LOCUS       sequence-490749-       10163 bp    DNA     circular SYN 27-OCT-2025
DEFINITION  CaMV 35S driving the expression of dCas9-ZAT10(1x).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10163)
  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 10163)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 27, 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: LLP1017
FEATURES             Location/Qualifiers
     source          1..10163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(80..97)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(251..839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(331..350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1010..1801,1802..1870))
                     /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     1633..1652
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1871..1975)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2001..2456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2138..2159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2350..2369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2583..2605
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2597..2614
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2598..2614
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2621..2640
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2621..2639
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2675..3019
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     protein_bind    3020..3039
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     misc_RNA        3060..3135
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    3136..3155
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     terminator      3185..3385
                     /label=gene 7 terminator
                     /note="transcription terminator for octopine-type Ti 
                     plasmid gene 7"
     CDS             3751..3771
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4577..4625
                     /note="synthetic polyadenylation signal"
     misc_feature    4639..4730
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4679..4698
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        4737..5081
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             5097..5117
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(5133..6569,6759..9422)
                     /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          6570..6758
                     /label=IV2 intron
                     /note="modified second intron of the potato ST-LS1 gene 
                     (Vancanneyt et al., 1990)"
     CDS             9435..9455
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9459..9479
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      9696..9948
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(9973..9991)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10012..10028)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10012..10028)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10025..10047)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10036..10052
                     /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(10060..10066,10067..10084,10085..10090))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10105..10126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
       61 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
      121 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
      181 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
      241 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
      301 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
      361 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
      421 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
      481 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
      541 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
      601 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
      661 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
      721 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
      781 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
      841 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
      901 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
      961 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     1021 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     1081 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     1141 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     1201 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     1261 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     1321 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     1381 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     1441 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     1501 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     1561 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     1621 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     1681 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     1741 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     1801 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     1861 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     1921 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     1981 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     2041 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2101 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2161 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2221 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2281 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     2341 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     2401 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
