| 质粒名称: | pFGC-pcoCas9 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 52256 |
| 存储实验室: | Jen Sheen |
|
载体骨架基本信息 |
|
| 载体名称: | pFGC-RCS |
| 完整质粒大小: | 13330bp |
| 载体类型: | CRISPR ; Plant expression |
| 筛选标记: | Basta |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | 35SPPDK:pcoCas9:NOS cassette/Cas9/plant codon optimized Cas9 |
| 物种来源: | Synthetic |
| 启动子: | constitutive 35SPPDK promoter |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | StuI(未破坏) |
| 3’克隆位点: | StuI (未破坏) |
| 5’测序引物 | 5’ AATAAAAACTGACTCGGA 3’ |
| 3’测序引物 | 5’ AGTTGGGTAACGCCAGGG 3’ |
|
文献引用方法 |
|
| 材料和方法部分: | pFGC-pcoCas9 was a gift from Jen Sheen (Addgene plasmid # 52256 ; http://n2t.net/addgene:52256 ; RRID:Addgene_52256) |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pFGC-pcoCas9 13330 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION A binary plasmid containing 35SPPDK:pcoCas9:NOS cassette for plant
expression of Cas9 and multiple cloning sites for insertion of guide
RNA.
ACCESSION .
VERSION .
KEYWORDS pFGC-pcoCas9
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13330)
TITLE Sheen lab CRISPR plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 13330)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..13330
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 235..487
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
CDS complement(496..543)
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS complement(join(544..3207,3397..4833))
/codon_start=1
/product="Cas9 endonuclease from the Streptococcus pyogenes
Type II CRISPR/Cas system"
/label=pcoCas9
/note="plant codon-optimized Cas9 gene containing the
potato IV2 intron"
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
intron 3208..3396
/label=IV2 intron
/note="modified second intron of the potato ST-LS1 gene
(Vancanneyt et al., 1990)"
CDS complement(4858..4878)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS complement(4885..4932)
/codon_start=1
/product="two tandem FLAG(R) epitope tags"
/label=2xFLAG
/translation="DYKDDDDKDYKDDDDK"
promoter complement(4944..5453)
/label=35SPPDK promoter
/note="hybrid promoter consisting of the cauliflower mosaic
virus 35S enhancer fused to the maize C4PPDK basal promoter
(Yoo et al., 2007)"
primer_bind complement(5516..5533)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(5516..5532)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5525..5547)
/label=M13/pUC Forward
/note="In lacZ gene"
misc_feature 5735..5759
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 7060..7689
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 8118..9191
/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"
rep_origin 9257..9451
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
primer_bind complement(9687..9709)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 9795..9935
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 9850..9869
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(9950..9967)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(10121..10709)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(10201..10220)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(10796..11590)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 12015..12039
