| 质粒名称: | CLYBL-TO-hNGN2-BSD-mApple |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 124229 |
| 存储实验室: | Michael Ward |
|
载体骨架基本信息 |
|
| 载体名称: | CLYBL targeting |
| 载体出品公司: | Clontech |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | NGN2、Neurogenin 2 |
| 物种来源: | H. sapiens (human) |
|
文献引用方法 |
|
| 材料和方法部分: | CLYBL-TO-hNGN2-BSD-mApple was a gift from Michael Ward (Addgene plasmid # 124229 ; http://n2t.net/addgene:124229 ; RRID:Addgene_124229) |
| 参考文献部分: | CRISPR Interference-Based Platform for Multimodal Genetic Screens in Human iPSC-Derived Neurons. Tian R, Gachechiladze MA, Ludwig CH, Laurie MT, Hong JY, Nathaniel D, Prabhu AV, Fernandopulle MS, Patel R, Abshari M, Ward ME, Kampmann M. Neuron. 2019 Aug 5. pii: S0896-6273(19)30640-3. doi: 10.1016/j.neuron.2019.07.014. 10.1016/j.neuron.2019.07.014 PubMed 31422865 |
买家导航
CLYBL-TO-hNGN2-BSD-mApple载体质粒基本信息
CLYBL-TO-hNGN2-BSD-mApple质粒图谱载体图谱和CLYBL-TO-hNGN2-BSD-mApple载体序列质粒序列多克隆位点信息
CLYBL-TO-hNGN2-BSD-mApple质粒载体简介
CLYBL-TO-hNGN2-BSD-mApple质粒序列载体序列
如果没有序列信息，请删除本句话LOCUS CLYBL-TO-hNGN2-BSD-mApple 12234 bp DNA circular SYN 25-AUG-2024
DEFINITION Targets the human CLYBL safe harbor locus for integration of
dox-inducible NGN2 for differentiation of iPSCs into cortical
neurons. Contains a floxed BSD-NLS-mApple selection cassette..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12234)
AUTHORS Tian R, Gachechiladze MA, Ludwig CH, Laurie MT, Hong JY, Nathaniel
D, Prabhu AV, Fernandopulle MS, Patel R, Abshari M, Ward ME,
Kampmann M
TITLE CRISPR Interference-Based Platform for Multimodal Genetic Screens in
Human iPSC-Derived Neurons.
JOURNAL Neuron. 2019 Aug 5. pii: S0896-6273(19)30640-3. doi:
10.1016/j.neuron.2019.07.014.
PUBMED 31422865
REFERENCE 2 (bases 1 to 12234)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Neuron.
2019 Aug 5. pii: S0896-6273(19)30640-3. doi:
10.1016/j.neuron.2019.07.014."
FEATURES Location/Qualifiers
source 1..12234
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
intron 660..1676
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1600..1622
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1684..1703
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 1735..2481
/codon_start=1
/product="modified rtTA protein that binds tightly to
promoters containing the tet operator in the presence of
doxycycline"
/label=Tet-On(R) 3G
/translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
DALDDFDLDMLPADALDDFDLDMLPG"
primer_bind complement(2572..2591)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 2637..2692
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(2691..2710)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind 3002..3019
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 3136..3157
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3172..3202
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3210..3226
/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 3215..3237
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3234..3250
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3234..3250
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
polyA_signal 3299..3420
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3310..3329)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 3364..3383
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
promoter complement(4288..4665)
/label=TRE3G promoter
/note="3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On(R) 3G"
primer_bind complement(4355..4379)
/label=LNCX
/note="Human CMV promoter, forward primer"
protein_bind 4424..4442
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4460..4478
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4496..4514
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4532..4550
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4568..4586
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4604..4622
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4640..4658
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
rep_origin complement(6352..6940)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6432..6451)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7111..7971)
/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 7734..7753
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
protein_bind complement(8829..8862)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
misc_feature 8863..8888
/label=SA
/note="splice acceptor site"
CDS 8912..8965
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 8960..9760
