LOCUS       sequence-266440-       13239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pGreen3 CRISPR/Cas9 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13239)
  AUTHORS   Zhang Q, Zhang Y, Lu MH, Chai YP, Jiang YY, Zhou Y, Wang XC, Chen QJ
  TITLE     A Novel Ternary Vector System United with Morphogenic Genes Enhances
            CRISPR/Cas Delivery in Maize.
  JOURNAL   Plant Physiol. 2019 Sep 26. pii: pp.19.00767. doi: 
            10.1104/pp.19.00767.
  PUBMED    31558579
REFERENCE   2  (bases 1 to 13239)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant 
            Physiol. 2019 Sep 26. pii: pp.19.00767. doi: 10.1104/pp.19.00767."
COMMENT     Sequence Label: pG3H-U6SC
FEATURES             Location/Qualifiers
     source          1..13239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        473..577
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(578..646,647..1438)
                     /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     complement(796..815)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_RNA        1587..1662
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             2638..2703
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2710..2730
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2755..6855
                     /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             2755..6855
                     /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             6768..6776
                     /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             6856..6903
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(7560..7576)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7560..7576)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7573..7595)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7584..7600
                     /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(7608..7614,7615..7632,7633..7638))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7653..7674
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7865..8543
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     8480..8500
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             8610..9635
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     polyA_signal    9676..9850
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    9928..9952
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(10330..10349)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             10341..11150
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(10408..10427)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      11229..11315
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      11407..11434
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      11513..12101
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12002..12021
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     rep_origin      12216..12826
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      12303..12834
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    13013..13037
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     join(13225..13239,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(13239,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgcc aagcttcgac ttgccttccg cacaatacat catttcttct
       61 tagctttttt tcttcttctt cgttcataca gttttttttt gtttatcagc ttacattttc
      121 ttgaaccgta gctttcgttt tcttcttttt aactttccat tcggagtttt tgtatcttgt
      181 ttcatagttt gtcccaggat tagaatgatt aggcatcgaa ccttcaagaa tttgattgaa
      241 taaaacatct tcattcttaa gatatgaaga taatcttcaa aaggcccctg ggaatctgaa
      301 agaagagaag caggcccatt tatatgggaa agaacaatag tatttcttat ataggcccat
      361 ttaagttgaa aacaatcttc aaaagtccca catcgcttag ataagaaaac gaagctgagt
      421 ttatatacag ctagagtcga agtagtgatt gtgagacctt cggggaaatg tgcgcggaac
