LOCUS       sequence-320699-       16310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR plasmid with spCas9 expressed under egg cell promoter - GFP 
            seed selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16310)
  TITLE     CRISPR plasmid (spCas9 under EC1.2/EC1.1 promoter)
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 16310)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pEC1.2/EC1.1_spCas9_GFP
FEATURES             Location/Qualifiers
     source          1..16310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1362..1427
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1434..1454
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1479..5579
                     /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             1479..5579
                     /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             5492..5500
                     /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             5580..5627
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(6831..6833,6834..6836,6837..7547)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6831..6833,6834..6836,6837..7547)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6876..6897)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7137..7156)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7484..7505
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    7848..7872
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             8297..9091
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      9178..9766
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9667..9686
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9920..9937
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    9952..10092
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(10018..10037)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10178..10200
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      10436..10630
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(10696..11769)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(12198..12827)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    14128..14152
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     14340..14362
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     14354..14371
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     14355..14371
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             15084..15875
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_RNA        16027..16102
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 gaataaaagc atttgcgttt ggtttatcat tgcgtttata caaggacaga gatccactga
       61 gctggaatag cttaaaacca ttatcagaac aaaataaacc attttttgtt aagaatcaga
      121 gcatagtaaa caacagaaac aacctaagag aggtaacttg tccaagaaga tagctaatta
      181 tatctatttt ataaaagtta tcatagtttg taagtcacaa aagatgcaaa taacagagaa
      241 actaggagac ttgagaatat acattcttgt atatttgtat tcgagattgt gaaaatttga
      301 ccataagttt aaattcttaa aaagatatat ctgatctagg tgatggttat agactgtaat
      361 tttaccacat gtttaatgat ggatagtgac acacatgaca catcgacaac actatagcat
      421 cttatttaga ttacaacatg aaatttttct gtaatacatg tctttgtaca taatttaaaa
      481 gtaattccta agaaatatat ttatacaagg agtttaaaga aaacatagca taaagttcaa
