| 质粒名称: | pPB-C6F/TdTomato |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 140826 |
| 存储实验室: | Hideyuki Okano |
|
载体骨架基本信息 |
|
| 载体名称: | pTRE-Tight-BI |
| 载体出品公司: | Clontech |
| 空载体大小: | -- |
| 完整质粒大小: | 16425 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达 |
| 筛选标记: | 嘌呤霉素 |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称1: | OCT4 |
| 物种来源1: | H. sapiens (human) |
| 插入大小1: | 1080 bp |
| GenBank编号1 | NM_002701.6 |
| 插入基因名称2: | SOX2 |
| 物种来源2: | H. sapiens (human) |
| 插入大小2: | 954 bp |
| GenBank编号2 | NM_003106.4 |
| 插入基因名称3: | KLF4 |
| 物种来源3: | H. sapiens (human) |
| 插入大小3: | 1410 bp |
| GenBank编号3 | NM_004235.6 |
| 插入基因名称4: | C-MYC |
| 物种来源4: | H. sapiens (human) |
| 插入大小4: | 1320 bp |
| GenBank编号4 | NM_002467.6 |
|
文献引用方法 |
|
| 材料和方法部分: | pPB-C6F/TdTomato was a gift from Hideyuki Okano (Addgene plasmid # 140826 ; http://n2t.net/addgene:140826 ; RRID:Addgene_140826) |
| 参考文献部分: | Naive-like ESRRB(+) iPSCs with the Capacity for Rapid Neural Differentiation. Kisa F, Shiozawa S, Oda K, Yoshimatsu S, Nakamura M, Koya I, Kawai K, Suzuki S, Okano H. Stem Cell Reports. 2017 Dec 12;9(6):1825-1838. doi: 10.1016/j.stemcr.2017.10.008. Epub 2017 Nov 9. 10.1016/j.stemcr.2017.10.008 PubMed 29129686 |
买家导航
pPB-C6F/TdTomato载体质粒基本信息
pPB-C6F/TdTomato质粒图谱载体图谱和pPB-C6F/TdTomato载体序列质粒序列多克隆位点信息
pPB-C6F/TdTomato质粒载体简介
pPB-C6F/TdTomato质粒序列载体序列
LOCUS pPB-C6F/TdTomato 16441 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION OCT4, SOX2, KLF4, C-MYC, KLF2, NANOG (C6F) and tdTomato under the
control of TRE (Tet-ON).
ACCESSION .
VERSION .
KEYWORDS pPB-C6F/TdTomato
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 16441)
AUTHORS Kisa F, Shiozawa S, Oda K, Yoshimatsu S, Nakamura M, Koya I, Kawai
K, Suzuki S, Okano H
TITLE Naive-like ESRRB(+) iPSCs with the Capacity for Rapid Neural
Differentiation.
JOURNAL Stem Cell Reports. 2017 Dec 12;9(6):1825-1838. doi:
10.1016/j.stemcr.2017.10.008. Epub 2017 Nov 9.
