LOCUS       sequence-105977-        8986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian gRNA expression vector also expressing CIB1-dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8986)
  AUTHORS   Chen Z, Li W, Qiu F, Huang Q, Jiang Z, Ye J, Cheng P, Low C, Guo Y, 
            Yi X, Chen W, Yu Y, Han Y, Wu J, Jin S, Kong D, Huang J
  TITLE     Aspirin cooperates with p300 to activate the acetylation of H3K9 and
            promote FasL-mediated apoptosis of cancer stem-like cells in 
            colorectal cancer.
  JOURNAL   Theranostics. 2018 Aug 7;8(16):4447-4461. doi: 10.7150/thno.24284. 
            eCollection 2018.
  PUBMED    30214631
REFERENCE   2  (bases 1 to 8986)
  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: 
            "10.7150/thno.24284"; journalName: "Theranostics"; date: "2018-08-7-
            7"; volume: "8"; issue: "16"; pages: "4447-4461"
COMMENT     Sequence Label: px330-CIB1-dCas9
FEATURES             Location/Qualifiers
     source          1..8986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     286..306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     377..396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      403..412
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             412..477
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             484..504
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1507..1530,1531..1533,1534..4020,4021..4023,
                     4024..5610)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(1507..4905,4906..4908,4909..5505,5506,5507..5509,
                     5510..5511,5512..5514,5515..5607)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5616..5633
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5656..5673)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5662..5886
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5957..5973)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5957..5973)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5970..5992)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5981..5997
                     /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(6005..6011,6012..6029,6030..6035))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6050..6071
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6188..6205)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6359..6947)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6439..6458)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7118..7909,7910..7978))
                     /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     7741..7760
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7979..8083)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8162..8181)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        8352..8600
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8352..8372
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8523..8542
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8619..8694
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        join(8814..8986,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat ggactacaaa
      421 gaccatgacg gtgattataa agatcatgac atcgattaca aggatgacga tgacaagatg
      481 gcccccaaga agaagaggaa ggtgggcatt caccgcgggg tacctggagg ttctggatcc
      541 aacggcgcga ttggtgggga tttgctgctt aactttcccg acatgtccgt gttggaacgt
      601 cagcgcgcac atttgaagta tcttaacccc accttcgact ccccgttggc cgggttcttt
      661 gcggactcat ctatgattac gggaggggaa atggacagct acctctcaac ggccggattg
      721 aatcttccga tgatgtatgg agaaaccact gtagaaggcg actcgcgact ctcgatttcg
      781 cctgaaacga cgctgggaac agggaacttc aaggcggcga aattcgacac ggagacaaaa
      841 gattgcaacg aagcggcgaa gaaaatgacc atgaatcgcg atgatctggt agaggaggga
      901 gaggaggaaa agtcgaagat tactgaacag aacaatgggt ctaccaaaag tatcaaaaag
      961 atgaagcaca aagctaagaa agaagagaac aatttcagca atgacagcag taaagtcaca
     1021 aaagaactgg agaaaacgga ttacattcac gtgagggcgc gacgagggca ggctacagat
     1081 tcacattcaa ttgcggagag agtacggaga gagaaaatct cagaaaggat gaagttcctc
     1141 caagaccttg tgccaggttg tgacaagatc acaggcaaag caggaatgct ggatgagatc
     1201 atcaactacg tccaatcgtt gcaaagacaa attgagtttc tctcgatgaa actggccatc
     1261 gtgaatccta gaccggattt cgacatggat gacatctttg cgaaagaagt ggcatccact
     1321 cccatgacgg ttgtgccctc accggagatg gtcttgtctg gttacagcca cgaaatggtg