      541 tgagtagtaa aaaccatata cagtatatag cataaagttc aatgagttta ttacaaaagc
      601 attggttcac tttctgtaac acgacgttaa accttcgtct ccaataggag cgctactgat
      661 tcaacatgcc aatatatact aaatacgttt ctacagtcaa atgctttaac gtttcatgat
      721 taagtgacta tttaccgtca atcctttccc attcctccca ctaatccaac tttttaatta
      781 ctcttaaatc accactaagc tagtaacgcc tatcatgaat tagctctact aaatctagca
      841 acctttcaaa tttgcagtat tgcaggtgtc tctgtgtctt taaaatagtt gccttatgat
      901 ttcttcggtt tcaagatgat caaatagtta tagatttcat gctcacacat gctcattaga
      961 tgtgtacata ctttacttac ccaaatctat tttctcgcaa agattttgat ggtaaagctg
     1021 atttggttct attgaactaa atcaaacgag tttcagactg agtgattcta atccggccca
     1081 ttagccccta aacagaccca ctaattacgc agcttttaat agagtaatta cacctagttt
     1141 acccactaaa ccactaagca ctaattatct cacaatctaa tgagcttccc tcgtaattac
     1201 ttgggctttc actctaccat ttatttgtaa cagtcaagtc tctactgtct ctatataaac
     1261 tctctaaagt taacacacaa ttctcatcac aaacaaatca accaaagcaa cttctactct
     1321 ttcttctttc gaccttatca atctgttgag aaatctagat ggattacaag gaccacgacg
     1381 gggattacaa ggaccacgac attgattaca aggatgatga tgacaagatg gctccgaaga
     1441 agaagaggaa ggttggcatc cacggggtgc cagctgctga caagaagtac tcgatcggcc
     1501 tcgatattgg gactaactct gttggctggg ccgtgatcac cgacgagtac aaggtgccct
     1561 caaagaagtt caaggtcctg ggcaacaccg atcggcattc catcaagaag aatctcattg
     1621 gcgctctcct gttcgacagc ggcgagacgg ctgaggctac gcggctcaag cgcaccgccc
     1681 gcaggcggta cacgcgcagg aagaatcgca tctgctacct gcaggagatt ttctccaacg
     1741 agatggcgaa ggttgacgat tctttcttcc acaggctgga ggagtcattc ctcgtggagg
     1801 aggataagaa gcacgagcgg catccaatct tcggcaacat tgtcgacgag gttgcctacc
     1861 acgagaagta ccctacgatc taccatctgc ggaagaagct cgtggactcc acagataagg
     1921 cggacctccg cctgatctac ctcgctctgg cccacatgat taagttcagg ggccatttcc
     1981 tgatcgaggg ggatctcaac ccggacaata gcgatgttga caagctgttc atccagctcg
     2041 tgcagacgta caaccagctc ttcgaggaga accccattaa tgcgtcaggc gtcgacgcga
     2101 aggctatcct gtccgctagg ctctcgaagt ctcggcgcct cgagaacctg atcgcccagc
     2161 tgccgggcga gaagaagaac ggcctgttcg ggaatctcat tgcgctcagc ctggggctca
     2221 cgcccaactt caagtcgaat ttcgatctcg ctgaggacgc caagctgcag ctctccaagg
     2281 acacatacga cgatgacctg gataacctcc tggcccagat cggcgatcag tacgcggacc
     2341 tgttcctcgc tgccaagaat ctgtcggacg ccatcctcct gtctgatatt ctcagggtga
     2401 acaccgagat tacgaaggct ccgctctcag cctccatgat caagcgctac gacgagcacc
     2461 atcaggatct gaccctcctg aaggcgctgg tcaggcagca gctccccgag aagtacaagg
     2521 agatcttctt cgatcagtcg aagaacggct acgctgggta cattgacggc ggggcctctc
     2581 aggaggagtt ctacaagttc atcaagccga ttctggagaa gatggacggc acggaggagc
     2641 tgctggtgaa gctcaatcgc gaggacctcc tgaggaagca gcggacattc gataacggca
     2701 gcatcccaca ccagattcat ctcggggagc tgcacgctat cctgaggagg caggaggact
     2761 tctacccttt cctcaaggat aaccgcgaga agatcgagaa gattctgact ttcaggatcc
     2821 cgtactacgt cggcccactc gctaggggca actcccgctt cgcttggatg acccgcaagt
     2881 cagaggagac gatcacgccg tggaacttcg aggaggtggt cgacaagggc gctagcgctc
     2941 agtcgttcat cgagaggatg acgaatttcg acaagaacct gccaaatgag aaggtgctcc
     3001 ctaagcactc gctcctgtac gagtacttca cagtctacaa cgagctgact aaggtgaagt
