LOCUS       sequence-91238-e        8302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9/gRNA Entry expression plasmid .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8302)
  AUTHORS   Gonzalez F, Zhu Z, Shi ZD, Lelli K, Verma N, Li QV, Huangfu D
  TITLE     An iCRISPR Platform for Rapid, Multiplexable, and Inducible Genome 
            Editing in Human Pluripotent Stem Cells.
  JOURNAL   Cell Stem Cell. 2014 Jun 11. pii: S1934-5909(14)00205-7. doi: 
            10.1016/j.stem.2014.05.018.
  PUBMED    24931489
REFERENCE   2  (bases 1 to 8302)
  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: "Cell Stem 
            Cell. 2014 Jun 11. pii: S1934-5909(14)00205-7. doi: 
            10.1016/j.stem.2014.05.018."
COMMENT     Sequence Label: piCRg Entry
FEATURES             Location/Qualifiers
     source          1..8302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..69)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    155..295
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     210..229
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(310..327)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(481..1069)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(561..580)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1296..2090)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(1296..2090)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1407..1426)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2017..2036
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2278..2295
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2279..2295
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2302..2321
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2302..2320
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2337..2847
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2766..2784
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     protein_bind    2858..2957
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      2982..2991
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2991..3056
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3063..3083
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3108..7208
                     /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             3108..7208
                     /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             7209..7256
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(7284..7301)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7290..7497
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        7506..7754
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7506..7526
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7677..7696
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7773..7848
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(7966..8065)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(8114..8132)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(8115..8132)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(8150..8166)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8150..8166)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8163..8185)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8174..8190
                     /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(8198..8204,8205..8222,8223..8228))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8243..8264
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cctgtcgtgc cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
       61 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
      121 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
      181 atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
      241 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc
      301 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
      361 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
      421 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
      481 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
      541 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      601 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      661 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      721 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      781 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      841 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      901 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
      961 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1021 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     1081 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     1141 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     1201 atttccccga aaagtgccac ctgtatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     1261 aaataccgca tcaggaaatt gtaagcgtta ataattcaga agaactcgtc aagaaggcga
     1321 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
     1381 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
     1441 cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
     1501 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
     1561 ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
     1621 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
     1681 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
     1741 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
     1801 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
     1861 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc
     1921 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
     1981 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
     2041 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
     2101 cctcatcctg tctcttgatc agagcttgat cccctgcgcc atcagatcct tggcggcgag
     2161 aaagccatcc agtttacttt gcagggcttc ccaaccttac cagagggcgc cccagctggc
     2221 aattccggtt cgcttgctgt ccataaaacc gcccagtcta gctatcgcca tgtaagttgt
     2281 aaaacgacgg ccagtgaatt gtaatacgac tcactatagg gcgaattgaa atccacgggg
     2341 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
     2401 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
     2461 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
     2521 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
     2581 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
     2641 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
     2701 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
     2761 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
     2821 gaccgaatca ccgacctctc tccccaggga tcccccccaa ataatgattt tattttgact
     2881 gatagtgacc tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt
     2941 gtacaaaaaa gcaggctccg cggttttcag gttggaccgg tgccaccatg gactataagg
     3001 accacgacgg agactacaag gatcatgata ttgattacaa agacgatgac gataagatgg
     3061 ccccaaagaa gaagcggaag gtcggtatcc acggagtccc agcagccgac aagaagtaca
     3121 gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca
     3181 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga
     3241 acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga
     3301 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct
     3361 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc
     3421 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg
     3481 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca
     3541 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg
     3601 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca
     3661 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg
     3721 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga
     3781 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc
     3841 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc
     3901 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt
     3961 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc
     4021 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg
     4081 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga
     4141 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg
     4201 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca
     4261 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg
     4321 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc
     4381 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct
     4441 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga
     4501 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg
     4561 cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga
     4621 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca
     4681 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa
     4741 aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag
     4801 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc
     4861 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact
     4921 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt
     4981 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca
     5041 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc
     5101 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg
     5161 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag
     5221 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca
     5281 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg
     5341 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag
     5401 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag
     5461 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc
     5521 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc
     5581 aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct
     5641 ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca
     5701 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc
     5761 tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag
     5821 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc
     5881 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga
     5941 atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
     6001 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg
     6061 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa
     6121 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg
     6181 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt
     6241 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa
     6301 acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag
     6361 tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca
     6421 gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact
     6481 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg
     6541 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga
     6601 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg
     6661 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc
     6721 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac
     6781 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga
     6841 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc
     6901 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta
     6961 atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg
     7021 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt
     7081 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca
     7141 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg
     7201 gaggcgacaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag aaaaagtaag
     7261 aattcctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     7321 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     7381 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     7441 ggtggggcag gacagcaagg gggaggattg ggaagagaat agcaggcatg ctggggagcg
     7501 gccgcgaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
     7561 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
     7621 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
     7681 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
     7741 aaggacgaaa caccgggtct tcgagaagac ctgttttaga gctagaaata gcaagttaaa
     7801 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttttgtttta
     7861 gagctagaaa tagcaagtta aaataaggct agtccgtttt tagcgcgtgc gccaattctg
     7921 cagacaaatg gctctagagg tacgaattca agggtgggcg cgccgaccca gctttcttgt
     7981 acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta
     8041 tcagtcaaaa taaaatcatt atttgccatc cagatccgag ctcggtacca agcttgatgc
     8101 atagcttgag tattctatag tgtcacctaa atagcttggc gtaatcatgg tcatagctgt
     8161 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     8221 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     8281 tgcccgcttt ccagtcggga aa
//
