LOCUS       sequence-490744-        9944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CaMV 35S driving the expression of dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9944)
  AUTHORS   Khan MA, Herring G, Zhu JY, Oliva M, Fourie E, Johnston B, Zhang Z, 
            Potter J, Pineda L, Pflueger J, Swain T, Pflueger C, Lloyd JPB, 
            Secco D, Small I, Kidd BN, Lister R
  TITLE     CRISPRi-based circuits to control gene expression in plants.
  JOURNAL   Nat Biotechnol. 2025 Mar;43(3):416-430. doi: 
            10.1038/s41587-024-02236-w. Epub 2024 May 20.
  PUBMED    38769424
REFERENCE   2  (bases 1 to 9944)
  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: 
            "10.1038/s41587-024-02236-w"; journalName: "Nat Biotechnol"; date: 
            "2025-03"; volume: "43"; issue: "3"; pages: "416-430"
COMMENT     Sequence Label: LLP1023
FEATURES             Location/Qualifiers
     source          1..9944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(99..687)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(179..198)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(858..1649,1650..1718))
                     /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     1481..1500
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1719..1823)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1849..2304)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1986..2007)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2198..2217
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2431..2453
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2445..2462
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2446..2462
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2469..2488
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2469..2487
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2523..2867
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     protein_bind    2868..2887
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        2908..2983
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    2984..3003
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     terminator      3033..3233
                     /label=gene 7 terminator
                     /note="transcription terminator for octopine-type Ti 
                     plasmid gene 7"
     promoter        3284..3628
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             3641..3661
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4467..4515
                     /note="synthetic polyadenylation signal"
     misc_feature    4529..4620
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4569..4588
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        4627..4971
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             4987..5007
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(5020..6456,6646..9309)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system"
                     /label=pcoCas9
                     /note="plant codon-optimized Cas9 gene containing the 
                     potato IV2 intron"
                     /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"
     intron          6457..6645
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             9322..9342
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(9602..9620)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(9641..9657)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9641..9657)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9654..9676)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9665..9681
                     /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(9689..9695,9696..9713,9714..9719))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9734..9755
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9872..9889)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
       61 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
