LOCUS       sequence-220965-       13094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway destination vector encoding Maize ubiquitin promoter driving
            Cas9 and 35S promoter driving aminoglycoside phosphotransferase for 
            G418 selection in plants..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13094)
  AUTHORS   Mallett DR, Chang M, Cheng X, Bezanilla M
  TITLE     Efficient and modular CRISPR-Cas9 vector system for Physcomitrella 
            patens.
  JOURNAL   Plant Direct. 2019 Sep 12;3(9):e00168. doi: 10.1002/pld3.168. 
            eCollection 2019 Sep.
  PUBMED    31523744
REFERENCE   2  (bases 1 to 13094)
  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 
            Direct."; date: "2019-09-12"; volume: "3(9):e00168. doi"; pages: " 
            10.1002/pld3.168. eCollection 2019 Sep"
COMMENT     Sequence Label: pMK-Cas9-gate
FEATURES             Location/Qualifiers
     source          1..13094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    49..173
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(198..203,204..221,222..228)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             282..941
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(348..367)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1283..1588
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1467..1486
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(1629..1753)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1776..3768
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     protein_bind    3791..3815
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3842..3862
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3863..7966
                     /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             3863..7963
                     /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"
     protein_bind    complement(7983..8007)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      8020..8029
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        8427..8606
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      8672..8924
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        8957..9302
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     9245..9265
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             9332..10126
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(9386..9405)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9996..10015
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    10196..10372
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(10923..10939)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10923..10939)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10936..10958)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10947..10963
                     /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(10971..10977,10978..10995,10996..11001))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11016..11037
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11154..11171)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11325..11913)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11405..11424)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12084..12875,12876..12944))
                     /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     12707..12726
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12945..13049)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaactgaagg cgggaaacga caatctgatc caagctcaag ctaagcttac aagtttgtac
       61 aaaaaagctg aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg
      121 cataaaaaac agactacata atactgtaaa acacaacata tccagtcact atggcggccg
      181 cattaggcac cccaggcttt acactttatg cttccggctc gtataatgtg tggattttga
      241 gttaggatcc gtcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