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
terminator 12125..12377
/label=MAS terminator
/note="mannopine synthase terminator"
CDS complement(12387..12938)
/codon_start=1
/gene="Streptomyces hygroscopicus bar"
/product="phosphinothricin acetyltransferase"
/label=BlpR
/note="confers resistance to bialophos or phosphinothricin"
/translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
WQLDFSLPVPPRPVLPVTEI"
promoter complement(12944..13324)
/label=MAS promoter
/note="mannopine synthase promoter (Velten et al., 1984)"
ORIGIN
1 gaattcctcg agtacgtagg atccatttaa attctagagg cgcgccgata tcctctctta
61 aggtagcgag ctcttaatta atagggataa cagggtaatt aactataacg gtcctaaggt
121 agcgatggca aacagctatt atgggtatta tgggtggttc tttatgcgga cactgacggc
181 tttatgcctg caggtcgcga gcgatcgcgg taccgcccgg gcgtcgacag gcctgatcta
241 gtaacataga tgacaccgcg cgcgataatt tatcctagtt tgcgcgctat attttgtttt
301 ctatcgcgta ttaaatgtat aattgcggga ctctaatcat aaaaacccat ctcataaata
361 acgtcatgca ttacatgtta attattacat gcttaacgta attcaacaga aattatatga
421 taatcatcgc aagaccggca acaggattca atcttaagaa actttattgc caaatgtttg
481 aacgatctgc agtcacttct tcttcttagc ctgtccagcc ttcttggtag cagctggtct
541 cttatctcct ccaagctgag aaagatcgat tctggtctcg taaagtccgg tgatagactg
601 gtggataagg gtagcatcaa gaacctcctt ggtagaggtg tatctttttc tatcgatggt
661 ggtatcgaag tacttgaaag cagctggagc accaaggttg gtaagggtga aaaggtggat
721 gatgttctca gcctgctctc tgattggctt atctctgtgc ttgttgtaag cagaaagaac
781 cttatcaagg ttagcatcag caaggataac tctcttagag aactcagaga tttgctcgat
841 gatctcatca aggtagtgct tgtgctgctc aacgaaaagc tgcttctgct cgttatcctc
901 tggagatccc ttcaacttct cgtagtgaga agcaaggtaa aggaagttaa cgtacttaga
961 tggaagagca agctcgtttc ccttctgaag ctctccagca gaagcaagca ttctctttct
1021 tccgttctca agctcgaaaa gagagtactt tggcaacttg atgataagat ccttcttaac
1081 ctccttgtat cccttagcct caaggaaatc gattgggttc ttctcgaaag aagaacgctc
1141 catgatggtg attccaagaa gctccttaac agacttcaac ttcttagact ttcccttctc
1201 aaccttagca acaacaagaa cagagtaagc aacggttgga gaatcgaatc ctccgtactt
1261 ctttgggtcc caatccttct ttctagcgat caacttatca gagtttctct ttggaaggat
1321 agactcctta gagaatcctc cggtctgaac ctcggttttc ttaacgatgt taacctgtgg
1381 catagaaaga acctttctaa cggtagcgaa atctcttccc ttatcccaaa cgatctctcc
1441 ggtctctccg ttggtctcga taagtggtct ctttctgatc tctccgttag caagggtgat
1501 ctcggtcttg aagaagttca tgatgttaga gtagaagaag tacttagcgg tagcctttcc
1561 gatctcctgc tcagacttag cgatcatctt tctaacatcg taaaccttgt aatctccgta
1621 aacgaactca gactccaact ttgggtactt cttgataaga gcggttccaa caacagcgtt
1681 aaggtaagca tcgtgagcgt ggtggtagtt gttgatctct ctaaccttgt agaactggaa
1741 atcctttctg aaatcagaaa ccaacttaga cttcaaggtg ataaccttaa cctctctgat
1801 caacttatcg ttctcatcgt acttggtgtt catacgagaa tcaaggatct gagcaacgtg
1861 cttggtgatc tgtctggtct caacaagctg tctcttgatg aatccagcct tatcaagctc
1921 agaaagtcct cctctctcag ccttggtaag gttatcgaac tttctctggg tgatcaactt
1981 agcgttaaga agctgtctcc agtagttctt catcttctta acaacctcct cagatggaac
2041 gttatcagac tttcctctgt tcttatcaga acgggtaaga accttgttat cgatagaatc
2101 atccttcaag aaagactgtg gaacgatgtg atcaacatcg taatcagaaa gtctgttgat
2161 gtcaagctcc tgatcaacgt acatatctct tccgttctga aggtagtaaa ggtacaactt
2221 ctcgttctga agctgggtgt tctcaactgg gtgctccttc aagatttgag atccaagctc