PUBMED 29129686
REFERENCE 2 (bases 1 to 16441)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..16441
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(140..161)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 352..371
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 600..616
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 623..641
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
repeat_region 660..694
/label=piggyBac left (5') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
primer_bind 1097..1113
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 1120..1138
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 1238..1359
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind 1303..1322
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(1496..2926)
/codon_start=1
/product="tandem dimeric (pseudo-monomeric) derivative of
DsRed (Shaner et al., 2004)"
/label=tdTomato
/note="mammalian codon-optimized"
/translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
RSEGRHHLFLYGMDELYK"
regulatory 2923..2932
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
misc_feature 2946..3498
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
misc_feature 2948..3521
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS complement(4546..4611)
/codon_start=1
/product="2A peptide from foot-and-mouth disease virus
polyprotein"
/label=F2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="VKQTLNFDLLKLAGDVESNPGP"
regulatory 5697..5706
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
promoter 5760..6147
/label=bidirectional TRE promoter
/note="Tet-responsive bidirectional promoter PTight-BI,
consisting of seven tet operator sequences flanked on each
side by the minimal CMV promoter"
protein_bind 5834..5852
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 5870..5888
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 5905..5923
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 5941..5959
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 5977..5995
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 6012..6030
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 6048..6066
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 6164..6184
/label=T3
/note="T3 promoter, forward primer"
promoter 6166..6184
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
CDS 7314..7379
/codon_start=1
/product="2A peptide from foot-and-mouth disease virus
polyprotein"
/label=F2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="VKQTLNFDLLKLAGDVESNPGP"
CDS 8826..8891
/codon_start=1
/product="2A peptide from foot-and-mouth disease virus
polyprotein"
/label=F2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="VKQTLNFDLLKLAGDVESNPGP"
misc_feature 9884..10457
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 10502..11821
/codon_start=1
/product="human c-Myc proto-oncogene"
/label=c-Myc
/translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP
SEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE
LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN
PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS
LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS
PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR
KCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA
TAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA"
misc_feature 11848..12434
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 12435..13034
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(12435..12454)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 12931..12951
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 13176..13409
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(13549..13566)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 13555..13779