      301 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
      361 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
      421 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
      481 tgaatgctca tccggaattc cgtatggcaa tgaaagacgg tgagctggtg atatgggata
      541 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga
      601 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt
      661 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag
      721 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct
      781 tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc
      841 tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg
      901 aattacaaca gtactgcgat gagtggcagg gcggggcgta aacgcgtgga tccggcttac
      961 taaaagccag ataacagtat gcgtatttgc gcgctgattt ttgcggtata agaatatata
     1021 ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct atgaagcagc gtattacagt
     1081 gacagttgac agcgacagct atcagttgct caaggcatat atgatgtcaa tatctccggt
     1141 ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc gtgccgaacg ctggaaagcg
     1201 gaaaatcagg aagggatggc tgaggtcgcc cggtttattg aaatgaacgg ctcttttgct
     1261 gacgagaaca ggggctggtg aaatgcagtt taaggtttac acctataaaa gagagagccg
     1321 ttatcgtctg tttgtggatg tacagagtga tattattgac acgcccgggc gacggatggt
     1381 gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac tttacccggt
     1441 ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca gtgtgccggt
     1501 ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc
     1561 cattaacctg atgttctggg gaatataaat gtcaggctcc cttatacaca gccagtctgc
     1621 aggtcgacca tagtgactgg atatgttgtg ttttacagta ttatgtagtc tgttttttat
     1681 gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt tcagctttct
     1741 tgtacaaagt ggttctagag atcaagcttg catgcctgca gtgcagcgtg acccggtcgt
     1801 gcccctctct agagataatg agcattgcat gtctaagtta taaaaaatta ccacatattt
     1861 tttttgtcac acttgtttga agtgcagttt atctatcttt atacatatat ttaaacttta
     1921 ctctacgaat aatataatct atagtactac aataatatca gtgttttaga gaatcatata
     1981 aatgaacagt tagacatggt ctaaaggaca attgagtatt ttgacaacag gactctacag
     2041 ttttatcttt ttagtgtgca tgtgttctcc tttttttttg caaatagctt cacctatata
     2101 atacttcatc cattttatta gtacatccat ttagggttta gggttaatgg tttttataga
     2161 ctaatttttt tagtacatct attttattct attttagcct ctaaattaag aaaactaaaa
     2221 ctctatttta gtttttttat ttaataattt agatataaaa tagaataaaa taaagtgact
     2281 aaaaattaaa caaataccct ttaagaaatt aaaaaaacta aggaaacatt tttcttgttt
     2341 cgagtagata atgccagcct gttaaacgcc gtcgacgagt ctaacggaca ccaaccagcg
     2401 aaccagcagc gtcgcgtcgg gccaagcgaa gcagacggca cggcatctct gtcgctgcct
     2461 ctggacccct ctcgagagtt ccgctccacc gttggacttg ctccgctgtc ggcatccaga
     2521 aattgcgtgg cggagcggca gacgtgagcc ggcacggcag gcggcctcct cctcctctca
     2581 cggcaccggc agctacgggg gattcctttc ccaccgctcc ttcgctttcc cttcctcgcc
     2641 cgccgtaata aatagacacc ccctccacac cctctttccc caacctcgtg ttgttcggag
     2701 cgcacacaca cacaaccaga tctcccccaa atccacccgt cggcacctcc gcttcaaggt
     2761 acgccgctcg tcctcccccc ccccccctct ctaccttctc tagatcggcg ttccggtcca
     2821 tggttagggc ccggtagttc tacttctgtt catgtttgtg ttagatccgt gtttgtgtta
     2881 gatccgtgct gctagcgttc gtacacggat gcgacctgta cgtcagacac gttctgattg
     2941 ctaacttgcc agtgtttctc tttggggaat cctgggatgg ctctagccgt tccgcagacg
     3001 ggatcgattt catgattttt tttgtttcgt tgcatagggt ttggtttgcc cttttccttt
     3061 atttcaatat atgccgtgca cttgtttgtc gggtcatctt ttcatgcttt tttttgtctt
     3121 ggttgtgatg atgtggtctg gttgggcggt cgttctagat cggagtagaa ttctgtttca
     3181 aactacctgg tggatttatt aattttggat ctgtatgtgt gtgccataca tattcatagt
     3241 tacgaattga agatgatgga tggaaatatc gatctaggat aggtatacat gttgatgcgg
     3301 gttttactga tgcatataca gagatgcttt ttgttcgctt ggttgtgatg atgtggtgtg
     3361 gttgggcggt cgttcattcg ttctagatcg gagtagaata ctgtttcaaa ctacctggtg