2281 cttgattccc tcctcgattc tcttcattct ctcacgagag ttcttctgtc ccttctgggt
2341 ggtctggttc tctctagcca tctcgataac gatgttctct ggcttgtgtc tacccataac
2401 cttaacaagc tcatcaacaa ccttaacggt ctgaaggatt cccttcttga tagctggaga
2461 tccagcaagg ttagcgatgt gctcgtgaag agaatctccc tgtccagaaa cctgagcctt
2521 ctggatgtcc tccttgaagg taagagaatc atcgtggata agctgcatga agtttctgtt
2581 agcgaatcca tcagacttca agaaatcaag gatggtcttt ccagactgct tatctctgat
2641 tccgttgatc aacttacgag aaagtctacc ccatccggtg tatcttcttc tcttcaactg
2701 cttcataacc ttatcatcga aaaggtgagc gtaggtcttg agtctctcct cgatcatctc
2761 tctatcctcg aaaagggtaa gggtaagaac gatgtcctca aggatgtcct cgttctcctc
2821 gttatcaagg aaatccttat ccttgatgat cttcaaaaga tcgtggtagg ttccaagaga
2881 agcgttgaat ctatcctcaa ctccagagat ttcaacagaa tcgaagcact cgatcttctt
2941 gaagtaatcc tccttcaact gcttaacggt aacctttctg ttggtcttga aaagaagatc
3001 aacgatagcc ttcttctgct ctccagaaag gaaagctggc tttctcattc cctcggtaac
3061 gtacttaacc ttggtaagct cgttgtaaac ggtgaagtac tcgtaaagaa gagagtgctt
3121 tggaagaacc ttctcgtttg gaaggttctt atcgaagttg gtcattctct cgatgaaaga
3181 ctgagcagaa gctcccttat caacaacctg cacatcaaca aattttggtc atatattaga
3241 aaagttataa attaaaatat acacacttat aaactacaga aaagcaattg ctatatacta
3301 cattctttta ttttgaaaaa aatatttgaa atattatatt actactaatt aatgataatt
3361 attatatata tatcaaaggt agaagcagaa acttacctcc tcgaagttcc aaggggtgat
3421 ggtctcctca gactttctgg tcatccaagc gaaacgagag tttcctctag caagtggtcc
3481 aacgtagtat gggattctga aggtaaggat cttctcgatc ttctctctgt tatccttcaa
3541 gaatgggtag aaatcctcct gtctacgaag gatagcgtga agctctccaa ggtggatctg
3601 gtgtgggata gatccgttat cgaaggttct ctgctttcta agaagatcct ctctgttcaa
3661 cttaacaaga agctcctcgg ttccatccat cttctcaagg attggcttga tgaacttgta
3721 gaactcctcc tgagaagctc ctccatcgat gtatccagcg tatccgttct tagactgatc
3781 gaagaagatt tccttgtact tctctggaag ctgctgtcta acaagagcct tcaaaagggt
3841 aagatcctgg tggtgctcat cgtatctctt gatcatagaa gcagaaagtg gagccttggt
3901 gatctcggtg ttaactctaa ggatgtcaga aagaaggata gcatcagaaa ggttcttagc
3961 agcaaggaaa agatcagcgt actgatctcc gatctgagca agaaggttat caagatcatc
4021 atcgtaggta tccttagaaa gctgcaactt agcatcctca gcaagatcga agttagactt
4081 gaagtttggg gtaagtccaa gagaaagagc gataaggttt ccgaaaagtc cgttcttctt
4141 ctctcctgga agctgagcga taaggttctc aagtctacga gacttagaaa gtctagcaga
4201 aaggatagcc ttagcatcaa ctccagaagc gttgattggg ttctcctcga aaagctggtt
4261 gtaggtctga acaagctgga tgaacaactt atcaacatca gagttatctg ggttaaggtc
4321 tccctcgata aggaagtgtc ctctgaactt gatcatgtga gcaagagcaa ggtagataag
4381 tctaagatca gccttatcgg tagaatcaac caacttcttt ctaaggtggt agatggttgg
4441 gtacttctcg tggtaagcaa cctcatcaac gatgtttccg aagattgggt gtctctcgtg
4501 cttcttatcc tcctcaacaa ggaaagactc ctcaagtctg tggaagaaag aatcatcaac
4561 cttagccatc tcgttagaga agatttcctg aaggtagcag attctgttct ttcttctggt
4621 gtatcttctt ctagcggttc tcttcaatct ggtagcctca gcggtctctc cagaatcgaa
4681 aagaagagca ccgataaggt tcttcttgat agagtgtcta tcggtgtttc caagaacctt
4741 gaacttctta gatggaacct tgtactcatc ggtgataaca gcccatccaa cagagttggt
4801 tccgatgtca agtccgatag agtacttctt atcagcagct ggaactccgt ggattccaac
4861 ctttctcttc ttctttggag ccatcttatc atcatcatcc ttgtaatcct tatcatcatc
4921 atccttgtaa tccatggatc cacggagcaa ggggagaggg agccgagagc tgcttactac
4981 gtgaccgccc cgcgcctgct gctgtccgga gctccaacgg agctggagag ggggtgtgtg