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(13826..13842)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 13850..13866
/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(13874..13904)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 13919..13940
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
repeat_region 14167..14229
/label=piggyBac right (3') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
promoter complement(14253..14271)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(14292..14308)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 14316..14332
/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(14340..14370)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 14385..14406
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(14523..14540)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(14694..15282)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(14774..14793)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(15453..16313)
/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 16076..16095
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(16314..16418)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtaccttaac
661 cctagaaaga tagtctgcgt aaaattgacg catgcattct tgaaatattg ctctctcttt
721 ctaaatagcg cgaatccgtc gctgtgcatt taggacatct cagtcgccgc ttggagctcc
781 cgtgaggcgt gcttgtcaat gaggtaagtg tcactgattt tgaactataa cgaccgcgtg
841 agtcaaaatg acgcatgatt atcttttacg tgacttttaa gatttaactc atacgataat
901 tatattgtta tttcatgttc tacttacgtg ataacttatt atatatatat tttcttgtta
961 tagatgtata ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
1021 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
1081 tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc
1141 gaattgggcc ctctagatgc atgctcgacg gtatacagac atgataagat acattgatga
1201 gtttggacaa accacaacta gaatgcactc gacgcgttaa gatacattga tgagtttgga
1261 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
1321 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
1381 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac
1441 aaatgtggta tggctgatta tgatcagtta tctagatgca tgctcgagcg gccgcctact
1501 tgtacagctc gtccatgccg tacaggaaca ggtggtggcg gccctcggag cgctcgtact
1561 gttccacgat ggtgtagtcc tcgttgtggg aggtgatgtc cagcttggtg tccacgtagt
1621 agtagccggg cagttgcacg ggcttcttgg ccatgtagat ggtcttgaac tccaccaggt
1681 agtggccgcc gtccttcagc ttcagggcct ggtggatctc gcccttcagc acgccgtcgc
1741 gggggtacag gcgctcggtg gaggcctccc agcccatggt cttcttctgc attacggggc
1801 cgtcgggggg gaagttggtg ccgcgcatct tcaccttgta gatcagcgtg ccgtcctgca
1861 gggaggagtc ctgggtcacg gtcaccagac cgccgtcctc gaagttcatc acgcgctccc
1921 acttgaagcc ctcggggaag gacagcttct tgtaatcggg gatgtcggcg gggtgcttca
1981 cgtacgcctt ggagccgtac atgaactggg gggacaggat gtcccaggcg aagggcaggg
2041 ggccgccctt ggtcaccttc agcttggcgg tctgggtgcc ctcgtagggg cggccctcgc
2101 cctcgccctc gatctcgaac tcgtggccgt tcatggagcc ctccatgcgc accttgaagc
2161 gcatgaactc tttgatgacg gccatgttgt tgtcctcgga ggaggcggtg ccggagctgc
2221 cgctgccggt gctgccggtg ccatgcccca ggaacaggtg gtggcggccc tcggagcgct
2281 cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt gatgtccagc ttggtgtcca
2341 cgtagtagta gccgggcagt tgcacgggct tcttggccat gtagatggtc ttgaactcca
2401 ccaggtagtg gccgccgtcc ttcagcttca gggcctggtg gatctcgccc ttcagcacgc
2461 cgtcgcgggg gtacaggcgc tcggtggagg cctcccagcc catggtcttc ttctgcatta
2521 cggggccgtc gggggggaag ttggtgccgc gcatcttcac cttgtagatc agcgtgccgt
2581 cctgcaggga ggagtcctgg gtcacggtca ccagaccgcc gtcctcgaag ttcatcacgc
2641 gctcccactt gaagccctcg gggaaggaca gcttcttgta atcggggatg tcggcggggt
2701 gcttcacgta cgccttggag ccgtacatga actgggggga caggatgtcc caggcgaagg
2761 gcagggggcc gcccttggtc accttcagct tggcggtctg ggtgccctcg taggggcggc
2821 cctcgccctc gccctcgatc tcgaactcgt ggccgttcat ggagccctcc atgcgcacct
2881 tgaagcgcat gaactctttg atgacctcct cgcccttgct caccatggtg gcggatccgt