      481 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatgg gacaataacc
      541 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
      601 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
      661 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
      721 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
      781 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
      841 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
      901 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
      961 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     1021 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     1081 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     1141 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     1201 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     1261 tattgctgat aaatctggag ccggtgagcg tggctctcgc ggtatcattg cagcactggg
     1321 gccagatggt aagccctccc gtatcgtagt tatctacacc acggggagtc aggcaactat
     1381 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1441 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     1501 aaggatctag gtgaagatcc tttttgataa tcccatgacc aaaatccctt aacgtgagtt
     1561 ttcgttccac tgagcgtcag gtctcagttt tagagctaga aatagcaagt taaaataagg
     1621 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttt ggatagaatt
     1681 tcccagcttt tttgcgtgtt tcagctctca tgatccttgc atgcactagt atggagtcaa
     1741 agattcaaat agaggaccta acagaactcg ccgtaaagac tggcgaacag ttcatacaga
     1801 gtctcttacg actcaatgac aagaagaaaa tcttcgtcaa catggtggag cacgacacac
     1861 ttgtctactc caaaaatatc aaagatacag tctcagaaga ccaaagggca attgagactt
     1921 ttcaacaaag ggtaatatcc ggaaacctcc tcggattcca ttgcccagct atctgtcact
     1981 ttattgtgaa gatagtggaa aaggaaggtg gctcctacaa atgccatcat tgcgataaag
     2041 gaaaggccat cgttgaagat gcctctgccg acagtggtcc caaagatgga cccccaccca
     2101 cgaggagcat cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat
     2161 gtgattggca gacatactgt cccacaaatg aagatggaat ctgtaaaaga aaacgcgtga
     2221 aataatgcgt ctgacaaagg ttaggtcggc tgcctttaat caataccaaa gtggtcccta
     2281 ccacgatgga aaaactgtgc agtcggtttg gctttttctg acgaacaaat aagattcgtg
     2341 gccgacaggt gggggtccac catgtgaagg catcttcaga ctccaataat ggagcaatga
     2401 cgtaagggct tacgaaataa gtaagggtag tttgggaaat gtccactcac ccgtcagtct
     2461 ataaatactt agcccctccc tcattgttaa gggagcaaaa tctcagagag atagtcctag
     2521 agagagaaag agagcaagta gcctagaagt agtcaaggcg gcgaagtatt caggcacgtg
     2581 gccaggaaga agaaaagcca agacgacgaa aacaggtaag agctaagcat ctagatggat
     2641 tacaaggacc acgacgggga ttacaaggac cacgacattg attacaagga tgatgatgac
     2701 aagatggctc cgaagaagaa gaggaaggtt ggcatccacg gggtgccagc tgctgacaag
     2761 aagtactcga tcggcctcga tattgggact aactctgttg gctgggccgt gatcaccgac
     2821 gagtacaagg tgccctcaaa gaagttcaag gtcctgggca acaccgatcg gcattccatc
     2881 aagaagaatc tcattggcgc tctcctgttc gacagcggcg agacggctga ggctacgcgg
     2941 ctcaagcgca ccgcccgcag gcggtacacg cgcaggaaga atcgcatctg ctacctgcag
     3001 gagattttct ccaacgagat ggcgaaggtt gacgattctt tcttccacag gctggaggag
     3061 tcattcctcg tggaggagga taagaagcac gagcggcatc caatcttcgg caacattgtc
     3121 gacgaggttg cctaccacga gaagtaccct acgatctacc atctgcggaa gaagctcgtg
     3181 gactccacag ataaggcgga cctccgcctg atctacctcg ctctggccca catgattaag
     3241 ttcaggggcc atttcctgat cgagggggat ctcaacccgg acaatagcga tgttgacaag
     3301 ctgttcatcc agctcgtgca gacgtacaac cagctcttcg aggagaaccc cattaatgcg
     3361 tcaggcgtcg acgcgaaggc tatcctgtcc gctaggctct cgaagtctcg gcgcctcgag
     3421 aacctgatcg cccagctgcc gggcgagaag aagaacggcc tgttcgggaa tctcattgcg
     3481 ctcagcctgg ggctcacgcc caacttcaag tcgaatttcg atctcgctga ggacgccaag
     3541 ctgcagctct ccaaggacac atacgacgat gacctggata acctcctggc ccagatcggc
     3601 gatcagtacg cggacctgtt cctcgctgcc aagaatctgt cggacgccat cctcctgtct
