LOCUS       sequence-238953-       13442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway destination vector encoding Maize ubiquitin promoter driving
            Cas9 and 35S promoter driving hygromycin for selection in plants..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13442)
  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 13442)
  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: pMH-Cas9-gate
FEATURES             Location/Qualifiers
     source          1..13442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(126..194,195..986)
                     /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(344..363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1157..1745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1646..1665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1899..1916
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2033..2054
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2069..2074,2075..2092,2093..2099)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2107..2123
                     /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)."
     primer_bind     2112..2134
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2131..2147
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2131..2147
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2701..2877
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(2947..3972)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(2947..3972)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(4002..4347)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(4039..4059)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4380..4632)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     4659..4675
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4660..4676
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4753..4877
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(4902..4907,4908..4925,4926..4932)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             4986..5645
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5052..5071)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             5987..6292
                     /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     6171..6190
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(6333..6457)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        6480..8472
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     protein_bind    8495..8519
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             8546..8566
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8567..12670
                     /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             8567..12667
                     /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(12687..12711)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      12724..12733
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        13131..13310
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     primer_bind     complement(13378..13394)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13378..13394)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
       61 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
      961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1801 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     1861 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     1921 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1981 gctggcacga caggtttccc gactggaaag cgggcagtga gcacaacgca attaatgtga