      121 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
      181 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
      241 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
      301 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
      361 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
      421 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
      481 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
      541 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
      601 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
      661 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
      721 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
      781 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
      841 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
      901 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
      961 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     1021 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     1081 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     1141 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     1201 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     1261 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     1321 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     1381 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     1441 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     1501 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     1561 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     1621 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     1681 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     1741 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     1801 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     1861 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     1921 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     1981 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     2041 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     2101 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     2161 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     2221 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     2281 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     2341 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     2401 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     2461 gtgaattgta atacgactca ctatagggcg aattgggtac aagcttatct aacgatgagc
     2521 tctgagactt ttcaacaaag ggtaatatcg ggaaacctcc tcggattcca ttgcccagct
     2581 atctgtcact tcatcaaaag gacagtagaa aaggaaggtg gcacctacaa atgccatcat
     2641 tgcgataaag gaaaggctat cgttcaagat gcctctgccg acagtggtcc caaagatgga
     2701 cccccaccca cgaggagcat cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa
     2761 gtggattgat gtgatatctc cactgacgta agggatgacg cacaatccca ctatccttcg
     2821 caagaccctt cctctatata aggaagttca tttcatttgg agaggacgtt cactgccgta
     2881 taggcagtat agaaccgatc ctcccatgtt ttagagctag aaatagcaag ttaaaataag
     2941 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcgttcact gccgtatagg
     3001 cagctgcaga ctgactaact aggatgagct aagctagcta tatcatcaat ttatgtatta
     3061 cacataatat cgcactcagt ctttcatcta cggcaatgta ccagctgata taatcagtta
     3121 ttgaaatatt tctgaattta aacttgcatc aataaattta tgtttttgct tggactataa
     3181 tacctgactt gttattttat caataaatat ttaaactata tttctttcaa gatgggaatt
     3241 aacatctaca aattgccttt tcttatcgac catgtaccat atgtgagact tttcaacaaa
     3301 gggtaatatc gggaaacctc ctcggattcc attgcccagc tatctgtcac ttcatcaaaa
     3361 ggacagtaga aaaggaaggt ggcacctaca aatgccatca ttgcgataaa ggaaaggcta
     3421 tcgttcaaga tgcctctgcc gacagtggtc ccaaagatgg acccccaccc acgaggagca
     3481 tcgtggaaaa agaagacgtt ccaaccacgt cttcaaagca agtggattga tgtgatatct
     3541 ccactgacgt aagggatgac gcacaatccc actatccttc gcaagaccct tcctctatat
     3601 aaggaagttc atttcatttg gagaggaccc taggatggct ccaaaaaaga aaagaaaggt