5041 tggcgagaga gtacagcgaa aaggtgagag gcgaacgggt gaatcctgca cgcggaatgc
5101 ctttattctt ataccatccg tggggcgcga cccaagcttg ggcccaagct tggggcgaag
5161 gatagtggga ttgtgcgtca tcccttacgt cagtggagat atcacatcaa tccacttgct
5221 ttgaagacgt ggttggaacg tcttcttttt ccacgatgct ccttgtgggt gggggtccat
5281 ctttgggacc actgtcggca gaggcatctt gaacgatagc ctttccttta tcgcaatgat
5341 ggcatttgta ggtgccacct tccttttcta ctgtcctttt gatgaagtga cagatagctg
5401 ggcaatggaa tccgaggagg tttcccgata ttaccctttg ttgaaaagtc tcaatagccc
5461 tttggtcttc tgagactgta tctttgatat tcttggagta aggcctaagc ttggcactgg
5521 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
5581 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
5641 cccaacagtt gcgcagcctg aatggcgaat gctagagcag cttgagcttg gatcagattg
5701 tcgtttcccg ccttcagttt aaactatcag tgtttgacag gatatattgg cgggtaaacc
5761 taagagaaaa gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat
5821 ccgttcgtcc atttgtatgt gcatgccaac cacagggttc ccctcgggat caaagtactt
5881 tgatccaacc cctccgctgc tatagtgcag tcggcttctg acgttcagtg cagccgtctt
5941 ctgaaaacga catgtcgcac aagtcctaag ttacgcgaca ggctgccgcc ctgccctttt
6001 cctggcgttt tcttgtcgcg tgttttagtc gcataaagta gaatacttgc gactagaacc
6061 ggagacatta cgccatgaac aagagcgccg ccgctggcct gctgggctat gcccgcgtca
6121 gcaccgacga ccaggacttg accaaccaac gggccgaact gcacgcggcc ggctgcacca
6181 agctgttttc cgagaagatc accggcacca ggcgcgaccg cccggagctg gccaggatgc
6241 ttgaccacct acgccctggc gacgttgtga cagtgaccag gctagaccgc ctggcccgca
6301 gcacccgcga cctactggac attgccgagc gcatccagga ggccggcgcg ggcctgcgta
6361 gcctggcaga gccgtgggcc gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt
6421 tcgccggcat tgccgagttc gagcgttccc taatcatcga ccgcacccgg agcgggcgcg
6481 aggccgccaa ggcccgaggc gtgaagtttg gcccccgccc taccctcacc ccggcacaga
6541 tcgcgcacgc ccgcgagctg atcgaccagg aaggccgcac cgtgaaagag gcggctgcac
6601 tgcttggcgt gcatcgctcg accctgtacc gcgcacttga gcgcagcgag gaagtgacgc
6661 ccaccgaggc caggcggcgc ggtgccttcc gtgaggacgc attgaccgag gccgacgccc
6721 tggcggccgc cgagaatgaa cgccaagagg aacaagcatg aaaccgcacc aggacggcca
6781 ggacgaaccg tttttcatta ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt
6841 gttcgagccg cccgcgcacg tctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc
6901 tgatgccaag ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg
6961 tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat
7021 gatgcgatga gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt
7081 aaccagaaag gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa
7141 ctcgccgggg ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg
7201 gcggccgtgc gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac
7261 cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg
7321 gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca
7381 agcccttacg acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag
7441 gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc
7501 atcggcggtg aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt
7561 atcacgcagc gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca
7621 gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa
7681 ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg
7741 gccgtccgag cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag
