gtagtcggac agccggttga tgtccagttc ctggtccacg tacatatccc
     5701 gcccattctg caggtagtac aggtacagct tctcgttctg cagctgggtg ttttccacgg
     5761 ggtgttcttt caggatctgg ctgcccagct ctttgatgcc ctcttcgatc cgcttcattc
     5821 tctcgcggct gttcttctgt cccttctggg tggtctggtt ctctctggcc atttcgatca
     5881 cgatgttctc gggcttgtgc cggcccatca ctttcacgag ctcgtccacc accttcactg
     5941 tctgcaggat gcccttctta atggcggggc tgccggccag attggcaatg tgctcgtgca
     6001 ggctatcgcc ctggccggac acctgggctt tctggatgtc ctctttaaag gtcaggctgt
     6061 cgtcgtggat cagctgcatg aagtttctgt tggcgaagcc gtcggacttc aggaaatcca
     6121 ggattgtctt gccggactgc ttgtcccgga tgccgttgat cagcttccgg ctcagcctgc
     6181 cccagccggt gtatctccgc cgcttcagct gcttcatcac tttgtcgtcg aacaggtggg
     6241 cataggtttt cagccgttcc tcgatcatct ctctgtcctc aaacagtgtc agggtcagca
     6301 cgatatcttc cagaatgtcc tcgttttcct cattgtccag gaagtccttg tccttgataa
     6361 ttttcagcag atcgtggtat gtgcccaggg aggcgttgaa ccgatcttcc acgccggaga
     6421 tttccacgga gtcgaagcac tcgattttct tgaagtagtc ctctttcagc tgcttcacgg
     6481 tcactttccg gttggtcttg aacagcaggt ccacgatggc ctttttctgc tcgccgctca
     6541 ggaaggcggg ctttctcatt ccctcggtca cgtatttcac tttggtcagc tcgttataca
     6601 cggtgaagta ctcgtacagc aggctgtgct tgggcagcac cttctcgttg ggcaggttct
     6661 tatcgaagtt ggtcatccgc tcgatgaagc tctgggcgga agcgcccttg tccaccactt
     6721 cctcgaagtt ccagggggtg atggtttcct cgctctttct ggtcatccag gcgaatctgc
     6781 tgtttcccct ggccagaggg cccacgtagt aggggatgcg gaaggtcagg atcttctcga
     6841 tcttttcccg gttgtccttc aggaatgggt aaaaatcttc ctgccgccgc agaatggcgt
     6901 gcagctctcc caggtggatc tggtggggga tgctgccgtt gtcgaaggtc cgctgcttcc
     6961 gcagcaggtc ctctctgttc agcttcacga gcagttcctc ggtgccgtcc atcttttcca
     7021 ggatgggctt gatgaacttg tagaactctt cctggctggc tccgccgtca atgtagccgg
     7081 cgtagccgtt cttgctctgg tcgaagaaaa tctctttgta cttctcaggc agctgctgcc
     7141 gcacgagagc tttcagcagg gtcaggtcct ggtggtgctc gtcgtatctc ttgatcatag
     7201 aggcgctcag gggggccttg gtgatctcgg tgttcactct caggatgtcg ctcagcagga
     7261 tggcgtcgga caggttcttg gcggccagaa acaggtcggc gtactggtcg ccgatctggg
     7321 ccagcaggtt gtccaggtcg tcgtcgtagg tgtccttgct cagctgcagt ttggcatcct
     7381 cggccaggtc gaagttgctc ttgaagttgg gggtcaggcc caggctcagg gcaatcaggt
     7441 ttccgaacag gccattcttc ttctcgccgg gcagctgggc gatcagattt tccagccgtc
     7501 tgctcttgct cagtctggca gacaggatgg ccttggcgtc cacgccgctg gcgttgatgg
     7561 ggttttcctc gaacagctgg ttgtaggtct gcaccagctg gatgaacagc ttgtccacgt
     7621 cgctgttgtc ggggttcagg tcgccctcga tcaggaagtg gccccggaac ttgatcatgt
     7681 gggccagggc cagatagatc agccgcaggt cggccttgtc ggtgctgtcc accagtttct
     7741 ttctcaggtg gtagatggtg gggtacttct cgtggtaggc cacctcgtcc acgatgttgc
     7801 cgaagatggg gtgccgctcg tgcttcttat cctcttccac caggaaggac tcttccagtc
     7861 tgtggaagaa gctgtcgtcc accttggcca tctcgttgct gaagatctct tgcagatagc