     2041 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     2101 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     2161 agctagatct cgagatatcg catgcgatcc ccgtcaccgg tgtgagggaa ctagttttga
     2221 tcttgaaaga tcttttatct ttagagttaa gaactctttc gtattttggt gaggttttat
     2281 cctcttgagt tttggtcata gacctattca tggctctgat accaattttt aagcgggggc
     2341 ttatgcggat tatttcttaa attgataagg ggttattagg gggtataggg tataaataca
     2401 agcattccct tagcgtatag tataagtata gtagcgtacc tctatcaaat ttccatcttc
     2461 ttaccttgca cagggcctgc aaccttatcc ttccttgtct tcctccttcc ttccgtccac
     2521 ttcatcatat ttaaaccaaa cctacggggg agtcaacgta accaaccctg ccttagcatc
     2581 ttttccctaa cggcctcctg cctaagcggt acttctagct tcgaacggcg tctgggctcc
     2641 aggtttagtc gtctcgtgtc tggtttatat tcacgacaaa gatctatagg gactttagga
     2701 gatctggatt ttagtactgg attttggttt taggaattag aaattttatt gatagaagta
     2761 ttttacaaat acaaatacat actaagggtt tcttatatgc tcaacacatg agcgaaaccc
     2821 tataagaacc ctaatttccc ttatcgggaa actactcaca cattatttat ggagaaaata
     2881 gagagagata gatttgtaga gagagactgg tgatttcagc gtaccgtccg aaccccagag
     2941 tcccgctcat tcctttgccc tcggacgagt gctggggcgt cggtttccac tatcggcgag
     3001 tacttctaca cagccatcgg tccagacggc cgcgcttctg cgggcgattt gtgtacgccc
     3061 gacagtcccg gctccggatc ggacgattgc gtcgcatcga ccctgcgccc aagctgcatc
     3121 atcgaaattg ccgtcaacca agctctgata gagttggtca agaccaatgc ggagcatata
     3181 cgcccggagc cgcggcgatc ctgcaagctc cggatgcctc cgctcgaagt agcgcgtctg
     3241 ctgctccata caagccaacc acggcctcca gaagaagatg ttggcgacct cgtattggga
     3301 atccccgaac atcgcctcgc tccagtcaat gaccgctgtt atgcggccat tgtccgtcag
     3361 gacattgttg gagccgaaat ccgcgtgcac gaggtgccgg acttcggggc agtcctcggc
     3421 ccaaagcatc agctcatcga gagcctgcgc gacggacgca ctgacggtgt cgtccatcac
     3481 agtttgccag tgatacacat ggggatcagc aatcgcgcat atgaaatcac gccatgtagt
     3541 gtattgaccg attccttgcg gtccgaatgg gccgaacccg ctcgtctggc taagatcggc
     3601 cgcagcgatc gcatccatgg cctccgcgac cggctgcaga acagcgggca gttcggtttc
     3661 aggcaggtct tgcaacgtga caccctgtgc acggcgggag atgcaatagg tcaggctctc
     3721 gctgaattcc ccaatgtcaa gcacttccgg aatcgggagc gcggccgatg caaagtgccg
     3781 ataaacataa cgatctttgt agaaaccatc ggcgcagcta tttacccgca ggacatatcc
     3841 acgccctcct acatcgaagc tgaaagcacg agattcttcg ccctccgaga gctgcatcag
     3901 gtcggagacg ctgtcgaact tttcgatcag aaacttctcg acagacgtcg cggtgagttc
     3961 aggctttttc atgcctgcag gtcggttctc catgcttggg ctgtcctctc caaatgaaat
     4021 gaacttcctt atatagagga agggtcttgc gaaggatagt gggattgtgc gtcatccctt
     4081 acgtcagtgg agatgtcaca tcaatccact tgctttgaag acgtggttgg aacgtcttct
     4141 ttttccacga tgctcctcgt gggtgggggt ccatctttgg gaccactgtc ggcagaggca
     4201 tcttgaatga tagcctttcc tttatcgcaa tgatggcatt tgtaggagcc accttccttt
     4261 tctactgtcc tttcgatgaa gtgacagata gctgggcaat ggaatccgag gaggtttccc
     4321 gaaattaccc tttgttgaaa agtctcaata gccctttggt cttctgagac tgtatccccg
     4381 atctagtaac atagatgaca ccgcgcgcga taatttatcc tagtttgcgc gctatatttt
     4441 gttttctatc gcgtattaaa tgtataattg cgggactcta atcataaaaa cccatctcat
     4501 aaataacgtc atgcattaca tgttaattat tacatgctta acgtaattca acagaaatta
     4561 tatgataatc atcgcaagac cggcaacagg attcaatctt aagaaacttt attgccaaat
     4621 gtttgaacga tcttggagta ggggtaccgg gccccccctc gaggtcgacg gtatcgataa
     4681 gcttgatatc gaattcctgc agccaaactg aaggcgggaa acgacaatct gatccaagct
     4741 caagctaagc ttacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat
     4801 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