     3661 gatattctca gggtgaacac cgagattacg aaggctccgc tctcagcctc catgatcaag
     3721 cgctacgacg agcaccatca ggatctgacc ctcctgaagg cgctggtcag gcagcagctc
     3781 cccgagaagt acaaggagat cttcttcgat cagtcgaaga acggctacgc tgggtacatt
     3841 gacggcgggg cctctcagga ggagttctac aagttcatca agccgattct ggagaagatg
     3901 gacggcacgg aggagctgct ggtgaagctc aatcgcgagg acctcctgag gaagcagcgg
     3961 acattcgata acggcagcat cccacaccag attcatctcg gggagctgca cgctatcctg
     4021 aggaggcagg aggacttcta ccctttcctc aaggataacc gcgagaagat cgagaagatt
     4081 ctgactttca ggatcccgta ctacgtcggc ccactcgcta ggggcaactc ccgcttcgct
     4141 tggatgaccc gcaagtcaga ggagacgatc acgccgtgga acttcgagga ggtggtcgac
     4201 aagggcgcta gcgctcagtc gttcatcgag aggatgacga atttcgacaa gaacctgcca
     4261 aatgagaagg tgctccctaa gcactcgctc ctgtacgagt acttcacagt ctacaacgag
     4321 ctgactaagg tgaagtatgt gaccgagggc atgaggaagc cggctttcct gtctggggag
     4381 cagaagaagg ccatcgtgga cctcctgttc aagaccaacc ggaaggtcac ggttaagcag
     4441 ctcaaggagg actacttcaa gaagattgag tgcttcgatt cggtcgagat ctctggcgtt
     4501 gaggaccgct tcaacgcctc cctggggacc taccacgatc tcctgaagat cattaaggat
     4561 aaggacttcc tggacaacga ggagaatgag gatatcctcg aggacattgt gctgacactc
     4621 actctgttcg aggaccggga gatgatcgag gagcgcctga agacttacgc ccatctcttc
     4681 gatgacaagg tcatgaagca gctcaagagg aggaggtaca ccggctgggg gaggctgagc
     4741 aggaagctca tcaacggcat tcgggacaag cagtccggga agacgatcct cgacttcctg
     4801 aagagcgatg gcttcgcgaa ccgcaatttc atgcagctga ttcacgatga cagcctcaca
     4861 ttcaaggagg atatccagaa ggctcaggtg agcggccagg gggactcgct gcacgagcat
     4921 atcgcgaacc tcgctggctc gccagctatc aagaagggga ttctgcagac cgtgaaggtt
     4981 gtggacgagc tggtgaaggt catgggcagg cacaagcctg agaacatcgt cattgagatg
     5041 gcccgggaga atcagaccac gcagaagggc cagaagaact cacgcgagag gatgaagagg
     5101 atcgaggagg gcattaagga gctggggtcc cagatcctca aggagcaccc ggtggagaac
     5161 acgcagctgc agaatgagaa gctctacctg tactacctcc agaatggccg cgatatgtat
     5221 gtggaccagg agctggatat taacaggctc agcgattacg acgtcgatca tatcgttcca
     5281 cagtcattcc tgaaggatga ctccattgac aacaaggtcc tcaccaggtc ggacaagaac
     5341 cggggcaagt ctgataatgt tccttcagag gaggtcgtta agaagatgaa gaactactgg
     5401 cgccagctcc tgaatgccaa gctgatcacg cagcggaagt tcgataacct cacaaaggct
     5461 gagaggggcg ggctctctga gctggacaag gcgggcttca tcaagaggca gctggtcgag
     5521 acacggcaga tcactaagca cgttgcgcag attctcgact cacggatgaa cactaagtac
     5581 gatgagaatg acaagctgat ccgcgaggtg aaggtcatca ccctgaagtc aaagctcgtc
     5641 tccgacttca ggaaggattt ccagttctac aaggttcggg agatcaacaa ttaccaccat
     5701 gcccatgacg cgtacctgaa cgcggtggtc ggcacagctc tgatcaagaa gtacccaaag
     5761 ctcgagagcg agttcgtgta cggggactac aaggtttacg atgtgaggaa gatgatcgcc
     5821 aagtcggagc aggagattgg caaggctacc gccaagtact tcttctactc taacattatg
     5881 aatttcttca agacagagat cactctggcc aatggcgaga tccggaagcg ccccctcatc
     5941 gagacgaacg gcgagacggg ggagatcgtg tgggacaagg gcagggattt cgcgaccgtc
     6001 aggaaggttc tctccatgcc acaagtgaat atcgtcaaga agacagaggt ccagactggc
     6061 gggttctcta aggagtcaat tctgcctaag cggaacagcg acaagctcat cgcccgcaag
     6121 aaggactggg atccgaagaa gtacggcggg ttcgacagcc ccactgtggc ctactcggtc
     6181 ctggttgtgg cgaaggttga gaagggcaag tccaagaagc tcaagagcgt gaaggagctg
     6241 ctggggatca cgattatgga gcgctccagc ttcgagaaga acccgatcga tttcctggag
     6301 gcgaagggct acaaggaggt gaagaaggac ctgatcatta agctccccaa gtactcactc
     6361 ttcgagctgg agaacggcag gaagcggatg ctggcttccg ctggcgagct gcagaagggg
     6421 aacgagctgg ctctgccgtc caagtatgtg aacttcctct acctggcctc ccactacgag
     6481 aagctcaagg gcagccccga ggacaacgag cagaagcagc tgttcgtcga gcagcacaag
     6541 cattacctcg acgagatcat tgagcagatt tccgagttct ccaagcgcgt gatcctggcc
     6601 gacgcgaatc tggataaggt cctctccgcg tacaacaagc accgcgacaa gccaatcagg
     6661 gagcaggctg agaatatcat tcatctcttc accctgacga acctcggcgc ccctgctgct
     6721 ttcaagtact tcgacacaac tatcgatcgc aagaggtaca caagcactaa ggaggtcctg
