LOCUS       sequence-390148-        9220 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Human CASP9 Specific gRNA/Cas9 Complex and Reporter 
            Protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9220)
  AUTHORS   Han J, Goldstein LA, Hou W, Watkins SC, Rabinowich H
  TITLE     Involvement of CASP9 (caspase 9) in IGF2R/CI-MPR endosomal 
            transport.
  JOURNAL   Autophagy. 2021 Jun;17(6):1393-1409. doi: 
            10.1080/15548627.2020.1761742. Epub 2020 May 25.
  PUBMED    32397873
REFERENCE   2  (bases 1 to 9220)
  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"; doi: 
            "10.1080/15548627.2020.1761742"; journalName: "Autophagy"; date: 
            "2021-06"; volume: "17"; issue: "6"; pages: "1393-1409"
COMMENT     Sequence Label: pCRISPR Human CASP9 -1
FEATURES             Location/Qualifiers
     source          1..9220
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        55..434
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        435..638
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     589..609
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     635..659
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      733..742
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             742..783
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             787..807
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             832..4932
                     /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             832..4932
                     /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"
     RBS             4845..4853
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             4945..4965
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4987..5040
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5041..5706
                     /codon_start=1
                     /product="orange fluorescent protein from Cerianthus (Ip et
                     al., 2007)"
                     /label=OFP
                     /translation="MNLSKNVSVSVYMKGNVNNHEFEYDGEGGGDPYTGKYSMKMTLRG
                     QNSLPFSYDIITTAFQYGFRVFTKYPEGIVDYFKDSLPDAFQWNRRIVFEDGGVLNMSS
                     DITYKDNVLHGDVKAEGVNFPPNGPVMKNEIVMEEPTEETFTPKNGVLVGFCPKAYLLK
                     DGSYYYGNMTTFYRSKKSGQAPPGYHFVKHRLVKTNVGHGFKTVEQTEYATAHVSDLPK
                     "
     primer_bind     complement(5746..5765)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5790..5838
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6040..6468
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6127..6146)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6337..6358
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6488..6736
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6488..6508
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6659..6678
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6758..6833
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(6883..6899)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6883..6899)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6896..6918)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6907..6923
                     /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(6931..6937,6938..6955,6956..6961))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6976..6997
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7114..7131)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7285..7873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7365..7384)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8044..8835,8836..8904))
                     /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     8667..8686
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8905..9009)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9084..9103)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
       61 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      121 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      181 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      241 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      301 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      361 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      421 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      481 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      541 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      601 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg
      661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      721 tccggagcgg ccgccaccat gggcaagccc atccctaacc ccctgttggg gctggacagc
      781 accgctccca aaaagaaaag gaaggtgggc attcacggcg tgcctgcggc cgacaaaaag
      841 tacagcatcg gccttgatat cggcaccaat agcgtgggct gggccgttat cacagacgaa
      901 tacaaggtac ccagcaagaa gttcaaggtg ctggggaata cagacaggca ctctatcaag