     4861 catatccagt cactatggcg gccgcattag gcaccccagg ctttacactt tatgcttccg
     4921 gctcgtataa tgtgtggatt ttgagttagg atccgtcgag attttcagga gctaaggaag
     4981 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     5041 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     5101 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     5161 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag
     5221 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     5281 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     5341 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     5401 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     5461 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     5521 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     5581 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     5641 cgtaaacgcg tggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg
     5701 atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggta
     5761 tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc
     5821 atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt
     5881 ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt
     5941 attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc agtttaaggt
     6001 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
     6061 tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
     6121 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
     6181 caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca
     6241 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg
     6301 ctcccttata cacagccagt ctgcaggtcg accatagtga ctggatatgt tgtgttttac
     6361 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat
     6421 tttacgtttc tcgttcagct ttcttgtaca aagtggttct agagatcaag cttgcatgcc
     6481 tgcagtgcag cgtgacccgg tcgtgcccct ctctagagat aatgagcatt gcatgtctaa
     6541 gttataaaaa attaccacat attttttttg tcacacttgt ttgaagtgca gtttatctat
     6601 ctttatacat atatttaaac tttactctac gaataatata atctatagta ctacaataat
     6661 atcagtgttt tagagaatca tataaatgaa cagttagaca tggtctaaag gacaattgag
     6721 tattttgaca acaggactct acagttttat ctttttagtg tgcatgtgtt ctcctttttt
     6781 tttgcaaata gcttcaccta tataatactt catccatttt attagtacat ccatttaggg
     6841 tttagggtta atggttttta tagactaatt tttttagtac atctatttta ttctatttta
     6901 gcctctaaat taagaaaact aaaactctat tttagttttt ttatttaata atttagatat
     6961 aaaatagaat aaaataaagt gactaaaaat taaacaaata ccctttaaga aattaaaaaa
     7021 actaaggaaa catttttctt gtttcgagta gataatgcca gcctgttaaa cgccgtcgac
     7081 gagtctaacg gacaccaacc agcgaaccag cagcgtcgcg tcgggccaag cgaagcagac
     7141 ggcacggcat ctctgtcgct gcctctggac ccctctcgag agttccgctc caccgttgga
     7201 cttgctccgc tgtcggcatc cagaaattgc gtggcggagc ggcagacgtg agccggcacg
     7261 gcaggcggcc tcctcctcct ctcacggcac cggcagctac gggggattcc tttcccaccg
     7321 ctccttcgct ttcccttcct cgcccgccgt aataaataga caccccctcc acaccctctt
     7381 tccccaacct cgtgttgttc ggagcgcaca cacacacaac cagatctccc ccaaatccac
     7441 ccgtcggcac ctccgcttca aggtacgccg ctcgtcctcc cccccccccc ctctctacct
     7501 tctctagatc ggcgttccgg tccatggtta gggcccggta gttctacttc tgttcatgtt
     7561 tgtgttagat ccgtgtttgt gttagatccg tgctgctagc gttcgtacac ggatgcgacc
     7621 tgtacgtcag acacgttctg attgctaact tgccagtgtt tctctttggg gaatcctggg
     7681 atggctctag ccgttccgca gacgggatcg atttcatgat tttttttgtt tcgttgcata
     7741 gggtttggtt tgcccttttc ctttatttca atatatgccg tgcacttgtt tgtcgggtca
     7801 tcttttcatg cttttttttg tcttggttgt gatgatgtgg tctggttggg cggtcgttct