     6781 gacgcgaccc tcatccacca gtcgattacc ggcctctacg agacgcgcat cgacctgtct
     6841 cagctcgggg gcgacaagcg gccagcggcg acgaagaagg cggggcaggc gaagaagaag
     6901 aagtgagctc agagctttcg ttcgtatcat cggtttcgac aacgttcgtc aagttcaatg
     6961 catcagtttc attgcgcaca caccagaatc ctactgagtt tgagtattat ggcattggga
     7021 aaactgtttt tcttgtacca tttgttgtgc ttgtaattta ctgtgttttt tattcggttt
     7081 tcgctatcga actgtgaaat ggaaatggat ggagaagagt taatgaatga tatggtcctt
     7141 ttgttcattc tcaaattaat attatttgtt ttttctctta tttgttgtgt gttgaatttg
     7201 aaattataag agatatgcaa acattttgtt ttgagtaaaa atgtgtcaaa tcgtggcctc
     7261 taatgaccga agttaatatg aggagtaaaa cacttgtagt tgtaccatta tgcttattca
     7321 ctaggcaaca aatatatttt cagacctaga aaagctgcaa atgttactga atacaagtat
     7381 gtcctcttgt gttttagaca tttatgaact ttcctttatg taattttcca gaatccttgt
     7441 cagattctaa tcattgcttt ataattatag ttatactcat ggatttgtag ttgagtatga
     7501 aaatattttt taatgcattt tatgacttgc caattgattg acaacgaatt cgtaatcatg
     7561 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     7621 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     7681 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     7741 ggccaacgcg cggggagagg cggtttgcgt attggctaga gcagcttgcc aacatggtgg
     7801 agcacgacac tctcgtctac tccaagaata tcaaagatac agtctcagaa gaccaaaggg
     7861 ctattgagac ttttcaacaa agggtaatat cgggaaacct cctcggattc cattgcccag
     7921 ctatctgtca cttcatcaaa aggacagtag aaaaggaagg tggcacctac aaatgccatc
     7981 attgcgataa aggaaaggct atcgttcaag atgcctctgc cgacagtggt cccaaagatg
     8041 gacccccacc cacgaggagc atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc
     8101 aagtggattg atgtgataac atggtggagc acgacactct cgtctactcc aagaatatca
     8161 aagatacagt ctcagaagac caaagggcta ttgagacttt tcaacaaagg gtaatatcgg
     8221 gaaacctcct cggattccat tgcccagcta tctgtcactt catcaaaagg acagtagaaa
     8281 aggaaggtgg cacctacaaa tgccatcatt gcgataaagg aaaggctatc gttcaagatg
     8341 cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc gtggaaaaag
     8401 aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgatatctcc actgacgtaa
     8461 gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa ggaagttcat
     8521 ttcatttgga gaggacacgc tgaaatcacc agtctctctc tacaaatcta tctctctcga
     8581 gctttcgcag atccgggggg caatgagata tgaaaaagcc tgaactcacc gcgacgtctg
     8641 tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg
     8701 gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa
     8761 atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg
     8821 cgctcccgat tccggaagtg cttgacattg gggagtttag cgagagcctg acctattgca
     8881 tctcccgccg ttcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg
     8941 ttctacaacc ggtcgcggag gctatggatg cgatcgctgc ggccgatctt agccagacga
     9001 gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca
     9061 tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca
     9121 gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag
     9181 tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca
     9241 taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca
     9301 acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc
     9361 ggaggcatcc ggagcttgca ggatcgccac gactccgggc gtatatgctc cgcattggtc
     9421 ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg
     9481 gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc
     9541 gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc
     9601 gacgccccag cactcgtccg agggcaaaga aatagagtag atgccgaccg ggatctgtcg
     9661 atcgacaagc tcgagtttct ccataataat gtgtgagtag ttcccagata agggaattag
     9721 ggttcctata gggtttcgct catgtgttga gcatataaga aacccttagt atgtatttgt
     9781 atttgtaaaa tacttctatc aataaaattt ctaattccta aaaccaaaat ccagtactaa
     9841 aatccagatc ccccgaatta attcggcgtt aattcagtac attaaaaacg tccgcaatgt
