LOCUS       sequence-366611-       11211 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral Cas9 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11211)
  TITLE     Klusmann Lab plasmids 2022
REFERENCE   2  (bases 1 to 11211)
  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"
COMMENT     Sequence Label: pLKO5d.SFFV.Cas9.P2A.BSD
FEATURES             Location/Qualifiers
     source          1..11211
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    41..158
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        216..623
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      623..632
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             632..655
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             662..682
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             707..4807
                     /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             707..4807
                     /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             4808..4855
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4871..4927
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4928..5323
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    5357..5945
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5410..5430)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5828..5839)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6017..6250
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6328..6449
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6365..6384)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6419..6438
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6489..6624
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6644..6663)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6645..6663)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6673..6690)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6673..6689)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6682..6704)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6831..7286
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6918..6937)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7128..7149
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7312..7416
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7417..7485,7486..8277)
                     /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(7635..7654)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8448..9036
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8937..8956
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9190..9207
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9324..9345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9360..9365,9366..9383,9384..9390)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9398..9414
                     /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     9403..9425
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9422..9438
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9422..9438
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9457..9477
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9459..9477
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9505..9731
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             9732..9912
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9959..10084
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10577..10810
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10995..11039
                     /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"
ORIGIN
        1 aattctagat cttgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
       61 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
      121 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
      181 gagatccact ttggcgccgg ctcgagcgag ctgcagtaac gccattttgc aaggcatgga
      241 aaaataccaa accaagaata gagaagttca gatcaagggc gggtacatga aaatagctaa
      301 cgttgggcca aacaggatat ctgcggtgag cagtttcggc cccggcccgg ggccaagaac
      361 agatggtcac cgcagtttcg gccccggccc gaggccaaga acagatggtc cccagatatg
      421 gcccaaccct cagcagtttc ttaagaccca tcagatgttt ccaggctccc ccaaggacct
      481 gaaatgaccc tgcgccttat ttgaattaac caatcagcct gcttctcgct tctgttcgcg
      541 cgcttctgct tcccgagctc tataaaagag ctcacaaccc ctcactcggc gcgccagtcc
      601 tccgacagac tgagtcggat ccgccaccat ggattacaaa gacgatgacg ataagatggc
      661 cccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccgaca agaagtacag
      721 catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa
      781 ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa
      841 cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag
      901 aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt
      961 cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag agtccttcct
     1021 ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt
     1081 ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac
     1141 cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg
     1201 ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat
     1261 ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt
     1321 ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat
     1381 cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct
     1441 gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct
     1501 gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta
     1561 cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct
     1621 gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga
     1681 cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa
     1741 gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg
     1801 agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac
     1861 cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga
     1921 caacggcagc atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca
     1981 ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt
     2041 ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac
     2101 cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc
     2161 ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa
     2221 ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa
     2281 agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa
     2341 ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga
     2401 ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg
     2461 gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt
     2521 cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt
     2581 tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa
     2641 agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct
     2701 gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga
     2761 cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga
     2821 ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa
     2881 tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga
     2941 gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga
     3001 gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga
     3061 gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct
     3121 gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca
     3181 ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt
     3241 tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa
     3301 gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct
     3361 gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg
     3421 cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca
     3481 gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa
     3541 tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt
     3601 ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga
     3661 cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag
     3721 cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga
     3781 gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt
     3841 caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa
     3901 cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt
     3961 gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag
     4021 caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg
     4081 ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt
     4141 ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat
     4201 caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg
     4261 ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct
     4321 ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact
     4381 ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa
     4441 gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct
     4501 ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa
     4561 tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc
     4621 cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta
     4681 ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac
     4741 cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg
     4801 aggcgacaag cgtcctgctg ctactaagaa agctggtcaa gctaagaaaa agaaagctag
     4861 cggcagcggc gccaccaact tcagcctgct gaagcaggcc ggcgacgtgg aggagaaccc
     4921 cggccccatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc
     4981 tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag
     5041 cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga
     5101 actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc
     5161 gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct
     5221 cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt
     5281 tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggcctgcagc tgcagtagta
     5341 aacgcgttaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     5401 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     5461 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     5521 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     5581 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     5641 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     5701 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     5761 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     5821 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     5881 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     5941 cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     6001 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
     6061 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     6121 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     6181 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     6241 atctctagca gtacgtatag tagttcatgt catcttatta ttcagtattt ataacttgca
     6301 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     6361 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6421 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     6481 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     6541 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     6601 agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta
     6661 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     6721 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     6781 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     6841 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     6901 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     6961 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     7021 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     7081 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     7141 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     7201 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     7261 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     7321 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     7381 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     7441 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     7501 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     7561 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     7621 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     7681 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     7741 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     7801 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     7861 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     7921 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     7981 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     8041 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     8101 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     8161 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     8221 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     8281 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     8341 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     8401 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     8461 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     8521 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     8581 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     8641 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     8701 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     8761 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     8821 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     8881 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     8941 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     9001 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     9061 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     9121 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     9181 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     9241 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     9301 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     9361 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     9421 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
     9481 caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca
     9541 tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat
     9601 tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat
     9661 ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc
     9721 gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     9781 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     9841 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     9901 aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct
     9961 ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact
    10021 ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag
    10081 cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg
    10141 gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt
    10201 cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct
    10261 acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac
    10321 cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat
    10381 agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac
    10441 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
    10501 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
    10561 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
    10621 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
    10681 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
    10741 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
    10801 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
    10861 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
    10921 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
    10981 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
    11041 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
    11101 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
    11161 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga c
//