     3421 tatttattaa ttttggaact gtatgtgtgt gtcatacatc ttcatagtta cgagtttaag
     3481 atggatggaa atatcgatct aggataggta tacatgttga tgtgggtttt actgatgcat
     3541 atacatgatg gcatatgcag catctattca tatgctctaa ccttgagtac ctatctatta
     3601 taataaacaa gtatgtttta taattatttt gatcttgata tacttggatg atggcatatg
     3661 cagcagctat atgtggattt ttttagccct gccttcatac gctatttatt tgcttggtac
     3721 tgtttctttt gtcgatgctc accctgttgt ttggtgttac ttctgcaggt cgactctaga
     3781 ggatcccggg gcaagtttgt acaaaaaagc aggctccgcg gccgccccct tcaccatggc
     3841 cccaaagaag aagcgcaagg tcgacaagaa gtactccatc ggcctcgaca tcggcaccaa
     3901 ttctgttggc tgggccgtga tcaccgacga gtacaaggtg ccgtccaaga agttcaaggt
     3961 cctcggcaac accgaccgcc actccatcaa gaagaatctc atcggcgccc tgctgttcga
     4021 ctctggcgag acagccgagg ctacaaggct caagaggacc gctagacgca ggtacaccag
     4081 gcgcaagaac cgcatctgct acctccaaga gatcttctcc aacgagatgg ccaaggtgga
     4141 cgacagcttc ttccacaggc tcgaggagag cttcctcgtc gaggaggaca agaagcacga
     4201 gcgccatccg atcttcggca acatcgtgga tgaggtggcc taccacgaga agtacccgac
     4261 catctaccac ctccgcaaga agctcgtcga ctccaccgat aaggccgacc tcaggctcat
     4321 ctacctcgcc ctcgcccaca tgatcaagtt caggggccac ttcctcatcg agggcgacct
     4381 caacccggac aactccgatg tggacaagct gttcatccag ctcgtgcaga cctacaacca
     4441 gctgttcgag gagaacccga tcaacgcctc tggcgttgac gccaaggcta ttctctctgc
     4501 caggctctct aagtcccgca ggctcgagaa tctgatcgcc caacttccgg gcgagaagaa
     4561 gaatggcctc ttcggcaacc tgatcgccct ctctcttggc ctcaccccga acttcaagtc
     4621 caacttcgac ctcgccgagg acgccaagct ccagctttcc aaggacacct acgacgacga
     4681 cctcgacaat ctcctcgccc agattggcga tcagtacgcc gatctgttcc tcgccgccaa
     4741 gaatctctcc gacgccatcc tcctcagcga catcctcagg gtgaacaccg agatcaccaa
     4801 ggccccactc tccgcctcca tgatcaagag gtacgacgag caccaccagg acctcacact
     4861 cctcaaggcc ctcgtgagac agcagctccc agagaagtac aaggagatct tcttcgacca
     4921 gtccaagaac ggctacgccg gctacatcga tggcggcgct tctcaagagg agttctacaa
     4981 gttcatcaag ccgatcctcg agaagatgga cggcaccgag gagctgctcg tgaagctcaa
     5041 tagagaggac ctcctccgca agcagcgcac cttcgataat ggctccatcc cgcaccagat
     5101 ccacctcggc gagcttcatg ctatcctccg caggcaagag gacttctacc cgttcctcaa
     5161 ggacaaccgc gagaagattg agaagatcct caccttccgc atcccgtact acgtgggccc
     5221 gctcgccagg ggcaactcca ggttcgcctg gatgaccaga aagtccgagg agacaatcac
     5281 cccctggaac ttcgaggagg tggtggataa gggcgcctct gcccagtctt tcatcgagcg
     5341 catgaccaac ttcgacaaga acctcccgaa cgagaaggtg ctcccgaagc actcactcct
     5401 ctacgagtac ttcaccgtgt acaacgagct gaccaaggtg aagtacgtga ccgaggggat
     5461 gaggaagcca gctttcctta gcggcgagca aaagaaggcc atcgtcgacc tgctgttcaa
     5521 gaccaaccgc aaggtgaccg tgaagcagct caaggaggac tacttcaaga aaatcgagtg
     5581 cttcgactcc gtcgagatct ccggcgtcga ggataggttc aatgcctccc tcgggaccta
     5641 ccacgacctc ctcaagatta tcaaggacaa ggacttcctc gacaacgagg agaacgagga
     5701 catcctcgag gacatcgtgc tcaccctcac cctcttcgag gaccgcgaga tgatcgagga
     5761 gcgcctcaag acatacgccc acctcttcga cgacaaggtg atgaagcagc tgaagcgcag
     5821 gcgctatacc ggctggggca ggctctctag gaagctcatc aacggcatcc gcgacaagca
     5881 gtccggcaag acgatcctcg acttcctcaa gtccgacggc ttcgccaacc gcaacttcat
     5941 gcagctcatc cacgacgact ccctcacctt caaggaggac atccaaaagg cccaggtgtc
     6001 cggccaaggc gattccctcc atgaacatat cgccaatctc gccggctccc cggctatcaa
     6061 gaagggcatt ctccagaccg tgaaggtggt ggacgagctg gtgaaggtga tgggcaggca
     6121 caagccagag aacatcgtga tcgagatggc ccgcgagaac cagaccacac agaagggcca
     6181 aaagaactcc cgcgagcgca tgaagaggat cgaggagggc attaaggagc tgggctccca
     6241 gatcctcaag gagcacccag tcgagaacac ccagctccag aacgagaagc tctacctcta
     6301 ctacctccag aacggccgcg acatgtacgt ggaccaagag ctggacatca accgcctctc
     6361 cgactacgac gtggaccata ttgtgccgca gtccttcctg aaggacgact ccatcgacaa
     6421 caaggtgctc acccgctccg acaagaacag gggcaagtcc gataacgtgc cgtccgaaga
     6481 ggtcgtcaag aagatgaaga actactggcg ccagctcctc aacgccaagc tcatcaccca