     9901 gttattaagt tgtctaagcg tcaatttgtt tacaccacaa tatatcctgc caccagccag
     9961 ccaacagctc cccgaccggc agctcggcac aaaatcacca ctcgatacag gcagcccatc
    10021 agtccgggac ggcgtcagcg ggagagccgt tgtaaggcgg cagactttgc tcatgttacc
    10081 gatgctattc ggaagaacgg caactaagct gccgggtttg aaacacggat gatctcgcgg
    10141 agggtagcat gttgattgta acgatgacag agcgttgctg cctgtgatca ccgcggctgg
    10201 caaacagcta ttatgggtat tatgggtcta gtctctggcc cgtgtctcaa aatctctgat
    10261 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
    10321 acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa
    10381 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
    10441 caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac
    10501 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
    10561 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt
    10621 tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt
    10681 caggtgaaaa tattgttgat gcgctggcag tgtccctgcg ccggttgcat tcgattcctg
    10741 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
    10801 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
    10861 aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc
    10921 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
    10981 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
    11041 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
    11101 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg
    11161 ttaattggtt gtaacattat tcagattggc gtctaactac taagcgagag tagggaactg
    11221 ccaggcatca aataaaacga aaggctcagt cggaagactg ggcctttcgt tttatctgtt
    11281 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg
    11341 tgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa
    11401 ctaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact cttcctgtta
    11461 gggccccgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
    11521 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
    11581 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
    11641 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
    11701 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
    11761 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
    11821 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
    11881 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
    11941 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
    12001 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
    12061 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
    12121 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
    12181 ctgattctgt ggataaccgt attaccgcta gatagctggt tgccctcgcc gctgggctgg
    12241 cggccgtcta tggccctgca aacgcgccag aaacgccgtc gaagccgtgt gcgagacacc
    12301 ccggccgccg gcgttgtgga tacctcgcgg aaaacttggc cctcactgac agatgagggg
    12361 cggacgttga cacttgaggg gccgactcac ccggcgcggc gttgacagat gaggggcagg
    12421 ctcgatttcg gccggcgacg tggagctggc cagcctcgca aatcggcgaa aacgcctgat
    12481 tttacgcgag tttcccacag atgatgtgga caagcctggg gataagtgcc ctgcggtatt
    12541 gacacttgag gggcgcgact actgacagat gaggggcgcg atccttgaca cttgaggggc
    12601 agagtgctga cagatgaggg gcgcacctat tgacatttga ggggctgtcc acaggcagaa
    12661 aatccagcat ttgcaagggt ttccgcccgt ttttcggcca ccgctaacct gtcttttaac
    12721 ctgcttttaa accaatattt ataaaccttg tttttaacca gggctgcgcc ctgtgcgcgt
    12781 gaccgcgcac gccgaagggg ggtgcccccc cttctcgaac cctcccggtc gagtcatggt
    12841 ctagcatgga tctcggggac gtcacccata atacccataa tagctgtttg ccagtacttt
    12901 gatcccgagg ggaaccctgt ggttggcatg cacatacaaa tggacgaacg gataaacctt
    12961 ttcacgccct tttaaatatc cgattattct aataaacgct cttttctctt aggtttaccc
    13021 gccaatatat cctgtcaaac actgatagtt taaactgaag gcgggaaacg acaatctgat
    13081 ccaagctcaa gctgctctag cattcgccat tcaggctgcg caactgttgg gaagggcgat
    13141 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
    13201 taagttgggt aacgccaggg ttttcccagt cacgacgtt
//