      961 aaaaacctta tcggggctct gctgtttgac tcaggcgaga ccgccgaggc caccaggttg
     1021 aagaggaccg caaggcgaag gtacacccgg aggaagaaca ggatctgcta tctgcaggag
     1081 atcttcagca acgagatggc caaggtggac gacagcttct tccacaggct ggaggagagc
     1141 ttccttgtcg aggaggataa gaagcacgaa cgacacccca tcttcggcaa catagtcgac
     1201 gaggtcgctt atcacgagaa gtaccccacc atctaccacc tgcgaaagaa attggtggat
     1261 agcaccgata aagccgactt gcgacttatc tacttggctc tggcgcacat gattaagttc
     1321 aggggccact tcctgatcga gggcgacctt aaccccgaca acagtgacgt agacaaattg
     1381 ttcatccagc ttgtacagac ctataaccag ctgttcgagg aaaaccctat taacgccagc
     1441 ggggtggatg cgaaggccat acttagcgcc aggctgagca aaagcaggcg cttggagaac
     1501 ctgatagccc agctgcccgg tgaaaagaag aacggcctct tcggtaatct gattgccctg
     1561 agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggcagaaga tgccaagctg
     1621 cagttgagta aggacaccta tgacgacgac ttggacaatc tgctcgccca aatcggcgac
     1681 cagtacgctg acctgttcct cgccgccaag aacctttctg acgcaatcct gcttagcgat
     1741 atccttaggg tgaacacaga gatcaccaag gcccccctga gcgccagcat gatcaagagg
     1801 tacgacgagc accatcagga cctgaccctt ctgaaggccc tggtgaggca gcaactgccc
     1861 gagaagtaca aggagatctt tttcgaccag agcaagaacg gctacgccgg ctacatcgac
     1921 ggcggagcca gccaagagga gttctacaag ttcatcaagc ccatcctgga gaagatggat
     1981 ggcaccgagg agctgctggt gaagctgaac agggaagatt tgctccggaa gcagaggacc
     2041 tttgacaacg gtagcatccc ccaccagatc cacctgggcg agctgcacgc aatactgagg
     2101 cgacaggagg atttctaccc cttcctcaag gacaataggg agaaaatcga aaagattctg
     2161 accttcagga tcccctacta cgtgggccct cttgccaggg gcaacagccg attcgcttgg
     2221 atgacaagaa agagcgagga gaccatcacc ccctggaact tcgaggaagt ggtggacaaa
     2281 ggagcaagcg cgcagtcttt catcgaacgg atgaccaatt tcgacaaaaa cctgcctaac
     2341 gagaaggtgc tgcccaagca cagcctgctt tacgagtact tcaccgtgta caacgagctc
     2401 accaaggtga aatatgtgac cgagggcatg cgaaaacccg ctttcctgag cggcgagcag
     2461 aagaaggcca tcgtggacct gctgttcaag accaacagga aggtgaccgt gaagcagctg
     2521 aaggaggact acttcaagaa gatcgagtgc tttgatagcg tggaaataag cggcgtggag
     2581 gacaggttca acgccagcct gggcacctac cacgacttgt tgaagataat caaagacaag
     2641 gatttcctgg ataatgagga gaacgaggat atactcgagg acatcgtgct gactttgacc
     2701 ctgtttgagg accgagagat gattgaagaa aggctcaaaa cctacgccca cctgttcgac
     2761 gacaaagtga tgaaacaact gaagagacga agatacaccg gctggggcag actgtccagg
     2821 aagctcatca acggcattag ggacaagcag agcggcaaga ccatcctgga tttcctgaag
     2881 tccgacggct tcgccaaccg aaacttcatg cagctgattc acgatgacag cttgaccttc
     2941 aaggaggaca tccagaaggc ccaggttagc ggccagggcg actccctgca cgaacatatt
     3001 gcaaacctgg caggctcccc tgcgatcaag aagggcatac tgcagaccgt taaggttgtg
     3061 gacgaattgg tcaaggtcat gggcaggcac aagcccgaaa acatagttat agagatggcc
     3121 agagagaacc agaccaccca aaagggccag aagaacagcc gggagcgcat gaaaaggatc
     3181 gaggagggta tcaaggaact cggaagccag atcctcaaag agcaccccgt ggagaatacc
     3241 cagctccaga acgagaagct gtacctgtac tacctgcaga acggcaggga catgtacgtt
     3301 gaccaggagt tggacatcaa caggctttca gactatgacg tggatcacat agtgccccag
     3361 agctttctta aagacgatag catcgacaac aaggtcctga cccgctccga caaaaacagg
     3421 ggcaaaagcg acaacgtgcc aagcgaagag gtggttaaaa agatgaagaa ctactggagg
     3481 caactgctca acgcgaaatt gatcacccag agaaagttcg ataacctgac caaggccgag
     3541 aggggcggac tctccgaact tgacaaagcg ggcttcataa agaggcagct ggtcgagacc
     3601 cgacagatca cgaagcacgt ggcccaaatc ctcgacagca gaatgaatac caagtacgat
     3661 gagaatgaca aactcatcag ggaagtgaaa gtgattaccc tgaagagcaa gttggtgtcc
     3721 gactttcgca aagatttcca gttctacaag gtgagggaga tcaacaacta ccaccatgcc
     3781 cacgacgcat acctgaacgc cgtggtcggc accgccctga ttaagaagta tccaaagctg
     3841 gagtccgaat ttgtctacgg cgactacaaa gtttacgatg tgaggaagat gatcgctaag
     3901 agcgaacagg agatcggcaa ggccaccgct aagtatttct tctacagcaa catcatgaac
     3961 tttttcaaga ccgagatcac acttgccaac ggcgaaatca ggaagaggcc gcttatcgag
     4021 accaacggtg agaccggcga gatcgtgtgg gacaagggca gggacttcgc caccgtgagg
     4081 aaagtcctga gcatgcccca ggtgaatatt gtgaaaaaaa ctgaggtgca gacaggcggc
     4141 tttagcaagg aatccatcct gcccaagagg aacagcgaca agctgatcgc ccggaagaag
     4201 gactgggacc ctaagaagta tggaggcttc gacagcccca ccgtagccta cagcgtgctg
     4261 gtggtcgcga aggtagagaa ggggaagagc aagaaactga agagcgtgaa ggagctgctc
     4321 ggcataacca tcatggagag gtccagcttt gagaagaacc ccattgactt tttggaagcc
     4381 aagggctaca aagaggtcaa aaaggacctg atcatcaaac tccccaagta ctccctgttt
     4441 gaattggaga acggcagaaa gaggatgctg gcgagcgctg gggaactgca aaagggcaac
     4501 gaactggcgc tgcccagcaa gtacgtgaat tttctgtacc tggcgtccca ctacgaaaag
     4561 ctgaaaggca gccccgagga caacgagcag aagcagctgt tcgtggagca gcacaagcat
     4621 tacctggacg agataatcga gcaaatcagc gagttcagca agagggtgat tctggccgac
     4681 gcgaacctgg ataaggtcct cagcgcctac aacaagcacc gagacaaacc catcagggag
     4741 caggccgaga atatcataca cctgttcacc ctgacaaatc tgggcgcacc tgcggcattc