2941 ggccatatta tcatcgtgtt tttcaaagga aaaccacgtc cccgtggttc ggggggccta
3001 gacgtttttt taacctcgac taaacacatg taaagcatgt gtaccgaggc cccagatcag
3061 atcccataca atggggtacc ttctgggcat ccttcagccc cttgttgaat acgcttgagg
3121 agagccattt gactctttcc acaactatcc aactcacaac gtggcactgg ggttgtgccg
3181 cctttgcagg tgtatcttat acacgtggct tttggccgca gaggcacctg tcgccaggtg
3241 gggggttccg ctgcctgcaa agggtcgcta cagacgttgt ttgtcttcaa gaagcttcca
3301 gaggaactgc ttccttcacg acattcaaca gaccttgcat tcctttggcg agaggggaaa
3361 gacccctagg aatgctcgtc aagaagacag ggccaggttt ccgggccctc acattgccaa
3421 aagacggcaa tatggtggaa aataacatat agacaaacgc acaccggcct tattccaagc
3481 ggcttcggcc agtaacgtta gggggggggg agggagaggg gcggactgtg gaaaaaaaag
3541 ggagatccgc tagaagggcg aattcgaatc cggagacgta cgaccggtta acaggcctta
3601 agcgctagat gcatgctcga ctcacacgtc ttcaggttgc atgttcatgg agtagtttag
3661 gaataaatcc atggtttgtg gagtactaaa atacctagtg gtctgctgta ttacattaag
3721 gccttcccca gcagcttcca aagcagcctc caagtcactg gcaggagaat ttggctggaa
3781 ctgcatgcag gactgcagag attcctctcc acagttatag aagggactgt tccaggcctg
3841 attgttccag gattgggtgc accaggtctg agtgttccag gagtggttgc tccaggactg
3901 gatgttctgg gtctggttgc tccaggttga attgttccag gtctggttgc tccacattgg
3961 aaggttccca gtcgggttca ccaggcatcc ctggtggtag gaagagtaga ggctggggta
4021 ggtaggtgct gaggccttct gcgtcacacc attgctattc ttcggccagt tgtttttctg
4081 ccacctctta gatttcattc tctggttctg gaaccaggtc ttcacctgtt tgtagctgag
4141 gttcaggatg ttggagagtt cttgcatctg ctggaggctg aggtatttct gtctctgaaa
4201 tctatcattg agtacacaca gctgggtgga agagaacaca gttctggtct tctgtttctt
4261 gactgggacc ttgtcttcct tttttgcgac actattctct gcagaagtgg gttgtttgcc
4321 tttgggactg gtggaagaat cagggctgtc ctgaataagc agatccatgg aggagggaag
4381 aggagagaca gtctccgtgt gaggcatctc agcagaagac atttgcaagg atggatagtt
4441 ttcttcaggc ccacaaatca caggcatagg tgaagattct ttacagtcgg atgcttcaaa
4501 gcaaggcaag ctttggggac aagctggatc cacactcatg atatccggcc ctgggttgga
4561 ctccacgtct cccgccaact tgagaaggtc aaaattcaaa gtctgtttca ccggtgcccg
4621 cttagcgcga ggatccatgt gccgtttcat gtgcagcgcc aggtgatcgg agcgcgagaa
4681 ggcacgatcg cacagatggc actggaatgg ccggtggccc gtgtgctttc ggtagtggcg
4741 cgtgagctcg tctgagcgcg caaacttcca gccgcagccg tcccagttgc agtggtaggg
4801 cttctcacct gtgtgcgtgc gcagatgcgc cttcagatgc gaactcttgg tgtaggtctt
4861 gccgcagccc gcgtagctgc aggtgtgagt ggcggtgcgt ttgcggggcc aagagcggcg
4921 gccgcgcttt ggcttggcct ccagcagctc cagcggggac gcaggcggcg tgaggagacc
4981 gcgggcggcg gggggcgcca ggcccagggc tgccgcggcg gcggccgcgt cgtcgaagag
5041 accgaaggct gggggcgcag gcggcgcgta atgcaggccg ggcccgggcg cgccgaaacc
5101 agggccaccg aaaggcggcg ggaaggaggc gcgcggaccg ggcgcgggca ggcgcgcggg
5161 gccgtcgggg ctgagcggcg gtgtgtcggg cggcggcggg gggcggcccc cgggacctcg
5221 catgcacgaa gccgccgggc ccggggcgcc ctcgcgcttg aggccgcgcg gtccacgggt
5281 cagcccgggg gcgcagccgt agccgccgcc gccgtccgct tcagggggct cggccttgac
5341 caggcggccg ggcggcgcca gcagaaagcg gccgtgcagt gcgggccccg gcggcgcatc
5401 cagctcgggt cgcagcagct cagacaccag gccacccgcg ggggcgctgt agggcggggg
5461 cgcgccgggt tcggggtaat agaacgcagg cggcgggggc ggcggcggcg gctccggggc
5521 ggcctcggcg cccaggccat ccagccccat ggacaggatg aagtccagca cgctgttgag
5581 gtcgtcgtcg gtgccgccgg actcgggttc ggcgcgcggc cagcgctcct gcaggccgcg
5641 ctcgcggcac gggctggcga aagtggagaa ggacggcagg atgggttcac tcagcgccat
5701 ggtggctagc acgcgtcagc tgactagagg atccccgggt accgagctcg aaatctccag
5761 gcgatctgac ggttcactaa acgagctctg cttatatagg cctcccaccg tacacgccta
5821 cctcgacata cgttctctat cactgatagg gagtaaactc gacatacgtt ctctatcact
5881 gatagggata aactcgacat acgttctcta tcactgatag ggagtaaact cgacatacgt
5941 tctctatcac tgatagggag taaactcgac atacgttctc tatcactgat agggagtaaa
6001 ctcgacatcg ttctctatca ctgataggga gtaaactcga catacgttct ctatcactga
6061 tagggagtaa actcgagtat gtcgaggtgg cgtgtacggt gggaggccta tataagcaga
6121 gctcgtttag tgaaccgtca gatcgcctgg agaattcgcc taggcaatta accctcacta