     7921 agatccggtt cttccgtctg gtgtatcttc ttctggcggt tctcttcagc cgggtggcct
     7981 cggctgtttc gccgctgtcg aacagcaggg ctccgatcag gttcttcttg atgctgtgcc
     8041 ggtcggtgtt gcccagcacc ttgaatttct tgctgggcac cttgtactcg tcggtgatca
     8101 cggcccagcc cacagagttg gtgccgatgt ccaggccgat gctgtacttc ttgtcggctg
     8161 ctgggactcc gtggataccg accttccgct tcttctttgg ggccatctta tcgtcatcgt
     8221 ctttgtaatc aatatcatga tccttgtagt ctccgtcgtg gtccttatag tccatgccaa
     8281 cttttttgta caaacttgtt gatcaatcga tgctagcctc acgacacctg aaatggaaga
     8341 aaaaaacttt gaaccactgt ctgaggcttg agaatgaacc aagatccaaa ctcaaaaagg
     8401 gcaaattcca aggagaatta catcaagtgc caagctggcc taacttcagt ctccacccac
     8461 tcagtgtggg gaaactccat cgcataaaac ccctcccccc aacctaaaga cgacgtactc
     8521 caaaagctcg agaactaatc gaggtgcctg gacggcgccc ggtactccgt ggagtcacat
     8581 gaagcgacgg ctgaggacgg aaaggccctt ttcctttgtg tgggtgactc acccgcccgc
     8641 tctcccgagc gccgcgtcct ccattttgag ctccctgcag cagggccggg aagcggccat
     8701 ctttccgctc acgcaactgg tgccgaccgg gccagccttg ccgcccaggg cggggcgata
     8761 cacggcggcg cgaggccagg caccagagca ggccggccag cttgagacta cccccgtccg
     8821 attctcggtg gccgcgctcg caggccccgc ctcgccgaac atgtgcgctg ggacgcacgg
     8881 gccccgtcgc cgcccgcggc cccaaaaacc gaaataccag tgtgcagatc ttggcccgca
     8941 tttacaagac tatcttgcca gaaaaaaagc gtcgcagcag gtcatcaaaa attttaaatg
     9001 gctagagact tatcgaaagc agcgagacag gcgcgaaggt gccaccagat tcgcacgcgg
     9061 cggccccagc gcccaggcca ggcctcaact caagcacgag gcgaaggggc tccttaagcg
     9121 caaggcctcg aactctccca cccacttcca acccgaagct cgggatcaag aatcacgtac
     9181 tgcagccagg tggaagtaat tcaaggcacg caagggccat aacccgtaaa gaggccaggc
     9241 ccgcgggaac cacacacggc acttacctgt gttctggcgg caaacccgtt gcgaaaaaga
     9301 acgttcacgg cgactactgc acttatatac ggttctcccc caccctcggg aaaaaggcgg
     9361 agccagtaca cgacatcact ttcccagttt accccgcgcc accttctcta ggcaccggtt
     9421 caattgccga cccctccccc caacttctcg gggactgtgg gcgatgtgcg ctctgcccac
     9481 tgacgggcac cggagccaat tcccactcct ttcaagacct agaaggtcca ttagctgcaa
     9541 agattcctct ctgtttaaaa ctttatccat ctttgcacct gcaggcctcg agccggcgcc
     9601 aaagtggatc tctgctgtcc ctgtaataaa cccgaaaatt ttgaattttt gtaatttgtt
     9661 tttgtaattc tttagtttgt atgtctgttg ctattatgtc tactattctt tcccctgcac
     9721 tgtacccccc aatcccccct tttcttttaa aattgtggat gaatactgcc atttgtctca
     9781 agatctagaa ttcgttaacg gcgcgcctta ctactgcagc tgcaggccct cccacacata
     9841 accagagggc agcaattcac gaatcccaac tgccgtcggc tgtccatcac tgtccttcac
     9901 tatggctttg atcccaggat gcagatcgag aagcacctgt cggcaccgtc cgcaggggct
     9961 caagatgccc ctgttctcat ttccgatcgc gacgatacaa gtcaggttgc cagctgccgc
    10021 agcagcagca gtgcccagca ccacgagttc tgcacaaggt cccccagtaa aatgatatac
    10081 attgacacca gtgaagatgc ggccgtcgct agagagagct gcgctggcga cgctgtagtc
    10141 ttcagagatg gggatgctgt tgattgtagc cgttgctctt tcaatgaggg tggattcttc