     1381 cattcgggtt attcaagcga gatggtcaat tcgggatacc ttcacgtcaa tcccatgcag
     1441 caggtgaata cttccagtag cggcagcgag actcccggga cctcagagtc cgccacaccc
     1501 gaaagtgata aaaagtattc tattggttta gctatcggca ctaattccgt tggatgggct
     1561 gtcataaccg atgaatacaa agtaccttca aagaaattta aggtgttggg gaacacagac
     1621 cgtcattcga ttaaaaagaa tcttatcggt gccctcctat tcgatagtgg cgaaacggca
     1681 gaggcgactc gcctgaaacg aaccgctcgg agaaggtata cacgtcgcaa gaaccgaata
     1741 tgttacttac aagaaatttt tagcaatgag atggccaaag ttgacgattc tttctttcac
     1801 cgtttggaag agtccttcct tgtcgaagag gacaagaaac atgaacggca ccccatcttt
     1861 ggaaacatag tagatgaggt ggcatatcat gaaaagtacc caacgattta tcacctcaga
     1921 aaaaagctag ttgactcaac tgataaagcg gacctgaggt taatctactt ggctcttgcc
     1981 catatgataa agttccgtgg gcactttctc attgagggtg atctaaatcc ggacaactcg
     2041 gatgtcgaca aactgttcat ccagttagta caaacctata atcagttgtt tgaagagaac
     2101 cctataaatg caagtggcgt ggatgcgaag gctattctta gcgcccgcct ctctaaatcc
     2161 cgacggctag aaaacctgat cgcacaatta cccggagaga agaaaaatgg gttgttcggt
     2221 aaccttatag cgctctcact aggcctgaca ccaaatttta agtcgaactt cgacttagct
     2281 gaagatgcca aattgcagct tagtaaggac acgtacgatg acgatctcga caatctactg
     2341 gcacaaattg gagatcagta tgcggactta tttttggctg ccaaaaacct tagcgatgca
     2401 atcctcctat ctgacatact gagagttaat actgagatta ccaaggcgcc gttatccgct
     2461 tcaatgatca aaaggtacga tgaacatcac caagacttga cacttctcaa ggccctagtc
     2521 cgtcagcaac tgcctgagaa atataaggaa atattctttg atcagtcgaa aaacgggtac
     2581 gcaggttata ttgacggcgg agcgagtcaa gaggaattct acaagtttat caaacccata
     2641 ttagagaaga tggatgggac ggaagagttg cttgtaaaac tcaatcgcga agatctactg
     2701 cgaaagcagc ggactttcga caacggtagc attccacatc aaatccactt aggcgaattg
     2761 catgctatac ttagaaggca ggaggatttt tatccgttcc tcaaagacaa tcgtgaaaag
     2821 attgagaaaa tcctaacctt tcgcatacct tactatgtgg gacccctggc ccgagggaac
     2881 tctcggttcg catggatgac aagaaagtcc gaagaaacga ttactccatg gaattttgag
     2941 gaagttgtcg ataaaggtgc gtcagctcaa tcgttcatcg agaggatgac caactttgac
     3001 aagaatttac cgaacgaaaa agtattgcct aagcacagtt tactttacga gtatttcaca
     3061 gtgtacaatg aactcacgaa agttaagtat gtcactgagg gcatgcgtaa acccgccttt
     3121 ctaagcggag aacagaagaa agcaatagta gatctgttat tcaagaccaa ccgcaaagtg
     3181 acagttaagc aattgaaaga ggactacttt aagaaaattg aatgcttcga ttctgtcgag
     3241 atctccgggg tagaagatcg atttaatgcg tcacttggta cgtatcatga cctcctaaag
     3301 ataattaaag ataaggactt cctggataac gaagagaatg aagatatctt agaagatata
     3361 gtgttgactc ttaccctctt tgaagatcgg gaaatgattg aggaaagact aaaaacatac
     3421 gctcacctgt tcgacgataa ggttatgaaa cagttaaaga ggcgtcgcta tacgggctgg
     3481 ggacgattgt cgcggaaact tatcaacggg ataagagaca agcaaagtgg taaaactatt
     3541 ctcgattttc taaagagcga cggcttcgcc aataggaact ttatgcagct gatccatgat
     3601 gactctttaa ccttcaaaga ggatatacaa aaggcacagg tttccggaca aggggactca
     3661 ttgcacgaac atattgcgaa tcttgctggt tcgccagcca tcaaaaaggg catactccag
     3721 acagtcaaag tagtggatga gctagttaag gtcatgggac gtcacaaacc ggaaaacatt
     3781 gtaatcgaga tggcacgcga aaatcaaacg actcagaagg ggcaaaaaaa cagtcgagag
     3841 cggatgaaga gaatagaaga gggtattaaa gaactgggca gccagatctt aaaggagcat
     3901 cctgtggaaa atacccaatt gcagaacgag aaactttacc tctattacct acaaaatgga
     3961 agggacatgt atgttgatca ggaactggac ataaaccgtt tatctgatta cgacgtcgat
     4021 gccattgtac cccaatcctt tttgaaggac gattcaatcg acaataaagt gcttacacgc
     4081 tcggataaga accgagggaa aagtgacaat gttccaagcg aggaagtcgt aaagaaaatg
     4141 aagaactatt ggcggcagct cctaaatgcg aaactgataa cgcaaagaaa gttcgataac
     4201 ttaactaaag ctgagagggg tggcttgtct gaacttgaca aggccggatt tattaaacgt
     4261 cagctcgtgg aaacccgcca aatcacaaag catgttgcac agatactaga ttcccgaatg
     4321 aatacgaaat acgacgagaa cgataagctg attcgggaag tcaaagtaat cactttaaag
     4381 tcaaaattgg tgtcggactt cagaaaggat tttcaattct ataaagttag ggagataaat
     4441 aactaccacc atgcgcacga cgcttatctt aatgccgtcg tagggaccgc actcattaag
     4501 aaatacccga agctagaaag tgagtttgtg tatggtgatt acaaagttta tgacgtccgt
     4561 aagatgatcg cgaaaagcga acaggagata ggcaaggcta cagccaaata cttcttttat
     4621 tctaacatta tgaatttctt taagacggaa atcactctgg caaacggaga gatacgcaaa
     4681 cgacctttaa ttgaaaccaa tggggagaca ggtgaaatcg tatgggataa gggccgggac
     4741 ttcgcgacgg tgagaaaagt tttgtccatg ccccaagtca acatagtaaa gaaaactgag
     4801 gtgcagaccg gagggttttc aaaggaatcg attcttccaa aaaggaatag tgataagctc
     4861 atcgctcgta aaaaggactg ggacccgaaa aagtacggtg gcttcgatag ccctacagtt