6181 aagggaaggc cgcctgggcc acgcgtgcca tggcgggaca cctggcttcg gatttcgcct
6241 tctcgccccc tccaggtggt ggaggtgatg ggccaggggg gccggagccg ggctgggttg
6301 atcctcggac ctggctaagc ttccaaggcc ctcctggagg gccaggaatc gggccggggg
6361 ttgggccagg ctctgaggtg tgggggattc ccccatgccc cccgccgtat gagttctgtg
6421 gggggatggc gtactgtggg ccccaggttg gagtggggct agtgccccaa ggcggcttgg
6481 agacctctca gcctgagggt gaagcaggag tcggggtgga gagcaactcc gatggggcct
6541 ccccggagcc ctgcaccgtc acccctggtg ccgtgaagct ggagaaggag aagctggagc
6601 aaaacccgga ggagtcccag gacatcaaag ctctgcagaa agaactcgag caatttgcca
6661 agctcctgaa gcagaagagg atcaccctgg gatatacaca ggccgatgtg gggctcaccc
6721 tgggggttct atttgggaag gtattcagcc aaacgaccat ctgccgcttt gaggctctgc
6781 agcttagctt caagaacatg tgtaagctgc ggcccttgct gcagaagtgg gtggaggaag
6841 ctgacaacaa tgaaaatctt caggagatat gcaaagcaga aaccctcgtg caggcccgaa
6901 agagaaagcg aaccagtatc gagaaccgag tgagaggcaa cctggagaat ttgttcctgc
6961 agtgcccgaa acccacactg cagcagatca gccacatcgc ccagcagctt gggctcgaga
7021 aggatgtggt ccgagtgtgg ttctgtaacc ggcgccagaa gggcaagcga tcaagcagcg
7081 actatgcaca acgagaggat tttgaggctg ctgggtctcc tttctcaggg ggaccagtgt
7141 cctttcctct ggccccaggg ccccattttg gtaccccagg ctatgggagc cctcacttca
7201 ctgcactgta ctcctcggtc cctttccctg agggggaagc ctttccccct gtctctgtca
7261 ccactctggg ctctcccatg cattcaaacg gggcgcgtaa aattgtcgct cctgtcaaac
7321 aaactcttaa ctttgattta ctcaaactgg ctggggatgt agaaagcaat ccaggtccag
7381 gggcgcgtgg tatggctgtc agcgacgcgc tgctcccatc tttctccacg ttcgcgtctg
7441 gcccggcggg aagggagaag acactgcgtc aagcaggtgc cccgaataac cgctggcggg
7501 aggagctctc ccacatgaag cgacttcccc cagtgcttcc cggccgcccc tatgacctgg
7561 cggcggcgac cgtggccaca gacctggaga gcggcggagc cggtgcggct tgcggcggta
7621 gcaacctggc gcccctacct cggagagaga ccgaggagtt caacgatctc ctggacctgg
7681 actttattct ctccaattcg ctgacccatc ctccggagtc agtggccgcc accgtgtcct
7741 cgtcagcgtc agcctcctct tcgtcgtcgc cgtcgagcag cggccctgcc agcgcgccct
7801 ccacctgcag cttcacctat ccgatccggg ccgggaacga cccgggcgtg gcgccgggcg
7861 gcacgggcgg aggcctcctc tatggcaggg agtccgctcc ccctccgacg gctcccttca
7921 acctggcgga catcaacgac gtgagcccct cgggcggctt cgtggccgag ctcctgcggc
7981 cagaattgga cccggtgtac attccgccgc agcagccgca gccgccaggt ggcgggctga
8041 tgggcaagtt cgtgctgaag gcgtcgctga gcgcccctgg cagcgagtac ggcagcccgt
8101 cggtcatcag cgtcagcaaa ggcagccctg acggcagcca cccggtggtg gtggcgccct
8161 acaacggcgg gccgccgcgc acgtgcccca agatcaagca ggaggcggtc tcttcgtgca
8221 cccacttggg cgctggaccc cctctcagca atggccaccg gccggctgca cacgacttcc
8281 ccctggggcg gcagctcccc agcaggacta ccccgaccct gggtcttgag gaagtgctga
8341 gcagcaggga ctgtcaccct gccctgccgc ttcctcccgg cttccatccc cacccggggc
8401 ccaattaccc atccttcctg cccgatcaga tgcagccgca agtcccgccg ctccattacc
8461 aagagctcat gccacccggt tcctgcatgc cagaggagcc caagccaaag aggggaagac
8521 gatcgtggcc ccggaaaagg accgccaccc acacttgtga ttacgcgggc tgcggcaaaa
8581 cctacacaaa gagttcccat ctcaaggcac acctgcgaac ccacacaggt gagaaacctt
8641 accactgtga ctgggacggc tgtggatgga aattcgcccg ctcagatgaa ctgaccaggc
8701 actaccgtaa acacacgggg caccgcccgt tccagtgcca aaaatgcgac cgagcatttt
8761 ccaggtcgga ccacctcgcc ttacacatga agaggcattt tggggcgcgt aaaattgtcg
8821 ctcctgtcaa acaaactctt aactttgatt tactcaaact ggctggggat gtagaaagca
8881 atccaggtcc aggggcgcgt ggtatgtaca acatgatgga gacggagctg aagccgccgg
8941 gcccgcagca aacttcgggg ggcggcggcg gcaactccac cgcggcggcg gccggcggca
9001 accagaaaaa cagcccggac cgcgtcaagc ggcccatgaa tgccttcatg gtgtggtccc
9061 gcgggcagcg gcgcaagatg gcccaggaga accccaagat gcacaactcg gagatcagca
9121 agcgcctggg cgccgagtgg aaacttttgt cggagacgga gaagcggccg ttcatcgacg
9181 aggctaagcg gctgcgagcg ctgcacatga aggagcaccc ggattataaa taccggcccc
9241 ggcggaaaac caagacgctc atgaagaagg ataagtacac gctgcccggc gggctgctgg
9301 cccccggcgg caatagcatg gcgagcgggg tcggggtggg cgccggcctg ggcgcgggcg