     3661 tgctagcgac cattacttag atattagatt gagaccagac ccagagttcc ctccagcaca
     3721 gttgatgtca gtgttattcg gaaaattgca ccaggctttg gttgcacaag gtggagatag
     3781 aattggtgtg agttttcctg atttggacga gtctaggtca agattgggag aaaggttaag
     3841 aatccatgct tcagcagatg accttagggc tttgttagca agaccttggt tagagggtct
     3901 tagagatcat ttgcagttcg gagaaccagc tgttgtgcca caccctacac catatagaca
     3961 agtttctaga gtgcaggcaa aatcaaatcc tgagagactt agaaggagat tgatgaggag
     4021 acacgactta tctgaagagg aagctaggaa gagaatacct gataccgttg ctagagcact
     4081 tgacttgcca tttgtgactt tgagaagtca atccactggt cagcatttta ggttattcat
     4141 tagacacgga ccacttcaag ttactgctga ggagggagga ttcacctgtt atggattgag
     4201 taagggagga ttcgtgcctt ggttctgagg tacccggcca tgctagagtc cgcaaaaatc
     4261 accagtctct ctctacaaat ctatctctct ctatttttct ccagaataat gtgtgagtag
     4321 ttcccagata agggaattag ggttcttata gggtttcgct catgtgttga gcatataaga
     4381 aacccttagt atgtatttgt atttgtaaaa tacttctatc aataaaattt ctaattccta
     4441 aaaccaaaat ccagtgacct actagaaata aaatatcttt attttcatta catctgtgtg
     4501 ttggtttttt gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac
     4561 aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct
     4621 gaattctgag acttttcaac aaagggtaat atcgggaaac ctcctcggat tccattgccc
     4681 agctatctgt cacttcatca aaaggacagt agaaaaggaa ggtggcacct acaaatgcca
     4741 tcattgcgat aaaggaaagg ctatcgttca agatgcctct gccgacagtg gtcccaaaga
     4801 tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa
     4861 gcaagtggat tgatgtgata tctccactga cgtaagggat gacgcacaat cccactatcc
     4921 ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga cggcactagt
     4981 gatatgccaa agaagaagcg gaaggtcggg ggtggttctg acaagaagta ctctatcgga
     5041 cttgccatcg gaaccaactc tgttggatgg gctgttatca ccgatgagta caaggttcca
     5101 tctaagaagt tcaaggttct tggaaacacc gatagacact ctatcaagaa gaaccttatc
     5161 ggtgctcttc ttttcgattc tggagagacc gctgaggcta ccagattgaa gagaaccgct
     5221 agaagaagat acaccagaag aaagaacaga atctgctacc ttcaggaaat cttctctaac
     5281 gagatggcta aggttgatga ttctttcttc cacagacttg aggagtcttt ccttgttgag
     5341 gaggataaga agcacgagag acacccaatc ttcggaaaca tcgttgatga ggttgcttac
     5401 cacgagaagt acccaaccat ctaccacctt agaaagaagt tggttgattc taccgataag
     5461 gctgatctta gacttatcta ccttgctctt gctcacatga tcaagttcag aggacacttc
     5521 cttatcgagg gagaccttaa cccagataac tctgatgttg ataagttgtt catccagctt
     5581 gttcagacct acaaccagct tttcgaggag aacccaatca acgcttctgg agttgatgct
     5641 aaggctatcc tttctgctag actttctaag tctcgtagac ttgagaacct tatcgctcag
     5701 cttccaggag agaagaagaa cggacttttc ggaaacctta tcgctctttc tcttggactt
     5761 accccaaact tcaagtctaa cttcgatctt gctgaggatg ctaagttgca gctttctaag
     5821 gatacctacg atgatgatct tgataacctt cttgctcaga tcggagatca gtacgctgat
     5881 cttttccttg ctgctaagaa cctttctgat gctatccttc tttctgacat ccttagagtt
     5941 aacaccgaga tcaccaaggc tccactttct gcttctatga tcaagagata cgatgagcac
     6001 caccaggatc ttaccctttt gaaggctctt gttagacagc agcttccaga gaagtacaag
     6061 gaaatcttct tcgatcagtc taagaacgga tacgctggat acatcgatgg aggagcttct
     6121 caggaggagt tctacaagtt catcaagcca atccttgaga agatggatgg aaccgaggag
     6181 cttcttgtta agttgaacag agaggatctt cttagaaagc agagaacctt cgataacgga
     6241 tctatcccac accagatcca ccttggagag cttcacgcta tccttcgtag acaggaggat
     6301 ttctacccat tcttgaagga taacagagag aagatcgaga agatccttac cttcagaatc
     6361 ccatactacg ttggaccact tgctagagga aactctcgtt tcgcttggat gaccagaaag
     6421 tctgaggaga ccatcacccc ttggaacttc gaggaggtaa gtttctgctt ctacctttga
     6481 tatatatata ataattatca ttaattagta gtaatataat atttcaaata tttttttcaa
     6541 aataaaagaa tgtagtatat agcaattgct tttctgtagt ttataagtgt gtatatttta
     6601 atttataact tttctaatat atgaccaaaa tttgttgatg tgcaggttgt tgataaggga
     6661 gcttctgctc agtctttcat cgagagaatg accaacttcg ataagaacct tccaaacgag
     6721 aaggttcttc caaagcactc tcttctttac gagtacttca ccgtttacaa cgagcttacc