7801 ccatgaagcg ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg
7861 cggtacgcca aggcaagacc attaccgagc tgctatctga atacatcgcg cagctaccag
7921 agtaaatgag caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg
7981 gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc
8041 ggttggccag gcgtaagcgg ctgggttgcc tgccggccct gcaatggcac tggaaccccc
8101 aagcccgagg aatcggcgtg agcggtcgca aaccatccgg cccggtacaa atcggcgcgg
8161 cgctgggtga tgacctggtg gagaagttga aggccgcgca ggccgcccag cggcaacgca
8221 tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag
8281 aatcccggca accgccggca gccggtgcgc cgtcgattag gaagccgccc aagggcgacg
8341 agcaaccaga ttttttcgtt ccgatgctct atgacgtggg cacccgcgat agtcgcagca
8401 tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg acgagctggc gaggtgatcc
8461 gctacgagct tccagacggg cacgtagagg tttccgcagg gccggccggc atggccagtg
8521 tgtgggatta cgacctggta ctgatggcgg tttcccatct aaccgaatcc atgaaccgat
8581 accgggaagg gaagggagac aagcccggcc gcgtgttccg tccacacgtt gcggacgtac
8641 tcaagttctg ccggcgagcc gatggcggaa agcagaaaga cgacctggta gaaacctgca
8701 ttcggttaaa caccacgcac gttgccatgc agcgtacgaa gaaggccaag aacggccgcc
8761 tggtgacggt atccgagggt gaagccttga ttagccgcta caagatcgta aagagcgaaa
8821 ccgggcggcc ggagtacatc gagatcgagc tagctgattg gatgtaccgc gagatcacag
8881 aaggcaagaa cccggacgtg ctgacggttc accccgatta ctttttgatc gatcccggca
8941 tcggccgttt tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa gccagatggt
9001 tgttcaagac gatctacgaa cgcagtggca gcgccggaga gttcaagaag ttctgtttca
9061 ccgtgcgcaa gctgatcggg tcaaatgacc tgccggagta cgatttgaag gaggaggcgg
9121 ggcaggctgg cccgatccta gtcatgcgct accgcaacct gatcgagggc gaagcatccg
9181 ccggttccta atgtacggag cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc
9241 gaaaaggtct ctttcctgtg gatagcacgt acattgggaa cccaaagccg tacattggga
9301 accggaaccc gtacattggg aacccaaagc cgtacattgg gaaccggtca cacatgtaag
9361 tgactgatat aaaagagaaa aaaggcgatt tttccgccta aaactcttta aaacttatta
9421 aaactcttaa aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca gccgaagagc
9481 tgcaaaaagc gcctaccctt cggtcgctgc gctccctacg ccccgccgct tcgcgtcggc
9541 ctatcgcggc cgctggccgc tcaaaaatgg ctggcctacg gccaggcaat ctaccagggc
9601 gcggacaagc cgcgccgtcg ccactcgacc gccggcgccc acatcaaggc accctgcctc
9661 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
9721 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
9781 ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc
9841 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
9901 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
9961 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
10021 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
10081 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
10141 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
10201 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
10261 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
10321 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
10381 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
10441 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
10501 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
10561 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