     7861 agatcggagt agaattctgt ttcaaactac ctggtggatt tattaatttt ggatctgtat
     7921 gtgtgtgcca tacatattca tagttacgaa ttgaagatga tggatggaaa tatcgatcta
     7981 ggataggtat acatgttgat gcgggtttta ctgatgcata tacagagatg ctttttgttc
     8041 gcttggttgt gatgatgtgg tgtggttggg cggtcgttca ttcgttctag atcggagtag
     8101 aatactgttt caaactacct ggtgtattta ttaattttgg aactgtatgt gtgtgtcata
     8161 catcttcata gttacgagtt taagatggat ggaaatatcg atctaggata ggtatacatg
     8221 ttgatgtggg ttttactgat gcatatacat gatggcatat gcagcatcta ttcatatgct
     8281 ctaaccttga gtacctatct attataataa acaagtatgt tttataatta ttttgatctt
     8341 gatatacttg gatgatggca tatgcagcag ctatatgtgg atttttttag ccctgccttc
     8401 atacgctatt tatttgcttg gtactgtttc ttttgtcgat gctcaccctg ttgtttggtg
     8461 ttacttctgc aggtcgactc tagaggatcc cggggcaagt ttgtacaaaa aagcaggctc
     8521 cgcggccgcc cccttcacca tggccccaaa gaagaagcgc aaggtcgaca agaagtactc
     8581 catcggcctc gacatcggca ccaattctgt tggctgggcc gtgatcaccg acgagtacaa
     8641 ggtgccgtcc aagaagttca aggtcctcgg caacaccgac cgccactcca tcaagaagaa
     8701 tctcatcggc gccctgctgt tcgactctgg cgagacagcc gaggctacaa ggctcaagag
     8761 gaccgctaga cgcaggtaca ccaggcgcaa gaaccgcatc tgctacctcc aagagatctt
     8821 ctccaacgag atggccaagg tggacgacag cttcttccac aggctcgagg agagcttcct
     8881 cgtcgaggag gacaagaagc acgagcgcca tccgatcttc ggcaacatcg tggatgaggt
     8941 ggcctaccac gagaagtacc cgaccatcta ccacctccgc aagaagctcg tcgactccac
     9001 cgataaggcc gacctcaggc tcatctacct cgccctcgcc cacatgatca agttcagggg
     9061 ccacttcctc atcgagggcg acctcaaccc ggacaactcc gatgtggaca agctgttcat
     9121 ccagctcgtg cagacctaca accagctgtt cgaggagaac ccgatcaacg cctctggcgt
     9181 tgacgccaag gctattctct ctgccaggct ctctaagtcc cgcaggctcg agaatctgat
     9241 cgcccaactt ccgggcgaga agaagaatgg cctcttcggc aacctgatcg ccctctctct
     9301 tggcctcacc ccgaacttca agtccaactt cgacctcgcc gaggacgcca agctccagct
     9361 ttccaaggac acctacgacg acgacctcga caatctcctc gcccagattg gcgatcagta
     9421 cgccgatctg ttcctcgccg ccaagaatct ctccgacgcc atcctcctca gcgacatcct
     9481 cagggtgaac accgagatca ccaaggcccc actctccgcc tccatgatca agaggtacga
     9541 cgagcaccac caggacctca cactcctcaa ggccctcgtg agacagcagc tcccagagaa
     9601 gtacaaggag atcttcttcg accagtccaa gaacggctac gccggctaca tcgatggcgg
     9661 cgcttctcaa gaggagttct acaagttcat caagccgatc ctcgagaaga tggacggcac
     9721 cgaggagctg ctcgtgaagc tcaatagaga ggacctcctc cgcaagcagc gcaccttcga
     9781 taatggctcc atcccgcacc agatccacct cggcgagctt catgctatcc tccgcaggca
     9841 agaggacttc tacccgttcc tcaaggacaa ccgcgagaag attgagaaga tcctcacctt
     9901 ccgcatcccg tactacgtgg gcccgctcgc caggggcaac tccaggttcg cctggatgac
     9961 cagaaagtcc gaggagacaa tcaccccctg gaacttcgag gaggtggtgg ataagggcgc
    10021 ctctgcccag tctttcatcg agcgcatgac caacttcgac aagaacctcc cgaacgagaa
    10081 ggtgctcccg aagcactcac tcctctacga gtacttcacc gtgtacaacg agctgaccaa
    10141 ggtgaagtac gtgaccgagg ggatgaggaa gccagctttc cttagcggcg agcaaaagaa
    10201 ggccatcgtc gacctgctgt tcaagaccaa ccgcaaggtg accgtgaagc agctcaagga
    10261 ggactacttc aagaaaatcg agtgcttcga ctccgtcgag atctccggcg tcgaggatag
    10321 gttcaatgcc tccctcggga cctaccacga cctcctcaag attatcaagg acaaggactt
    10381 cctcgacaac gaggagaacg aggacatcct cgaggacatc gtgctcaccc tcaccctctt
    10441 cgaggaccgc gagatgatcg aggagcgcct caagacatac gcccacctct tcgacgacaa
    10501 ggtgatgaag cagctgaagc gcaggcgcta taccggctgg ggcaggctct ctaggaagct
    10561 catcaacggc atccgcgaca agcagtccgg caagacgatc ctcgacttcc tcaagtccga