/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="GPRIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(9020..9039)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 9630..9649
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 9765..9989
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
promoter 10025..10236
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
LTR 10249..10517
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from human
T-cell leukemia virus (HTLV) type 1"
CDS 10986..11039
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 11046..11072
/codon_start=1
/product="nuclear localization signal of human c-Myc
proto-oncogene (Dang and Lee, 1988)"
/label=c-myc NLS
/translation="PAAKRVKLD"
CDS 11088..11114
/codon_start=1
/product="nuclear localization signal of human c-Myc
proto-oncogene (Dang and Lee, 1988)"
/label=c-myc NLS
/translation="PAAKRVKLD"
regulatory 11163..11172
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
protein_bind complement(11950..11983)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
primer_bind complement(11999..12016)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 12005..12229
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
481 tgggggcggg gggggggggg gggcgcgcgc cggggggggg gggggggggg gggggggggg
541 ggggggsgrg gsggrgrggk gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
961 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
1201 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
1261 ggccgcctcg ggccggggrg ggctcggggg rggggcgcgg cggcccccgg agcgccggcg
1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
1621 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
1681 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaatgt cgacatgagc
1741 cggctggaca agagcaaagt gatcaacagc gccctggaac tgctgaacgg cgtgggcatc
1801 gagggcctga ccacccggaa gctggcccag aaactgggcg tggaacagcc caccctgtac
1861 tggcacgtga agaacaagcg ggccctgctg gacgccctgc ccatcgagat gctggaccgg
1921 caccacaccc acagctgccc tctggaaggc gagagctggc aggacttcct gcggaacaac
1981 gccaagagct acagatgcgc cctgctgagc caccgggacg gcgccaaagt gcacctgggc
2041 accagaccca ccgagaagca gtacgagaca ctggaaaacc agctggcctt cctgtgccag
2101 cagggcttca gcctggaaaa cgccctgtac gccctgagcg ccgtgggcca cttcaccctg
2161 ggctgcgtgc tggaagaaca ggaacaccag gtcgccaaag aggaaagaga gacacccacc
2221 accgacagca tgccccccct gctgaagcag gccatcgagc tgttcgacag acagggcgcc
2281 gagcccgcct tcctgttcgg cctggaactg atcatctgcg gcctcgagaa gcagctgaag
2341 tgcgagagcg gcggacccac cgacgccctg gacgacttcg acctggacat gctgcccgcc
2401 gatgccctgg atgattttga tctggatatg ctccctgctg acgctctcga tgacttcgat
2461 ctcgatatgc tgccaggcta aaacgcgtga attcactcct caggtgcagg ctgcctatca
2521 gaaggtggtg actggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc
2581 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata
2641 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
2701 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc
2761 aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta
2821 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta
2881 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt
2941 acatgtgcgg ccgtcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
3001 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
3061 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
3121 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
3181 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
3241 cagctatgac catgattacg ccaagctcta gctagaggtc gacggtatac agacatgata
3301 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
3361 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
3421 ggggtgggcg atcgattcgc gagtttaaac ttacaagata caatccctgg ctatggggag
3481 gtgaggtgca tagcggtgct tgtcgggagg tgggggctgc caatagtcca tgtctgaccc
3541 ggccgggctg gcgggcgata aagtgcagct gtagggggag gtggaattgg aggacacgga
3601 ggaggacggc gcggggctgt tggtgcaact ccacgtggag gcgggcgagg ggctgtctcc
3661 gctgctgctc agggcggcgc tggcgcctcc cgggctcagc aacactgcct cggagaagag
3721 cgcccccggc aggcccccgc cgccgccccc gcagtgatcc gccaggcgca gggtctcggt
3781 gagtgcccag atgtagttgt gggcgaagcg cagggtctcg atcttggtga gcttggcgtc
3841 ctcggggaac gtggggagca cctcgcgcag cgcgtccagt gccgcgttga ggttgtgcat
3901 gcggtttcgc tcgcggttgt tggccttcag tctacgggtc ttcttgatgc gctgcaccgt
3961 ctcggccgtc ttggcgcctc gggagacggc ccgcgcccgg gaagggcgcc gtttgcaatc
4021 gtgtaccaga cccagcagcc gtgcgggccg gcagccctcc gcacccgcag ccacgccgcc
4081 ccgcgccccc tgcccggcct cagccccgcg ctgccgacgc gccccgcctg acgcgcccgg