     3061 atgtgaccga gggcatgagg aagccggctt tcctgtctgg ggagcagaag aaggccatcg
     3121 tggacctcct gttcaagacc aaccggaagg tcacggttaa gcagctcaag gaggactact
     3181 tcaagaagat tgagtgcttc gattcggtcg agatctctgg cgttgaggac cgcttcaacg
     3241 cctccctggg gacctaccac gatctcctga agatcattaa ggataaggac ttcctggaca
     3301 acgaggagaa tgaggatatc ctcgaggaca ttgtgctgac actcactctg ttcgaggacc
     3361 gggagatgat cgaggagcgc ctgaagactt acgcccatct cttcgatgac aaggtcatga
     3421 agcagctcaa gaggaggagg tacaccggct gggggaggct gagcaggaag ctcatcaacg
     3481 gcattcggga caagcagtcc gggaagacga tcctcgactt cctgaagagc gatggcttcg
     3541 cgaaccgcaa tttcatgcag ctgattcacg atgacagcct cacattcaag gaggatatcc
     3601 agaaggctca ggtgagcggc cagggggact cgctgcacga gcatatcgcg aacctcgctg
     3661 gctcgccagc tatcaagaag gggattctgc agaccgtgaa ggttgtggac gagctggtga
     3721 aggtcatggg caggcacaag cctgagaaca tcgtcattga gatggcccgg gagaatcaga
     3781 ccacgcagaa gggccagaag aactcacgcg agaggatgaa gaggatcgag gagggcatta
     3841 aggagctggg gtcccagatc ctcaaggagc acccggtgga gaacacgcag ctgcagaatg
     3901 agaagctcta cctgtactac ctccagaatg gccgcgatat gtatgtggac caggagctgg
     3961 atattaacag gctcagcgat tacgacgtcg atcatatcgt tccacagtca ttcctgaagg
     4021 atgactccat tgacaacaag gtcctcacca ggtcggacaa gaaccggggc aagtctgata
     4081 atgttccttc agaggaggtc gttaagaaga tgaagaacta ctggcgccag ctcctgaatg
     4141 ccaagctgat cacgcagcgg aagttcgata acctcacaaa ggctgagagg ggcgggctct
     4201 ctgagctgga caaggcgggc ttcatcaaga ggcagctggt cgagacacgg cagatcacta
     4261 agcacgttgc gcagattctc gactcacgga tgaacactaa gtacgatgag aatgacaagc
     4321 tgatccgcga ggtgaaggtc atcaccctga agtcaaagct cgtctccgac ttcaggaagg
     4381 atttccagtt ctacaaggtt cgggagatca acaattacca ccatgcccat gacgcgtacc
     4441 tgaacgcggt ggtcggcaca gctctgatca agaagtaccc aaagctcgag agcgagttcg
     4501 tgtacgggga ctacaaggtt tacgatgtga ggaagatgat cgccaagtcg gagcaggaga
     4561 ttggcaaggc taccgccaag tacttcttct actctaacat tatgaatttc ttcaagacag
     4621 agatcactct ggccaatggc gagatccgga agcgccccct catcgagacg aacggcgaga
     4681 cgggggagat cgtgtgggac aagggcaggg atttcgcgac cgtcaggaag gttctctcca
     4741 tgccacaagt gaatatcgtc aagaagacag aggtccagac tggcgggttc tctaaggagt
     4801 caattctgcc taagcggaac agcgacaagc tcatcgcccg caagaaggac tgggatccga
     4861 agaagtacgg cgggttcgac agccccactg tggcctactc ggtcctggtt gtggcgaagg
     4921 ttgagaaggg caagtccaag aagctcaaga gcgtgaagga gctgctgggg atcacgatta
     4981 tggagcgctc cagcttcgag aagaacccga tcgatttcct ggaggcgaag ggctacaagg
     5041 aggtgaagaa ggacctgatc attaagctcc ccaagtactc actcttcgag ctggagaacg
     5101 gcaggaagcg gatgctggct tccgctggcg agctgcagaa ggggaacgag ctggctctgc
     5161 cgtccaagta tgtgaacttc ctctacctgg cctcccacta cgagaagctc aagggcagcc
     5221 ccgaggacaa cgagcagaag cagctgttcg tcgagcagca caagcattac ctcgacgaga
     5281 tcattgagca gatttccgag ttctccaagc gcgtgatcct ggccgacgcg aatctggata
     5341 aggtcctctc cgcgtacaac aagcaccgcg acaagccaat cagggagcag gctgagaata
     5401 tcattcatct cttcaccctg acgaacctcg gcgcccctgc tgctttcaag tacttcgaca
     5461 caactatcga tcgcaagagg tacacaagca ctaaggaggt cctggacgcg accctcatcc
     5521 accagtcgat taccggcctc tacgagacgc gcatcgacct gtctcagctc gggggcgaca
     5581 agcggccagc ggcgacgaag aaggcggggc aggcgaagaa gaagaagtga gctcagagct