     2461 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     2521 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     2581 ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact cactataggg
     2641 cgaattgggt acaagcttat ctaacgatga gctctgagac ttttcaacaa agggtaatat
     2701 cgggaaacct cctcggattc cattgcccag ctatctgtca cttcatcaaa aggacagtag
     2761 aaaaggaagg tggcacctac aaatgccatc attgcgataa aggaaaggct atcgttcaag
     2821 atgcctctgc cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa
     2881 aagaagacgt tccaaccacg tcttcaaagc aagtggattg atgtgatatc tccactgacg
     2941 taagggatga cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt
     3001 catttcattt ggagaggacg ttcactgccg tataggcagt atagaaccga tcctcccatg
     3061 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
     3121 gcaccgagtc ggtgcgttca ctgccgtata ggcagctgca gactgactaa ctaggatgag
     3181 ctaagctagc tatatcatca atttatgtat tacacataat atcgcactca gtctttcatc
     3241 tacggcaatg taccagctga tataatcagt tattgaaata tttctgaatt taaacttgca
     3301 tcaataaatt tatgtttttg cttggactat aatacctgac ttgttatttt atcaataaat
     3361 atttaaacta tatttctttc aagatgggaa ttaacatcta caaattgcct tttcttatcg
     3421 accatgtacc atatgccaac actcgaatcc ccacccgttg gaccaaaccc ggctcattaa
     3481 gcgtcggttc agatttattt cctttattta aaaaaaagga aagggtaaaa aatagaaaat
     3541 tggaaacagt taaagcccaa aattgtaatt taccgagaat tgtaaattta cctgaaaacc
     3601 ctacgctata gtttcgacta taaataccaa acttaggacc tcacttcaga atcccctcgt
     3661 cgctgcgtct ctctcccgca accttcgatt ttcgtttatt cgcatccatc ggagagagaa
     3721 aacaatcaat aagcgacccc taggatggct ccaaaaaaga aaagaaaggt tgctagcgac
     3781 cattacttag atattagatt gagaccagac ccagagttcc ctccagcaca gttgatgtca
     3841 gtgttattcg gaaaattgca ccaggctttg gttgcacaag gtggagatag aattggtgtg
     3901 agttttcctg atttggacga gtctaggtca agattgggag aaaggttaag aatccatgct
     3961 tcagcagatg accttagggc tttgttagca agaccttggt tagagggtct tagagatcat
     4021 ttgcagttcg gagaaccagc tgttgtgcca caccctacac catatagaca agtttctaga
     4081 gtgcaggcaa aatcaaatcc tgagagactt agaaggagat tgatgaggag acacgactta
     4141 tctgaagagg aagctaggaa gagaatacct gataccgttg ctagagcact tgacttgcca
     4201 tttgtgactt tgagaagtca atccactggt cagcatttta ggttattcat tagacacgga
     4261 ccacttcaag ttactgctga ggagggagga ttcacctgtt atggattgag taagggagga
     4321 ttcgtgcctt ggttctgagg tacccggcca tgctagagtc cgcaaaaatc accagtctct
     4381 ctctacaaat ctatctctct ctatttttct ccagaataat gtgtgagtag ttcccagata
     4441 agggaattag ggttcttata gggtttcgct catgtgttga gcatataaga aacccttagt
     4501 atgtatttgt atttgtaaaa tacttctatc aataaaattt ctaattccta aaaccaaaat
     4561 ccagtgacct actagaaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     4621 gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact
     4681 agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct gaattctgag
     4741 acttttcaac aaagggtaat atcgggaaac ctcctcggat tccattgccc agctatctgt
     4801 cacttcatca aaaggacagt agaaaaggaa ggtggcacct acaaatgcca tcattgcgat
     4861 aaaggaaagg ctatcgttca agatgcctct gccgacagtg gtcccaaaga tggaccccca
     4921 cccacgagga gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat
     4981 tgatgtgata tctccactga cgtaagggat gacgcacaat cccactatcc ttcgcaagac
     5041 ccttcctcta tataaggaag ttcatttcat ttggagagga cggcactagt gatatgccaa
     5101 agaagaagag aaaggtcgga ggaggaggat ctgacaagaa gtactctatc ggacttgcca
     5161 tcggaaccaa ctctgttgga tgggctgtta tcaccgatga gtacaaggtt ccatctaaga
     5221 agttcaaggt tcttggaaac accgatagac actctatcaa gaagaacctt atcggtgctc
     5281 ttcttttcga ttctggagag accgctgagg ctaccagatt gaagagaacc gctagaagaa
     5341 gatacaccag aagaaagaac agaatctgct accttcagga aatcttctct aacgagatgg
     5401 ctaaggttga tgattctttc ttccacagac ttgaggagtc tttccttgtt gaggaggata
     5461 agaagcacga gagacaccca atcttcggaa acatcgttga tgaggttgct taccacgaga
     5521 agtacccaac catctaccac cttagaaaga agttggttga ttctaccgat aaggctgatc
     5581 ttagacttat ctaccttgct cttgctcaca tgatcaagtt cagaggacac ttccttatcg
     5641 agggagacct taacccagat aactctgatg ttgataagtt gttcatccag cttgttcaga
     5701 cctacaacca gcttttcgag gagaacccaa tcaacgcttc tggagttgat gctaaggcta
     5761 tcctttctgc tagactttct aagtctcgta gacttgagaa ccttatcgct cagcttccag
     5821 gagagaagaa gaacggactt ttcggaaacc ttatcgctct ttctcttgga cttaccccaa
     5881 acttcaagtc taacttcgat cttgctgagg atgctaagtt gcagctttct aaggatacct
     5941 acgatgatga tcttgataac cttcttgctc agatcggaga tcagtacgct gatcttttcc
     6001 ttgctgctaa gaacctttct gatgctatcc ttctttctga catccttaga gttaacaccg