4141 ctcctcctcc tcttcttcgt cggcgctgga tgacagcggg gtcagggccg ccaaggcggg
4201 ggaggccgat ccgagcagca ctaacacgtc ctcttcctcc ttcaactcca aggtctcgga
4261 tttgacgaac atggtggcgg cggatccttt acgagggtag gaagtggtac ggaaagttgg
4321 tataagacaa aagtgttgtg gaattgctcc aggcgatctg acggttcact aaacgagctc
4381 tgcttttata ggcgcccacc gtacacgcct aaagcttata cgttctctat cactgatagg
4441 gagtaaactg gatatacgtt ctctatcact gatagggagt aaactgtaga tacgttctct
4501 atcactgata gggagtaaac tggtcatacg ttctctatca ctgataggga gtaaactcct
4561 tatacgttct ctatcactga tagggagtaa agtctgcata cgttctctat cactgatagg
4621 gagtaaactc ttcatacgtt ctctatcact gatagggagt aaactggtac cctccgcgcc
4681 tacagctcaa gccacatccg aagggggagg gagccgggag ctgcgcgcgg ggccgccggg
4741 gggaggggtg gcaccgccca cgccgggcgg ccacgaaggg cggggcagcg ggcgcgcgcc
4801 cggcgggggg aggggccgcg cgccgcgccc gctgggaatt ggggccctag ggggagggcg
4861 gaggcgccga cgaccgcggc acttaccgtt cgcggcgtgg cgcccggtgg tccccaaggg
4921 gagggaaggg ggaggcgggg cgaggacagt gaccggagtc tcctcagcgg tggcttttct
4981 gcttggcagc ctcagcggct ggcgccaaaa ccggactccg cccacttcct cgcccctgcg
5041 gtgcgagggt gtggaatcct ccagacgctg ggggaggggg agttgggagc ttaaaaacta
5101 gtaccccttt gggaccactt tcagcagcga actctcctgt acaccagggg tcagttccac
5161 agacgcgggc caggggtggg tcattgcggc gtgaacaata atttgactag aagttgattc
5221 gggtgtttcc ggaaggggcc gagtcaatcc gccgagttgg ggcacggaaa acaaaaaggg
5281 aaggctacta agatttttct ggcgggggtt atcattggcg taactgcagg gaccacctcc
5341 cttaattaaa tctcctcatc cttccaacat aaatatattt tgggattata tcaacattca
5401 atgttactta aagtgacctt gtaaatattt tcacaactga gccatgtttg atttgtatac
5461 ttatgtttac tttactgttt ttcctgaagt taataattgc cttgaattta tttatttctt
5521 taaaaatgtt tcattactca ggactgtagt ttacattacg attctttgtg ttatacagtt
5581 gatgggtttc ttttctttct taatttcttt aaaaaataga gatggggtct tactatatta
5641 cccaggctgg tcttgaagtc ctgggctcaa gtgatcttcc tgtctcagcc taccaagtag
5701 ctgagactat aggtgcaaaa aagccactat acctggctag tttacaggtt ttaacaaatg
5761 cattatgcca cgtatccatt attacaggat cacacaagat attttcatta ccctgaaagc
5821 atccctgtgt tccaccaatt catcctgcct ccatgagccg ctggcaacca ctgatctcta
5881 tagttttgcc ttttctaaaa tgtcatataa ttggaatcat acagtctgta gcattttcag
5941 actagctttt aaaatttggc aatatgcatt taaggttcct ccttaaatgt gaagggcata
6001 gccaatgtgg cttgatagct catttctttt tattggtgaa tatttcattg tctggatgtt
6061 ccacagtttg tttatccatt cacctattca atttgctttt tttctgtgta tctatcacta
6121 attcaatact ggactctcca acagagccgt cccgcttccc taaggccctg ctctgggctt
6181 ctgggtttga gtccttggca agcccaggag aggcgctcag gcttccctgt cccccttcct
6241 cgtccaccat ctcatgcccc tggctctcct gccccttccc tacaggggtt cctggctctg
6301 ctctaagggc gaattcgcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
6361 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
6421 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
6481 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
6541 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
6601 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
6661 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
6721 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
6781 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
6841 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
6901 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
6961 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
7021 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
7081 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
7141 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
7201 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
7261 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
7321 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
7381 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
7441 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
7501 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
7561 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
7621 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
7681 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
7741 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
7801 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
7861 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
7921 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
7981 ttttcaatag caggagaatc atgccaatgg gccaatatac attctgaccc acagtttcat
8041 aataaaataa aatggttgtg gttgtaagcc actatgtttc agagtggttt gttacacagc
8101 aataaataac taatatagta ggcataccat caagtccaaa gtaggtagag aagaatgtaa
8161 atagcagagc aaaacagcat gactggtggc tgggaggctt aaaactggga caggatcaga