     5641 ttcgttcgta tcatcggttt cgacaacgtt cgtcaagttc aatgcatcag tttcattgcg
     5701 cacacaccag aatcctactg agtttgagta ttatggcatt gggaaaactg tttttcttgt
     5761 accatttgtt gtgcttgtaa tttactgtgt tttttattcg gttttcgcta tcgaactgtg
     5821 aaatggaaat ggatggagaa gagttaatga atgatatggt ccttttgttc attctcaaat
     5881 taatattatt tgttttttct cttatttgtt gtgtgttgaa tttgaaatta taagagatat
     5941 gcaaacattt tgttttgagt aaaaatgtgt caaatcgtgg cctctaatga ccgaagttaa
     6001 tatgaggagt aaaacacttg tagttgtacc attatgctta ttcactaggc aacaaatata
     6061 ttttcagacc tagaaaagct gcaaatgtta ctgaatacaa gtatgtcctc ttgtgtttta
     6121 gacatttatg aactttcctt tatgtaattt tccagaatcc ttgtcagatt ctaatcattg
     6181 ctttataatt atagttatac tcatggattt gtagttgagt atgaaaatat tttttaatgc
     6241 attttatgac ttgccaattg attgacaacg aattggaaac caaattaaca tagggttttt
     6301 atttaaataa aagtttaacc ttctttttaa aaaattgttc atagtgtcat gtcagaacaa
     6361 gagctacaaa tcacacatag catgcataag cggagctatg atgagtggta ttgttttgtt
     6421 cgtcacttgt cactcttttc caacacataa tcccgacaac aacgtaagag catctctctc
     6481 tctccacaca cactcatgca tgcatgcatt cttacacgtg attgccatgc aaatctcctt
     6541 tctcacctat aaatacaaac caacccttca ctacactctt cactcaaacc aaaacaagaa
     6601 aacatacaca aatagcaaaa caagggcgaa ttctcaacac aacatataca aaacaaacga
     6661 atctcaagca atcaagcatt ctacttctat tgcagcaatt taaatcattt cttttaaagc
     6721 aaaagcaatt ttctgaaaat tttcaccatt tacgaacgat agccatggtg aagactaatc
     6781 tttttctctt tctcatcttt tcacttctcc tatcattatc ctcggccgac atggtgagca
     6841 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     6901 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     6961 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     7021 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     7081 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     7141 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     7201 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     7261 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     7321 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     7381 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     7441 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     7501 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagaag gacgagctgt
     7561 aagcgccgcc cggctgcatc gctgaaatca ccagtctctc tctacaaatc tatctctctc
     7621 tataataatg tgtgagtagt tcccagataa gggaattagg gttcttatag ggtttcgctc
     7681 atgtgttgag catataagaa acccttagta tgtatttgta tttgtaaaat acttctatca
     7741 ataaaatttc taattcctaa aaccaaaatc cagtggatta attcggcgtt aattcagtac
     7801 attaaaaacg tccgcaatgt gttattaagt tgtctaagcg tcaatttgtt tacaccacaa
     7861 tatatcctgc caccagccag ccaacagctc cccgaccggc agctcggcac aaaatcacca
     7921 ctcgatacag gcagcccatc agtccgggac ggcgtcagcg ggagagccgt tgtaaggcgg
     7981 cagactttgc tcatgttacc gatgctattc ggaagaacgg caactaagct gccgggtttg
     8041 aaacacggat gatctcgcgg agggtagcat gttgattgta acgatgacag agcgttgctg
     8101 cctgtgatca ccgcggtttc aaaatcggct ccgtcgatac tatgttatac gccaactttg
     8161 aaaacaactt tgaaaaagct gttttctggt atttaaggtt ttagaatgca aggaacagtg
     8221 aattggagtt cgtcttgtta taattagctt cttggggtat ctttaaatac tgtagaaaag
     8281 aggaaggaaa taataaatgg ctaaaatgag aatatcaccg gaattgaaaa aactgatcga