    10621 cggcttcgcc aaccgcaact tcatgcagct catccacgac gactccctca ccttcaagga
    10681 ggacatccaa aaggcccagg tgtccggcca aggcgattcc ctccatgaac atatcgccaa
    10741 tctcgccggc tccccggcta tcaagaaggg cattctccag accgtgaagg tggtggacga
    10801 gctggtgaag gtgatgggca ggcacaagcc agagaacatc gtgatcgaga tggcccgcga
    10861 gaaccagacc acacagaagg gccaaaagaa ctcccgcgag cgcatgaaga ggatcgagga
    10921 gggcattaag gagctgggct cccagatcct caaggagcac ccagtcgaga acacccagct
    10981 ccagaacgag aagctctacc tctactacct ccagaacggc cgcgacatgt acgtggacca
    11041 agagctggac atcaaccgcc tctccgacta cgacgtggac catattgtgc cgcagtcctt
    11101 cctgaaggac gactccatcg acaacaaggt gctcacccgc tccgacaaga acaggggcaa
    11161 gtccgataac gtgccgtccg aagaggtcgt caagaagatg aagaactact ggcgccagct
    11221 cctcaacgcc aagctcatca cccagaggaa gttcgacaac ctcaccaagg ccgagagagg
    11281 cggcctttcc gagcttgata aggccggctt catcaagcgc cagctcgtcg agacacgcca
    11341 gatcacaaag cacgtggccc agatcctcga ctcccgcatg aacaccaagt acgacgagaa
    11401 cgacaagctc atccgcgagg tgaaggtcat caccctcaag tccaagctcg tgtccgactt
    11461 ccgcaaggac ttccagttct acaaggtgcg cgagatcaac aactaccacc acgcccacga
    11521 cgcctacctc aatgccgtgg tgggcacagc cctcatcaag aagtacccaa agctcgagtc
    11581 cgagttcgtg tacggcgact acaaggtgta cgacgtgcgc aagatgatcg ccaagtccga
    11641 gcaagagatc ggcaaggcga ccgccaagta cttcttctac tccaacatca tgaatttctt
    11701 caagaccgag atcacgctcg ccaacggcga gattaggaag aggccgctca tcgagacaaa
    11761 cggcgagaca ggcgagatcg tgtgggacaa gggcagggat ttcgccacag tgcgcaaggt
    11821 gctctccatg ccgcaagtga acatcgtgaa gaagaccgag gttcagaccg gcggcttctc
    11881 caaggagtcc atcctcccaa agcgcaactc cgacaagctg atcgcccgca agaaggactg
    11941 ggacccgaag aagtatggcg gcttcgattc tccgaccgtg gcctactctg tgctcgtggt
    12001 tgccaaggtc gagaagggca agagcaagaa gctcaagtcc gtcaaggagc tgctgggcat
    12061 cacgatcatg gagcgcagca gcttcgagaa gaacccaatc gacttcctcg aggccaaggg
    12121 ctacaaggag gtgaagaagg acctcatcat caagctcccg aagtacagcc tcttcgagct
    12181 tgagaacggc cgcaagagaa tgctcgcctc tgctggcgag cttcagaagg gcaacgagct
    12241 tgctctcccg tccaagtacg tgaacttcct ctacctcgcc tcccactacg agaagctcaa
    12301 gggctcccca gaggacaacg agcaaaagca gctgttcgtc gagcagcaca agcactacct
    12361 cgacgagatc atcgagcaga tctccgagtt ctccaagcgc gtgatcctcg ccgatgccaa
    12421 cctcgataag gtgctcagcg cctacaacaa gcaccgcgat aagccaattc gcgagcaggc
    12481 cgagaacatc atccacctct tcaccctcac caacctcggc gctccagccg ccttcaagta
    12541 cttcgacacc accatcgacc gcaagcgcta cacctctacc aaggaggttc tcgacgccac
    12601 cctcatccac cagtctatca caggcctcta cgagacacgc atcgacctct cacaactcgg
    12661 cggcgattga aagggtgggc gcgccgaccc agctttcttg tacaaagtgg tgatatcccg
    12721 cggccatggc ggccgggagc atgcgacgtc gatctaactg actagccgcg gccatgctag
    12781 agtccgcaaa aatcaccagt ctctctctac aaatctatct ctctctattt ttctccagaa
    12841 taatgtgtga gtagttccca gataagggaa ttagggttct tatagggttt cgctcatgtg
    12901 ttgagcatat aagaaaccct tagtatgtat ttgtatttgt aaaatacttc tatcaataaa
    12961 atttctaatt cctaaaacca aaatccagtg acctgcaggc atgcgacgtc gggccctcta
    13021 gaggatcccc gggtaccgcg aattatcgat catgagcgga gaattaaggg agtcacgtta
    13081 tgacccccgc cgatgacgcg ggacaagccg ttttacgttt ggaactgaca gaaccgcaac
    13141 gttgaaggag ccactgagcc gcgggtttct ggagtttaat gagctaagca catacgtcag
    13201 aaaccattat tgcgcgttca aaagtcgcct aaggtcacta tcagctagca aatatttctt
    13261 gtcaaaaatg ctccactgac gttccataaa ttcccctcgg tatccaatta gagtctcata
    13321 ttcactctca actcgaatcc cccctatgaa aaagcctgaa ctcaccgcga cgtgggggat
    13381 ccactagttc tagagcggcc gccaccgcgg tggagctcga attagcttgg cactggcagg
    13441 tg
//