8221 gtcatgaaag aagtcaaaga aatggttcag aagtaaggct gagactgact tacaaaagct
8281 gaaagtccct ttaagttggt gtttggtgca ttggcagggg caggtatggt gacttaaaag
8341 agccatgctc aacaagatca agcacaacac aatcacgggt caccccagca gaccttagcg
8401 agtctagcca tttctttggt ggtggtcaca gtcatgcttc agcccagttt ccacttggac
8461 aaatggtaca tattttcaat gagatgaaaa ttaagataca atccatgtgc tcagagagtg
8521 atcacagctc tgactaaaca ctgtgcccca aagtgttgag gaattgggaa aacctagctg
8581 agttagtggt ctcttttctg ttacaataaa gctcataatg aaaattagcc ttctttgttc
8641 ttccccaagt ctttctttct agacgaaact actttcaact gttttaactt ccttactgtt
8701 aacttccata tttctagata gtatggtcag actgttattt cttgactttt aaatgtaaga
8761 tattatctac tgacttcctt ctatgtaaga tgaggattga gctctcttac ccttctccca
8821 gcggccgcat aacttcgtat agcatacatt atacgaagtt atctgacctc ttctcttcct
8881 cccacagggc ctcgagagat ctggcagcgg agagggcaga ggaagtcttc taacatgcgg
8941 tgacgtggag gagaatcccg gccctaggat tgaacaagat ggattgcacg caggttctcc
9001 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
9061 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
9121 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
9181 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
9241 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
9301 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
9361 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
9421 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
9481 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
9541 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
9601 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
9661 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
9721 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctaa aaacctgtgc cttctagttg
9781 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
9841 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
9901 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
9961 gcatgctggg gatgcggtgg gctctatgga tgcatgatct gcgatcgctc cggtgcccgt
10021 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
10081 tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
10141 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
10201 gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc
10261 tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt
10321 ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag
10381 ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg
10441 ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt
10501 ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagcgccacc atgaagacct
10561 tcaacatctc ccagcaggat ctagaattag tagaagtagc gacagagaag attacaatgc
10621 tttatgagga taataaacat catgtgggag cggcaattcg tacgaaaaca ggagaaatca
10681 tttcggcagt acatattgaa gcgtatatag gacgagtaac tgtttgtgca gaagccattg
10741 cgattggtag tgcagtttcg aatggacaaa aggattttga cacgattgta gctgttagac
10801 acccttattc tgacgaagta gatagaagta ttcgagtggt aagtccttgt ggtatgtgta
10861 gggagttgat ttcagactat gcaccagatt gttttgtgtt aatagaaatg aatggcaagt
10921 tagtcaaaac tacgattgaa gaactcattc cactcaaata tacccgaaat ggatcgggat
10981 cgggagaggg cagaggaagt ctgctaacat gcggtgacgt cgaggagaat cctggcccac
11041 cggtccctgc cgctaaacgg gtcaaactcg acggcggttc tggaggtcct gctgcaaagc
11101 gcgttaagtt ggatggaggg tcagggggag ccaccgccac cccggtcgcc acgctaccgg
11161 tcgccaccat ggtgtccaag ggcgaagaga acaacatggc catcatcaaa gaattcatgc
11221 ggttcaaggt gcacatggaa ggcagcgtga acggccacga gttcgagatt gaaggcgaag
11281 gcgagggcag accctacgag gcttttcaga ccgccaagct gaaagtgacc aaaggcggcc
11341 ctctgccttt tgcctgggac attctgagcc ctcagtttat gtacggcagc aaggtgtaca
11401 tcaagcaccc cgccgacatt cccgactact tcaagctgag cttccccgag ggcttcagat
11461 gggagagagt gatgaacttc gaggacggcg gcatcatcca cgtgaaccag gatagctctc
11521 tgcaggacgg cgtgttcatc tacaaagtga agctgcgggg caccaacttt ccctctgatg
11581 gccccgtgat gcagaaaaag accatgggct gggaagccag cgaggaaaga atgtaccctg
11641 aggatggcgc cctgaagtcc gagatcaaga agcggctgaa gctgaaggat ggcggccact
11701 atgccgccga agtgaaaacc acctacaagg ccaagaaacc cgtgcagctg cctggcgcct
11761 acatcgtgga tatcaagctg gacatcgtgt cccacaacga ggactacacc atcgtggaac
11821 agtacgagag agccgaaggc agacacagca caggcggaat ggacgagctg tacaagtaac
11881 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag
11941 taaggtacca taacttcgta tagcatacat tatacgaagt tataaacccg ctgatcagcc
12001 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
12061 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
12121 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
12181 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggg tctc
//