     8341 aaaataccgc tgcgtaaaag atacggaagg aatgtctcct gctaaggtat ataagctggt
     8401 gggagaaaat gaaaacctat atttaaaaat gacggacagc cggtataaag ggaccaccta
     8461 tgatgtggaa cgggaaaagg acatgatgct atggctggaa ggaaagctgc ctgttccaaa
     8521 ggtcctgcac tttgaacggc atgatggctg gagcaatctg ctcatgagtg aggccgatgg
     8581 cgtcctttgc tcggaagagt atgaagatga acaaagccct gaaaagatta tcgagctgta
     8641 tgcggagtgc atcaggctct ttcactccat cgacatatcg gattgtccct atacgaatag
     8701 cttagacagc cgcttagccg aattggatta cttactgaat aacgatctgg ccgatgtgga
     8761 ttgcgaaaac tgggaagaag acactccatt taaagatccg cgcgagctgt atgatttttt
     8821 aaagacggaa aagcccgaag aggaacttgt cttttcccac ggcgacctgg gagacagcaa
     8881 catctttgtg aaagatggca aagtaagtgg ctttattgat cttgggagaa gcggcagggc
     8941 ggacaagtgg tatgacattg ccttctgcgt ccggtcgatc agggaggata tcggggaaga
     9001 acagtatgtc gagctatttt ttgacttact ggggatcaag cctgattggg agaaaataaa
     9061 atattatatt ttactggatg aattgtttta gtacctagaa tgcatgacca aaatccctta
     9121 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     9181 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     9241 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     9301 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     9361 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     9421 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     9481 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     9541 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     9601 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     9661 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     9721 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     9781 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     9841 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     9901 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     9961 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
    10021 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
    10081 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
    10141 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
    10201 ttttcaccgt catcaccgaa acgcgcgagg cagggtgcct tgatgtgggc gccggcggtc
    10261 gagtggcgac ggcgcggctt gtccgcgccc tggtagattg cctggccgta ggccagccat
    10321 ttttgagcgg ccagcggccg cgataggccg acgcgaagcg gcggggcgta gggagcgcag
    10381 cgaccgaagg gtaggcgctt tttgcagctc ttcggctgtg cgctggccag acagttatgc
    10441 acaggccagg cgggttttaa gagttttaat aagttttaaa gagttttagg cggaaaaatc
    10501 gccttttttc tcttttatat cagtcactta catgtgtgac cggttcccaa tgtacggctt
    10561 tgggttccca atgtacgggt tccggttccc aatgtacggc tttgggttcc caatgtacgt
    10621 gctatccaca ggaaacagac cttttcgacc tttttcccct gctagggcaa tttgccctag
    10681 catctgctcc gtacattagg aaccggcgga tgcttcgccc tcgatcaggt tgcggtagcg
    10741 catgactagg atcgggccag cctgccccgc ctcctccttc aaatcgtact ccggcaggtc
    10801 atttgacccg atcagcttgc gcacggtgaa acagaacttc ttgaactctc cggcgctgcc
    10861 actgcgttcg tagatcgtct tgaacaacca tctggcttct gccttgcctg cggcgcggcg
    10921 tgccaggcgg tagagaaaac ggccgatgcc gggatcgatc aaaaagtaat cggggtgaac
    10981 cgtcagcacg tccgggttct tgccttctgt gatctcgcgg tacatccaat cagctagctc