9361 tgaaccagcg catggacagt tacgcgcaca tgaacggctg gagcaacggc agctacagca
9421 tgatgcagga ccagctgggc tacccgcagc acccgggcct caatgcgcac ggcgcagcgc
9481 agatgcagcc catgcaccgc tacgacgtga gcgccctgca gtacaactcc atgaccagct
9541 cgcagaccta catgaacggc tcgcccacct acagcatgtc ctactcgcag cagggcaccc
9601 ctggcatggc tcttggctcc atgggttcgg tggtcaagtc cgaggccagc tccagccccc
9661 ctgtggttac ctcttcctcc cactccaggg cgccctgcca ggccggggac ctccgggaca
9721 tgatcagcat gtatctcccc ggcgccgagg tgccggaacc cgccgccccc agcagacttc
9781 acatgtccca gcactaccag agcggcccgg tgcccggcac ggccattaac ggcacactgc
9841 ccctctcaca catgtgagtt taaatctagg gcggccaatt ccgcccctct cccccccccc
9901 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
9961 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
10021 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
10081 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
10141 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
10201 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
10261 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
10321 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
10381 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
10441 gaaaaacacg atgataagct tgccacaacc cacaaggaga cgaccttcca tggcgcgtgg
10501 tatgcccctc aacgttagct tcaccaacag gaactatgac ctcgactacg actcggtgca
10561 gccgtatttc tactgcgacg aggaggagaa cttctaccag cagcagcagc agagcgagct
10621 gcagcccccg gcgcccagcg aggatatctg gaagaaattc gagctgctgc ccaccccgcc
10681 cctgtcccct agccgccgct ccgggctctg ctcgccctcc tacgttgcgg tcacaccctt
10741 ctcccttcgg ggagacaacg acggcggtgg cgggagcttc tccacggccg accagctgga
10801 gatggtgacc gagctgctgg gaggagacat ggtgaaccag agtttcatct gcgacccgga
10861 cgacgagacc ttcatcaaaa acatcatcat ccaggactgt atgtggagcg gcttctcggc
10921 cgccgccaag ctcgtctcag agaagctggc ctcctaccag gctgcgcgca aagacagcgg
10981 cagcccgaac cccgcccgcg gccacagcgt ctgctccacc tccagcttgt acctgcagga
11041 tctgagcgcc gccgcctcag agtgcatcga cccctcggtg gtcttcccct accctctcaa
11101 cgacagcagc tcgcccaagt cctgcgcctc gcaagactcc agcgccttct ctccgtcctc
11161 ggattctctg ctctcctcga cggagtcctc cccgcagggc agccccgagc ccctggtgct
11221 ccatgaggag acaccgccca ccaccagcag cgactctgag gaggaacaag aagatgagga
11281 agaaatcgat gttgtttctg tggaaaagag gcaggctcct ggcaaaaggt cagagtctgg
11341 atcaccttct gctggaggcc acagcaaacc tcctcacagc ccactggtcc tcaagaggtg
11401 ccacgtctcc acacatcagc acaactacgc agcgcctccc tccactcgga aggactatcc
11461 tgctgccaag agggtcaagt tggacagtgt cagagtcctg agacagatca gcaacaaccg
11521 aaaatgcacc agccccaggt cctcggacac cgaggagaat gtcaagaggc gaacacacaa
11581 cgtcttggag cgccagagga ggaacgagct aaaacggagc ttttttgccc tgcgtgacca
11641 gatcccggag ttggaaaaca atgaaaaggc ccccaaggta gttatcctta aaaaagccac
11701 agcatacatc ctgtccgtcc aagcagagga gcaaaagctc atttctgaag aggacttgtt
11761 gcggaaacga cgagaacagt tgaaacacaa acttgaacag ctacggaact cttgtgcgta
11821 agtttatctg cagaattccg cccccccccc ctctccctcc ccccccccta acgttactgg
11881 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt
11941 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
12001 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
12061 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
12121 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
12181 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
12241 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
12301 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa
12361 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata
12421 atatggccac aaccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
12481 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
12541 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
12601 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
12661 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
12721 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