     6061 agatcaccaa ggctccactt tctgcttcta tgatcaagag atacgatgag caccaccagg
     6121 atcttaccct tttgaaggct cttgttagac agcagcttcc agagaagtac aaggaaatct
     6181 tcttcgatca gtctaagaac ggatacgctg gatacatcga tggaggagct tctcaggagg
     6241 agttctacaa gttcatcaag ccaatccttg agaagatgga tggaaccgag gagcttcttg
     6301 ttaagttgaa cagagaggat cttcttagaa agcagagaac cttcgataac ggatctatcc
     6361 cacaccagat ccaccttgga gagcttcacg ctatccttcg tagacaggag gatttctacc
     6421 cattcttgaa ggataacaga gagaagatcg agaagatcct taccttcaga atcccatact
     6481 acgttggacc acttgctaga ggaaactctc gtttcgcttg gatgaccaga aagtctgagg
     6541 agaccatcac cccttggaac ttcgaggagg taagtttctg cttctacctt tgatatatat
     6601 ataataatta tcattaatta gtagtaatat aatatttcaa atattttttt caaaataaaa
     6661 gaatgtagta tatagcaatt gcttttctgt agtttataag tgtgtatatt ttaatttata
     6721 acttttctaa tatatgacca aaatttgttg atgtgcaggt tgttgataag ggagcttctg
     6781 ctcagtcttt catcgagaga atgaccaact tcgataagaa ccttccaaac gagaaggttc
     6841 ttccaaagca ctctcttctt tacgagtact tcaccgttta caacgagctt accaaggtta
     6901 agtacgttac cgagggaatg agaaagccag ctttcctttc tggagagcag aagaaggcta
     6961 tcgttgatct tcttttcaag accaacagaa aggttaccgt taagcagttg aaggaggatt
     7021 acttcaagaa gatcgagtgc ttcgattctg ttgaaatctc tggagttgag gatagattca
     7081 acgcttctct tggaacctac cacgatcttt tgaagatcat caaggataag gatttccttg
     7141 ataacgagga gaacgaggac atccttgagg acatcgttct tacccttacc cttttcgagg
     7201 atagagagat gatcgaggag agactcaaga cctacgctca ccttttcgat gataaggtta
     7261 tgaagcagtt gaagagaaga agatacaccg gatggggtag actttctcgt aagttgatca
     7321 acggaatcag agataagcag tctggaaaga ccatccttga tttcttgaag tctgatggat
     7381 tcgctaacag aaacttcatg cagcttatcc acgatgattc tcttaccttc aaggaggaca
     7441 tccagaaggc tcaggtttct ggacagggag attctcttca cgagcacatc gctaaccttg
     7501 ctggatctcc agctatcaag aagggaatcc ttcagaccgt taaggttgtt gatgagcttg
     7561 ttaaggttat gggtagacac aagccagaga acatcgttat cgagatggct agagagaacc
     7621 agaccaccca gaagggacag aagaactctc gtgagagaat gaagagaatc gaggagggaa
     7681 tcaaggagct tggatctcaa atcttgaagg agcacccagt tgagaacacc cagcttcaga
     7741 acgagaagtt gtacctttac taccttcaga acggaagaga tatgtacgtt gatcaggagc
     7801 ttgacatcaa cagactttct gattacgatg ttgatgctat cgttccacag tctttcttga
     7861 aggatgattc tatcgataac aaggttctta cccgttctga taagaacaga ggaaagtctg
     7921 ataacgttcc atctgaggag gttgttaaga agatgaagaa ctactggaga cagcttctta
     7981 acgctaagtt gatcacccag agaaagttcg ataaccttac caaggctgag agaggaggac
     8041 tttctgagct tgataaggct ggattcatca agagacagct tgttgagacc agacagatca
     8101 ccaagcacgt tgctcagatc cttgattctc gtatgaacac caagtacgat gagaacgata
     8161 agttgatcag agaggttaag gttatcacct tgaagtctaa gttggtttct gatttcagaa
     8221 aggatttcca gttctacaag gttagagaga tcaacaacta ccaccacgct cacgatgctt
     8281 accttaacgc tgttgttgga accgctctta tcaagaagta cccaaagttg gagtctgagt
     8341 tcgtttacgg agattacaag gtttacgatg ttagaaagat gatcgctaag tctgagcagg
     8401 agatcggaaa ggctaccgct aagtacttct tctactctaa catcatgaac ttcttcaaga
     8461 ccgagatcac ccttgctaac ggagagatca gaaagagacc acttatcgag accaacggag
     8521 agaccggaga gatcgtttgg gataagggaa gagatttcgc taccgttaga aaggttcttt
     8581 ctatgccaca ggttaacatc gttaagaaaa ccgaggttca gaccggagga ttctctaagg
     8641 agtctatcct tccaaagaga aactctgata agttgatcgc tagaaagaag gattgggacc
     8701 caaagaagta cggaggattc gattctccaa ccgttgctta ctctgttctt gttgttgcta
     8761 aggttgagaa gggaaagtct aagaagttga agtctgttaa ggagcttctt ggaatcacca
     8821 tcatggagcg ttcttctttc gagaagaacc caatcgattt ccttgaggct aagggataca
     8881 aggaggttaa gaaggatctt atcatcaagt tgccaaagta ctctcttttc gagcttgaga
     8941 acggaagaaa gagaatgctt gcttctgctg gagagcttca gaagggaaac gagcttgctc
     9001 ttccatctaa gtacgttaac ttcctttacc ttgcttctca ctacgagaag ttgaagggat
     9061 ctccagagga taacgagcag aagcagcttt tcgttgagca gcacaagcac taccttgatg
     9121 agatcatcga gcaaatctct gagttctcta agagagttat ccttgctgat gctaaccttg
     9181 ataaggttct ttctgcttac aacaagcaca gagataagcc aatcagagag caggctgaga
     9241 acatcatcca ccttttcacc cttaccaacc ttggtgctcc agctgctttc aagtacttcg
     9301 ataccaccat cgatagaaaa agatacacct ctaccaagga ggttcttgat gctaccctta
     9361 tccaccagtc tatcaccgga ctttacgaga ccagaatcga tctttctcag cttggaggag
     9421 atgaaggagc tgatcctaag aagaaaagaa aggttgatcc aaagaaaaag aggaaagttg
     9481 gtggaggtgg atcaggaggt ggaggatctg gtgggggagg atccgaggga gcaggatcaa
     9541 cttcacatgt gtcttcatct cacagaggtt ttgaccttaa cattccacct atccctgagt
     9601 tttcaatggt gaatggtgat gacgaagtga tgtcacctat gccagctaaa aagccaagat
     9661 tcgatttccc tgtgaagttg caattgtgag tcgacgatcg ttcaaacatt tggcaataaa
     9721 gtttcttaag attgaatcct gttgccggtc ttgcgacgat tatcatataa tttctgttga
     9781 attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg agatgggttt
     9841 ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa atatagcgcg
     9901 caaactagga taaattatcg cgcgcggtgt catctatgtt actagatcag tcactcgagc
     9961 cagcttttgt tccctttagt gagggttaat ttcgagcttg gcgtaatcat ggtcatagct
    10021 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
    10081 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
    10141 actgcccgct ttccagtcgg gaa
//