     6781 aaggttaagt acgttaccga gggaatgaga aagccagctt tcctttctgg agagcagaag
     6841 aaggctatcg ttgatcttct tttcaagacc aacagaaagg ttaccgttaa gcagttgaag
     6901 gaggattact tcaagaagat cgagtgcttc gattctgttg aaatctctgg agttgaggat
     6961 agattcaacg cttctcttgg aacctaccac gatcttttga agatcatcaa ggataaggat
     7021 ttccttgata acgaggagaa cgaggacatc cttgaggaca tcgttcttac ccttaccctt
     7081 ttcgaggata gagagatgat cgaggagaga ctcaagacct acgctcacct tttcgatgat
     7141 aaggttatga agcagttgaa gagaagaaga tacaccggat ggggtagact ttctcgtaag
     7201 ttgatcaacg gaatcagaga taagcagtct ggaaagacca tccttgattt cttgaagtct
     7261 gatggattcg ctaacagaaa cttcatgcag cttatccacg atgattctct taccttcaag
     7321 gaggacatcc agaaggctca ggtttctgga cagggagatt ctcttcacga gcacatcgct
     7381 aaccttgctg gatctccagc tatcaagaag ggaatccttc agaccgttaa ggttgttgat
     7441 gagcttgtta aggttatggg tagacacaag ccagagaaca tcgttatcga gatggctaga
     7501 gagaaccaga ccacccagaa gggacagaag aactctcgtg agagaatgaa gagaatcgag
     7561 gagggaatca aggagcttgg atctcaaatc ttgaaggagc acccagttga gaacacccag
     7621 cttcagaacg agaagttgta cctttactac cttcagaacg gaagagatat gtacgttgat
     7681 caggagcttg acatcaacag actttctgat tacgatgttg atgctatcgt tccacagtct
     7741 ttcttgaagg atgattctat cgataacaag gttcttaccc gttctgataa gaacagagga
     7801 aagtctgata acgttccatc tgaggaggtt gttaagaaga tgaagaacta ctggagacag
     7861 cttcttaacg ctaagttgat cacccagaga aagttcgata accttaccaa ggctgagaga
     7921 ggaggacttt ctgagcttga taaggctgga ttcatcaaga gacagcttgt tgagaccaga
     7981 cagatcacca agcacgttgc tcagatcctt gattctcgta tgaacaccaa gtacgatgag
     8041 aacgataagt tgatcagaga ggttaaggtt atcaccttga agtctaagtt ggtttctgat
     8101 ttcagaaagg atttccagtt ctacaaggtt agagagatca acaactacca ccacgctcac
     8161 gatgcttacc ttaacgctgt tgttggaacc gctcttatca agaagtaccc aaagttggag
     8221 tctgagttcg tttacggaga ttacaaggtt tacgatgtta gaaagatgat cgctaagtct
     8281 gagcaggaga tcggaaaggc taccgctaag tacttcttct actctaacat catgaacttc
     8341 ttcaagaccg agatcaccct tgctaacgga gagatcagaa agagaccact tatcgagacc
     8401 aacggagaga ccggagagat cgtttgggat aagggaagag atttcgctac cgttagaaag
     8461 gttctttcta tgccacaggt taacatcgtt aagaaaaccg aggttcagac cggaggattc
     8521 tctaaggagt ctatccttcc aaagagaaac tctgataagt tgatcgctag aaagaaggat
     8581 tgggacccaa agaagtacgg aggattcgat tctccaaccg ttgcttactc tgttcttgtt
     8641 gttgctaagg ttgagaaggg aaagtctaag aagttgaagt ctgttaagga gcttcttgga
     8701 atcaccatca tggagcgttc ttctttcgag aagaacccaa tcgatttcct tgaggctaag
     8761 ggatacaagg aggttaagaa ggatcttatc atcaagttgc caaagtactc tcttttcgag
     8821 cttgagaacg gaagaaagag aatgcttgct tctgctggag agcttcagaa gggaaacgag
     8881 cttgctcttc catctaagta cgttaacttc ctttaccttg cttctcacta cgagaagttg
     8941 aagggatctc cagaggataa cgagcagaag cagcttttcg ttgagcagca caagcactac
     9001 cttgatgaga tcatcgagca aatctctgag ttctctaaga gagttatcct tgctgatgct
     9061 aaccttgata aggttctttc tgcttacaac aagcacagag ataagccaat cagagagcag
     9121 gctgagaaca tcatccacct tttcaccctt accaaccttg gtgctccagc tgctttcaag
     9181 tacttcgata ccaccatcga tagaaaaaga tacacctcta ccaaggaggt tcttgatgct
     9241 acccttatcc accagtctat caccggactt tacgagacca gaatcgatct ttctcagctt
     9301 ggaggagatg gtgggggcag tcctaaaaag aaaaggaagg tgtaagtcga ccggccatgc
     9361 tagagtccgc aaaaatcacc agtctctctc tacaaatcta tctctctcta tttttctcca
     9421 gaataatgtg tgagtagttc ccagataagg gaattagggt tcttataggg tttcgctcat
     9481 gtgttgagca tataagaaac ccttagtatg tatttgtatt tgtaaaatac ttctatcaat
     9541 aaaatttcta attcctaaaa ccaaaatcca gtgacctagt cactcgagcc agcttttgtt
     9601 ccctttagtg agggttaatt tcgagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     9661 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     9721 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     9781 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     9841 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     9901 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aata
//