12781 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
12841 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
12901 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
12961 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
13021 agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac
13081 cccatgcatc gatttcgaac ccggggtacc tttaagacca atgacttaca aggcagctgt
13141 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaaag
13201 aagacaagat ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg
13261 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
13321 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
13381 cttttagtca gtgtggaaaa tctctagcag aattccagca cactggcggc cgttactagt
13441 ggatcatcat aatttaaaca agcaaaacca aattaagggc cagctcattc ctcccactca
13501 tgatctatag atcctaagct tatcgatgat ctagagctcg ctgatcagcc tcgactgtgc
13561 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
13621 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
13681 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
13741 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacct
13801 gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
13861 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
13921 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
13981 tgtcgtgcca gcggatcctt tgttacttta tagaagaaat tttgagtttt tgtttttttt
14041 taataaataa ataaacataa ataaattgtt tgttgaattt attattagta tgtaagtgta
14101 aatataataa aacttaatat ctattcaaat taataaataa acctcgatat acagaccgat
14161 aaaacacatg cgtcaatttt acgcatgatt atctttaacg tacgtcacaa tatgattatc
14221 tttctagggt taaagagctc cagcttttgt tccctttagt gagggttaat ttcgagcttg
14281 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
14341 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
14401 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
14461 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
14521 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
14581 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
14641 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
14701 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
14761 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
14821 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
14881 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
14941 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
15001 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
15061 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
15121 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
15181 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
15241 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
15301 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
15361 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
15421 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
15481 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
15541 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
15601 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
15661 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
15721 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
15781 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
15841 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
15901 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
15961 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
16021 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
16081 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
16141 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
16201 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacgggaagg
16261 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
16321 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
16381 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
16441 c
//