    10201 ttgagacaaa ggcttggcca tggtggtacc tctagaaagc tttttgcaaa agcctaggcc
    10261 tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc
    10321 tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt
    10381 taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat tgagacgcat
    10441 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta
    10501 attgagatgc aatgcatggg tcccctcggg gttgggaggt gggtctgaaa cgataatggt
    10561 gaatatccct gcctaactct attcactata gaaagtacag caaaaactat tcttaaacct
    10621 accaagcctc ctactatcat tatgaataat tttatatacc acagccaatt tgttatgtta
    10681 aaccaattcc acaaacttgc ccatttatct aattccaata attcttgttc attcttttct
    10741 tgctggtttt gcgattcttc aattaaggag tgtattaagc ttgtgtaatt gttaatttct
    10801 ctgtcccact ccatccaggt cgtgtgattc caaatctgtt ccagagattt attactccaa
    10861 ctagcattcc aaggcacagc agtggtgcaa atgagttttc cagagcaacc ccaaatcccc
    10921 aggagctgtt gatcctttag gtatctttcc acagccagga ttcttgcctg gagctgcttg
    10981 atgccccaga ctgtgagttg caacagatgc tgttgcgcct caatagccct cagcaaattg
    11041 ttctgctgct gcactatacc agacaataat tgtctggcct gtaccgtcag cgtcattgac
    11101 gctgcgccca tagtgcttcc tgctgctccc aagaacccaa ggaacaaagc tcctattccc
    11161 actgctcttt tttctctctg caccactctt ctctttgcct tggtgggtgc tactcctaat
    11221 ggttcaattt ttactacttt atatttatat aattcacttc tccaattgtc cctcatatct
    11281 cctcctccag gtctgaagat cagcggccgc ttgctgtgcg gtggtcttac ttttgttttg
    11341 ctcttcctct atcttgtcta aagcttcctt ggtgtctttt atctctatcc tttgatgcac
    11401 acaatagagg gttgctactg tattatataa tgatctaagt tcttctgatc ctgtctgaag
    11461 ggatggttgt agctgtccca gtatttgtct acagccttct gatgtttcta acaggccagg
    11521 attaactgcg aatcgttcta gctccctgct tgcccatact atatgtttta atttatattt
    11581 tttctttccc cctggcctta accgaatttt ttcccatcgc gatctaattc tcccccgctt
    11641 aatactgacg ctctcgcacc catctctctc cttctagcct ccgctagtca aaatttttgg
    11701 cgtactcacc agtcgccgcc cctcgcctct tgccgtgcgc gcttcagcaa gccgagtcct
    11761 gcgtcgagag agctcctctg gtttcccttt cgctttcaag tccctgttcg ggcgccactg
    11821 ctagagattt tccacactga ctaaaagggt ctgagggatc tctagttacc agagtcacac
    11881 aacagacggg cacacactac ttgaagcact caaggcaagc tttattgagg cttaagcagt
    11941 gggttcccta gttagccaga gagctcccag gctcagatct ggtctaacca gagagacccg
    12001 tttatgtatc gagctaggca cttaaataca atatctctgc aatgcggcaa ttcagtggtt
    12061 cgtccaatcc atgtcagacc cgtctgttgc cttcctaata aggcacgatc gtaccacctt
    12121 acttccacca atcggcatgc acggtgcttt ttctctcctt gtaaggcatg ttgctaactc
    12181 atcgttacca tgttgcaaga ctacaagagt attgcataag actacattaa gcttgcagct
    12241 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
    12301 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
    12361 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
    12421 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
    12481 gcgcggggag aggc
//