     6541 gaggaagttc gacaacctca ccaaggccga gagaggcggc ctttccgagc ttgataaggc
     6601 cggcttcatc aagcgccagc tcgtcgagac acgccagatc acaaagcacg tggcccagat
     6661 cctcgactcc cgcatgaaca ccaagtacga cgagaacgac aagctcatcc gcgaggtgaa
     6721 ggtcatcacc ctcaagtcca agctcgtgtc cgacttccgc aaggacttcc agttctacaa
     6781 ggtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctcaatg ccgtggtggg
     6841 cacagccctc atcaagaagt acccaaagct cgagtccgag ttcgtgtacg gcgactacaa
     6901 ggtgtacgac gtgcgcaaga tgatcgccaa gtccgagcaa gagatcggca aggcgaccgc
     6961 caagtacttc ttctactcca acatcatgaa tttcttcaag accgagatca cgctcgccaa
     7021 cggcgagatt aggaagaggc cgctcatcga gacaaacggc gagacaggcg agatcgtgtg
     7081 ggacaagggc agggatttcg ccacagtgcg caaggtgctc tccatgccgc aagtgaacat
     7141 cgtgaagaag accgaggttc agaccggcgg cttctccaag gagtccatcc tcccaaagcg
     7201 caactccgac aagctgatcg cccgcaagaa ggactgggac ccgaagaagt atggcggctt
     7261 cgattctccg accgtggcct actctgtgct cgtggttgcc aaggtcgaga agggcaagag
     7321 caagaagctc aagtccgtca aggagctgct gggcatcacg atcatggagc gcagcagctt
     7381 cgagaagaac ccaatcgact tcctcgaggc caagggctac aaggaggtga agaaggacct
     7441 catcatcaag ctcccgaagt acagcctctt cgagcttgag aacggccgca agagaatgct
     7501 cgcctctgct ggcgagcttc agaagggcaa cgagcttgct ctcccgtcca agtacgtgaa
     7561 cttcctctac ctcgcctccc actacgagaa gctcaagggc tccccagagg acaacgagca
     7621 aaagcagctg ttcgtcgagc agcacaagca ctacctcgac gagatcatcg agcagatctc
     7681 cgagttctcc aagcgcgtga tcctcgccga tgccaacctc gataaggtgc tcagcgccta
     7741 caacaagcac cgcgataagc caattcgcga gcaggccgag aacatcatcc acctcttcac
     7801 cctcaccaac ctcggcgctc cagccgcctt caagtacttc gacaccacca tcgaccgcaa
     7861 gcgctacacc tctaccaagg aggttctcga cgccaccctc atccaccagt ctatcacagg
     7921 cctctacgag acacgcatcg acctctcaca actcggcggc gattgaaagg gtgggcgcgc
     7981 cgacccagct ttcttgtaca aagtggtgat atcccgcggc catggcggcc gggagcatgc
     8041 gacgtcgatc taactgacta gccgcggcca tgctagagtc cgcaaaaatc accagtctct
     8101 ctctacaaat ctatctctct ctatttttct ccagaataat gtgtgagtag ttcccagata
     8161 agggaattag ggttcttata gggtttcgct catgtgttga gcatataaga aacccttagt
     8221 atgtatttgt atttgtaaaa tacttctatc aataaaattt ctaattccta aaaccaaaat
     8281 ccagtgacct gcaggcatgc gacgtcgggc cctctagagg atccccgggt accgcgaatt
     8341 atcgatcatg agcggagaat taagggagtc acgttatgac ccccgccgat gacgcgggac
     8401 aagccgtttt acgtttggaa ctgacagaac cgcaacgttg aaggagccac tgagccgcgg
     8461 gtttctggag tttaatgagc taagcacata cgtcagaaac cattattgcg cgttcaaaag
     8521 tcgcctaagg tcactatcag ctagcaaata tttcttgtca aaaatgctcc actgacgttc
     8581 cataaattcc cctcggtatc caattagagt ctcatattca ctctcaactc gaatcccccc
     8641 tatgaaaaag cctgaactca ccgcgacgtc cgatcgttca aacatttggc aataaagttt
     8701 cttaagattg aatcctgttg ccggtcttgc gatgattatc atataatttc tgttgaatta
     8761 cgttaagcat gtaataatta acatgtaatg catgacgtta tttatgagat gggtttttat
     8821 gattagagtc ccgcaattat acatttaata cgcgatagaa aacaaaatat agcgcgcaaa
     8881 ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta gatcggggat acagtctcag
     8941 aagaccaaag ggctattgag acttttcaac aaagggtaat ttcgggaaac ctcctcggat
     9001 tccattgccc agctatctgt cacttcatcg aaaggacagt agaaaaggaa ggtggctcct
     9061 acaaatgcca tcattgcgat aaaggaaagg ctatcattca agatgcctct gccgacagtg
     9121 gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca
     9181 cgtcttcaaa gcaagtggat tgatgtgaca tctccactga cgtaagggat gacgcacaat
     9241 cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga
     9301 cagcccaagc atggagaacc gacctgcagg catgattgaa caagatggat tgcacgcagg
     9361 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     9421 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     9481 gaccgacctg tccggtgccc tgaatgaact ccaagacgag gcagcgcggc tatcgtggct