    11041 gatctcgatg tactccggcc gcccggtttc gctctttacg atcttgtagc ggctaatcaa
    11101 ggcttcaccc tcggataccg tcaccaggcg gccgttcttg gccttcttcg tacgctgcat
    11161 ggcaacgtgc gtggtgttta accgaatgca ggtttctacc aggtcgtctt tctgctttcc
    11221 gccatcggct cgccggcaga acttgagtac gtccgcaacg tgtggacgga acacgcggcc
    11281 gggcttgtct cccttccctt cccggtatcg gttcatggat tcggttagat gggaaaccgc
    11341 catcagtacc aggtcgtaat cccacacact ggccatgccg gccggccctg cggaaacctc
    11401 tacgtgcccg tctggaagct cgtagcggat cacctcgcca gctcgtcggt cacgcttcga
    11461 cagacggaaa acggccacgt ccatgatgct gcgactatcg cgggtgccca cgtcatagag
    11521 catcggaacg aaaaaatctg gttgctcgtc gcccttgggc ggcttcctaa tcgacggcgc
    11581 accggctgcc ggcggttgcc gggattcttt gcggattcga tcagcggccg cttgccacga
    11641 ttcaccgggg cgtgcttctg cctcgatgcg ttgccgctgg gcggcctgcg cggccttcaa
    11701 cttctccacc aggtcatcac ccagcgccgc gccgatttgt accgggccgg atggtttgcg
    11761 accgctcacg ccgattcctc gggcttgggg gttccagtgc cattgcaggg ccggcaggca
    11821 acccagccgc ttacgcctgg ccaaccgccc gttcctccac acatggggca ttccacggcg
    11881 tcggtgcctg gttgttcttg attttccatg ccgcctcctt tagccgctaa aattcatcta
    11941 ctcatttatt catttgctca tttactctgg tagctgcgcg atgtattcag atagcagctc
    12001 ggtaatggtc ttgccttggc gtaccgcgta catcttcagc ttggtgtgat cctccgccgg
    12061 caactgaaag ttgacccgct tcatggctgg cgtgtctgcc aggctggcca acgttgcagc
    12121 cttgctgctg cgtgcgctcg gacggccggc acttagcgtg tttgtgcttt tgctcatttt
    12181 ctctttacct cattaactca aatgagtttt gatttaattt cagcggccag cgcctggacc
    12241 tcgcgggcag cgtcgccctc gggttctgat tcaagaacgg ttgtgccggc ggcggcagtg
    12301 cctgggtagc tcacgcgctg cgtgatacgg gactcaagaa tgggcagctc gtacccggcc
    12361 agcgcctcgg caacctcacc gccgatgcgc gtgcctttga tcgcccgcga cacgacaaag
    12421 gccgcttgta gccttccatc cgtgacctca atgcgctgct taaccagctc caccaggtcg
    12481 gcggtggccc atatgtcgta agggcttggc tgcaccggaa tcagcacgaa gtcggctgcc
    12541 ttgatcgcgg acacagccaa gtccgccgcc tggggcgctc cgtcgatcac tacgaagtcg
    12601 cgccggccga tggccttcac gtcgcggtca atcgtcgggc ggtcgatgcc gacaacggtt
    12661 agcggttgat cttcccgcac ggccgcccaa tcgcgggcac tgccctgggg atcggaatcg
    12721 actaacagaa catcggcccc ggcgagttgc agggcgcggg ctagatgggt tgcgatggtc
    12781 gtcttgcctg acccgccttt ctggttaagt acagcgataa ccttcatgcg ttccccttgc
    12841 gtatttgttt atttactcat cgcatcatat acgcagcgac cgcatgacgc aagctgtttt
    12901 actcaaatac acatcacctt tttagacggc ggcgctcggt ttcttcagcg gccaagctgg
    12961 ccggccaggc cgccagcttg gcatcagaca aaccggccag gatttcatgc agccgcacgg
    13021 ttgagacgtg cgcgggcggc tcgaacacgt acccggccgc gatcatctcc gcctcgatct
    13081 cttcggtaat gaaaaacggt tcgtcctggc cgtcctggtg cggtttcatg cttgttcctc
    13141 ttggcgttca ttctcggcgg ccgccagggc gtcggcctcg gtcaatgcgt cctcacggaa
    13201 ggcaccgcgc cgcctggcct cggtgggcgt cacttcctcg ctgcgctcaa gtgcgcggta
    13261 cagggtcgag cgatgcacgc caagcagtgc agccgcctct ttcacggtgc ggccttcctg
    13321 gtcgatcagc tcgcgggcgt gcgcgatctg tgccggggtg agggtagggc gggggccaaa
    13381 cttcacgcct cgggccttgg cggcctcgcg cccgctccgg gtgcggtcga tgattaggga
    13441 acgctcgaac tcggcaatgc cggcgaacac ggtcaacacc atgcggccgg ccggcgtggt
    13501 ggtgtcggcc cacggctctg ccaggctacg caggcccgcg ccggcctcct ggatgcgctc
    13561 ggcaatgtcc agtaggtcgc gggtgctgcg ggccaggcgg tctagcctgg tcactgtcac