     4801 aaatacttcg ataccaccat cgacaggaaa aggtacacta gcactaagga ggtgctggat
     4861 gccaccttga tccaccagtc cattaccggc ctgtatgaga ccaggatcga cctgagccag
     4921 cttggaggcg actctagggc ggacccaaaa aagaaaagga aggtggaatt ctctagaggc
     4981 agtggagagg gcagaggaag tctgctaaca tgcggtgacg tcgaggagaa tcctggccca
     5041 atgaacctga gcaaaaacgt gagcgtgagc gtgtatatga aggggaacgt caacaatcat
     5101 gagtttgagt acgacgggga aggtggtggt gatccttata caggtaaata ttccatgaag
     5161 atgacgctac gtggtcaaaa ttccctaccc ttttcctatg atatcattac cacggcattt
     5221 cagtatggtt tccgcgtatt tacaaaatac cctgagggaa ttgttgacta ttttaaggac
     5281 tcgcttcccg acgcattcca gtggaacaga cgaattgtgt ttgaagatgg tggagtacta
     5341 aacatgagca gtgatatcac atataaagat aatgttctgc atggtgacgt caaggctgag
     5401 ggagtgaact tcccgccgaa tgggccagtg atgaagaatg aaattgtgat ggaggaaccg
     5461 actgaagaaa catttactcc aaaaaacggg gttcttgttg gcttttgtcc caaagcgtac
     5521 ttacttaaag acggttccta ttactatgga aatatgacaa cattttacag atccaagaaa
     5581 tctggccagg cacctcctgg gtatcacttt gttaagcatc gtctcgtcaa gaccaatgtg
     5641 ggacatggat ttaagacggt tgagcagact gaatatgcca ctgctcatgt cagtgatctt
     5701 cccaagttcg aagcttgata atgagtttaa acgggggagg ctaactgaaa cacggaagga
     5761 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     5821 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     5881 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     5941 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     6001 tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     6061 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     6121 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     6181 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     6241 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     6301 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     6361 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     6421 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttaaggtcgg
     6481 gcaggaagag ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt
     6541 tagagagata attagaatta atttgactgt aaacacaaag atattagtac aaaatacgtg
     6601 acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga
     6661 ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg
     6721 gaaaggacga aacaccgaca cccagaccag tggacatgtt ttagagctag aaatagcaag
     6781 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttc
     6841 tagtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     6901 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     6961 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     7021 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     7081 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     7141 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     7201 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     7261 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     7321 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7381 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7441 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7501 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7561 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7621 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7681 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     7741 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7801 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7861 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7921 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     7981 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     8041 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     8101 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     8161 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     8221 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     8281 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     8341 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     8401 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     8461 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8521 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8581 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8641 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8701 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8761 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8821 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8881 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     8941 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     9001 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     9061 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     9121 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     9181 acaaggcaag gcttgaccga caattgcatg aagaatctgc
//