     9541 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     9601 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     9661 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     9721 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     9781 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     9841 gttcgccagg ctcaaggcgc ggatgcccga cggcgaggat ctcgtcgtga cccatggcga
     9901 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     9961 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
    10021 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
    10081 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
    10141 ttcggacggt acgctgaaat caccagtctc tctctacaaa tctatctctc tctattttct
    10201 ccataaataa tgtgtgagta gtttcccgat aagggaaatt agggttctta tagggtttcg
    10261 ctcatgtgtt gagcatataa gaaaccctta gtatgtattt gtatttgtaa aatacttcta
    10321 tcaataaaat ttctaattcc taaaaccaaa atccagtact aaaatccaga tctcctaaag
    10381 tccctataga tctttgtcgt gaatataaac cagacacgag acgactaaac ctggagccca
    10441 gacgccgttc gaagctagaa gtaccgctta ggcaggaggc cgttagggaa aagatgctaa
    10501 ggcagggttg gttacgttga ctcccccgta ggtttggttt aaatatgatg aagtggacgg
    10561 aaggaaggag gaagacaagg aaggataagg ttgcaggccc tgtgcaaggt aagaagatgg
    10621 aaatttgata gaggtacgct actatactta tactatacgc taagggaatg cttgtattta
    10681 taccctatac cccctaataa ccccttatca atttaagaaa taatccgcat aagcccccgc
    10741 ttaaaaattg gtatcagagc catgaatagg tctatgacca aaactcaaga ggataaaacc
    10801 tcaccaaaat acgaaagagt tcttaactct aaagataaaa gatctttcaa gatcaaaact
    10861 agttccctca caccggtgac ggggatcgca tgcgattcga gatctagctt ggcgtaatca
    10921 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
    10981 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
    11041 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
    11101 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
    11161 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
    11221 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
    11281 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
    11341 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
    11401 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
    11461 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
    11521 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
    11581 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
    11641 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
    11701 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
    11761 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
    11821 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
    11881 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
    11941 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
    12001 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
    12061 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
    12121 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
    12181 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
    12241 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
    12301 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
    12361 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
    12421 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
    12481 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
    12541 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
    12601 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
    12661 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
    12721 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
    12781 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
    12841 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
    12901 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
    12961 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
    13021 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgccagt
    13081 gccaagttaa ttcg
//