10621 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
10681 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
10741 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgcattct aggtactaaa
10801 acaattcatc cagtaaaata taatatttta ttttctccca atcaggcttg atccccagta
10861 agtcaaaaaa tagctcgaca tactgttctt ccccgatatc ctccctgatc gaccggacgc
10921 agaaggcaat gtcataccac ttgtccgccc tgccgcttct cccaagatca ataaagccac
10981 ttactttgcc atctttcaca aagatgttgc tgtctcccag gtcgccgtgg gaaaagacaa
11041 gttcctcttc gggcttttcc gtctttaaaa aatcatacag ctcgcgcgga tctttaaatg
11101 gagtgtcttc ttcccagttt tcgcaatcca catcggccag atcgttattc agtaagtaat
11161 ccaattcggc taagcggctg tctaagctat tcgtataggg acaatccgat atgtcgatgg
11221 agtgaaagag cctgatgcac tccgcataca gctcgataat cttttcaggg ctttgttcat
11281 cttcatactc ttccgagcaa aggacgccat cggcctcact catgagcaga ttgctccagc
11341 catcatgccg ttcaaagtgc aggacctttg gaacaggcag ctttccttcc agccatagca
11401 tcatgtcctt ttcccgttcc acatcatagg tggtcccttt ataccggctg tccgtcattt
11461 ttaaatatag gttttcattt tctcccacca gcttatatac cttagcagga gacattcctt
11521 ccgtatcttt tacgcagcgg tatttttcga tcagtttttt caattccggt gatattctca
11581 ttttagccat ttattatttc cttcctcttt tctacagtat ttaaagatac cccaagaagc
11641 taattataac aagacgaact ccaattcact gttccttgca ttctaaaacc ttaaatacca
11701 gaaaacagct ttttcaaagt tgttttcaaa gttggcgtat aacatagtat cgacggagcc
11761 gattttgaaa ccgcggtgat cacaggcagc aacgctctgt catcgttaca atcaacatgc
11821 taccctccgc gagatcatcc gtgtttcaaa cccggcagct tagttgccgt tcttccgaat
11881 agcatcggta acatgagcaa agtctgccgc cttacaacgg ctctcccgct gacgccgtcc
11941 cggactgatg ggctgcctgt atcgagtggt gattttgtgc cgagctgccg gtcggggagc
12001 tgttggctgg ctggtggcag gatatattgt ggtgtaaaca aattgacgct tagacaactt
12061 aataacacat tgcggacgtt tttaatgtac tgaattaacg ccgaattaat tcgagctcgg
12121 atctgataat ttatttgaaa attcataaga aaagcaaacg ttacatgaat tgatgaaaca
12181 atacaaagac agataaagcc acgcacattt aggatattgg ccgagattac tgaatattga
12241 gtaagatcac ggaatttctg acaggagcat gtcttcaatt cagcccaaat ggcagttgaa
12301 atactcaaac cgccccatat gcaggagcgg atcattcatt gtttgtttgg ttgcctttgc
12361 caacatggga gtccaagatt ctgcagtcaa atctcggtga cgggcaggac cggacggggc
12421 ggtaccggca ggctgaagtc cagctgccag aaacccacgt catgccagtt cccgtgcttg
12481 aagccggccg cccgcagcat gccgcggggg gcatatccga gcgcctcgtg catgcgcacg
12541 ctcgggtcgt tgggcagccc gatgacagcg accacgctct tgaagccctg tgcctccagg
12601 gacttcagca ggtgggtgta gagcgtggag cccagtcccg tccgctggtg gcggggggag
12661 acgtacacgg tcgactcggc cgtccagtcg taggcgttgc gtgccttcca ggggcccgcg
12721 taggcgatgc cggcgacctc gccgtccacc tcggcgacga gccagggata gcgctcccgc
12781 agacggacga ggtcgtccgt ccactcctgc ggttcctgcg gctcggtacg gaagttgacc
12841 gtgcttgtct cgatgtagtg gttgacgatg gtgcagaccg ccggcatgtc cgcctcggtg
12901 gcacggcgga tgtcggccgg gcgtcgttct gggctcatcg attcgatttg gtgtatcgag
12961 attggttatg aaattcagat gctagtgtaa tgtattggta atttgggaag atataatagg
13021 aagcaaggct atttatccat ttctgaaaag gcgaaatggc gtcaccgcga gcgtcacgcg
13081 cattccgttc ttgctgtaaa gcgttgtttg gtacactttt gactagcgag gcttggcgtg
13141 tcagcgtatc tattcaaaag tcgttaatgg ctgcggatca agaaaaagtt ggaatagaaa
13201 cagaataccc gcgaaattca ggcccggttg ccatgtccta cacgccgaaa taaacgacca
13261 aattagtaga aaaataaaaa ctgactcgga tacttacgtc acgtcttgcg cactgatttg
13321 aaaaatctca
//