    13621 aacgtcgcca gggcgtaggt ggtcaagcat cctggccagc tccgggcggt cgcgcctggt
    13681 gccggtgatc ttctcggaaa acagcttggt gcagccggcc gcgtgcagtt cggcccgttg
    13741 gttggtcaag tcctggtcgt cggtgctgac gcgggcatag cccagcaggc cagcggcggc
    13801 gctcttgttc atggcgtaat gtctccggtt ctagtcgcaa gtattctact ttatgcgact
    13861 aaaacacgcg acaagaaaac gccaggaaaa gggcagggcg gcagcctgtc gcgtaactta
    13921 ggacttgtgc gacatgtcgt tttcagaaga cggctgcact gaacgtcaga agccgactgc
    13981 actatagcag cggaggggtt ggatcaaagt actttgatcc cgaggggaac cctgtggttg
    14041 gcatgcacat acaaatggac gaacggataa accttttcac gcccttttaa atatccgatt
    14101 attctaataa acgctctttt ctcttaggtt tacccgccaa tatatcctgt caaacactga
    14161 tagtttaaac tgaaggcggg aaacgacaat ctgatccaag ctcaagctgc tctagcattc
    14221 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
    14281 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
    14341 ccagtcacga cgttgtaaaa cgacggccag tgccaagctt cgacttgcct tccgcacaat
    14401 acatcatttc ttcttagctt tttttcttct tcttcgttca tacagttttt ttttgtttat
    14461 cagcttacat tttcttgaac cgtagctttc gttttcttct ttttaacttt ccattcggag
    14521 tttttgtatc ttgtttcata gtttgtccca ggattagaat gattaggcat cgaaccttca
    14581 agaatttgat tgaataaaac atcttcattc ttaagatatg aagataatct tcaaaaggcc
    14641 cctgggaatc tgaaagaaga gaagcaggcc catttatatg ggaaagaaca atagtatttc
    14701 ttatataggc ccatttaagt tgaaaacaat cttcaaaagt cccacatcgc ttagataaga
    14761 aaacgaagct gagtttatat acagctagag tcgaagtagt gattgggaga ccaacccagt
    14821 ggacataagc ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact
    14881 ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt
    14941 gttatgactg tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt
    15001 acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc
    15061 gccctaaaac aaagttaaac atcatggggg aagcggtgat cgccgaagta tcgactcaac
    15121 tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt
    15181 tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta
    15241 cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa
    15301 cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc
    15361 acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc
    15421 agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta
    15481 tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac
    15541 tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat
    15601 ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca
    15661 tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa
    15721 tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg
    15781 gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg
    15841 tgaaaggcga gatcaccaag gtagtcggca aataatgtct agctagaaat tcgttcaagc
    15901 cgacgccgct tcgcggcgcg gcttaactca agcgttagat gcactaagca cataattgct
    15961 cacagccaaa ctatcaggtc aagtctgctt ttattatttt taagcgtgca taataagccg
    16021 gtctcggttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
    16081 aaaagtggca ccgagtcggt gctttttttt gcaaaatttt ccagatcgat ttcttcttcc
    16141 tctgttcttc ggcgttcaat ttctggggtt ttctcttcgt tttctgtaac tgaaacctaa
    16201 aatttgacct aaaaaaaatc tcaaataata tgattcagtg gttttgtact tttcagttag
    16261 ttgagttttg cagttccgat gagataaacc aataccatgg ttatactagt
//