     4921 gcctattctg tcctagtagt ggcaaaagtt gagaagggaa aatccaagaa actgaagtca
     4981 gtcaaagaat tattggggat aacgattatg gagcgctcgt cttttgaaaa gaaccccatc
     5041 gacttccttg aggcgaaagg ttacaaggaa gtaaaaaagg atctcataat taaactacca
     5101 aagtatagtc tgtttgagtt agaaaatggc cgaaaacgga tgttggctag cgccggagag
     5161 cttcaaaagg ggaacgaact cgcactaccg tctaaatacg tgaatttcct gtatttagcg
     5221 tcccattacg agaagttgaa aggttcacct gaagataacg aacagaagca actttttgtt
     5281 gagcagcaca aacattatct cgacgaaatc atagagcaaa tttcggaatt cagtaagaga
     5341 gtcatcctag ctgatgccaa tctggacaaa gtattaagcg catacaacaa gcacagggat
     5401 aaacccatac gtgagcaggc ggaaaatatt atccatttgt ttactcttac caacctcggc
     5461 gctccagccg cattcaagta ttttgacaca acgatagatc gcaaacgata cacttctacc
     5521 aaggaggtgc tagacgcgac actgattcac caatccatca cgggattata tgaaactcgg
     5581 atagatttgt cacagcttgg gggtgactga ccggtcatca tcaccatcac cattgagttt
     5641 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     5701 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     5761 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     5821 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     5881 ctatggcttc tgaggcggaa agaaccagct ggggctcgat accgtcgacc tctagctaga
     5941 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     6001 cacacaacat acgagccgga agcataaagt gtaaagccta gggtgcctaa tgagtgagct
     6061 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     6121 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6181 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6241 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6301 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6361 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6421 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6481 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6541 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6601 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6661 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6721 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6781 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6841 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6901 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6961 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7021 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     7081 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7141 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7201 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7261 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7321 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7381 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7441 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7501 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7561 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7621 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7681 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7741 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7801 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7861 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7921 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7981 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     8041 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     8101 tgccacctga cgtcgacgga tcgggagatc gatctcccga tcccctaggg tcgactctca
     8161 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     8221 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     8281 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca
     8341 gatatacgcg tgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
     8401 ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata
     8461 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
     8521 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
     8581 tgtggaaagg acgaaacacc gggtcttcga gaagacctgt tttagagcta gaaatagcaa
     8641 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
     8701 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgtttttagc gactagttat
     8761 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     8821 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     8881 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     8941 gactatttac ggtaaactgc ccacttggca gtacatcaag tgtatc
//
