| 质粒名称: | pHAGE-TO-nmdCas9-3XmCherry |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 64110 |
| 存储实验室: | Thoru Pederson |
|
载体骨架基本信息 |
|
| 载体名称: | pHAGE-DEST |
| 载体出品公司: | Clontech |
| 空载体大小: | 6000bp |
| 完整质粒大小: | 12104bp |
| 载体类型: | 哺乳细胞表达, Lentiviral, CRISPR |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | Stbl3 |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Nm dCas9 |
| 物种来源: | Synthetic |
| 插入大小: | 3243bp |
| 启动子: | CMV-TO |
| 标签/融合蛋白: | 3XmCherry (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI (not destroyed) |
| 3’克隆位点: | XbaI (not destroyed) |
| 5’测序引物 | cgaaggaatagaagaagaaggtgga |
| 3’测序引物 |
CCAAAGGGAGATCCGACTCG
(Common Sequencing Primers)
|
|
文献引用方法 |
|
| 材料和方法部分: | pHAGE-TO-nmdCas9-3XmCherry was a gift from Thoru Pederson (Addgene plasmid # 64110 ; http://n2t.net/addgene:64110 ; RRID:Addgene_64110) |
| 参考文献部分: | Multicolor CRISPR labeling of chromosomal loci in human cells. Ma H, Naseri A, Reyes-Gutierrez P, Wolfe SA, Zhang S, Pederson T. Proc Natl Acad Sci U S A. 2015 Mar 10;112(10):3002-7. doi: 10.1073/pnas.1420024112. Epub 2015 Feb 23. 10.1073/pnas.1420024112 PubMed 25713381 |
买家导航
pHAGE-TO-nmdCas9-3XmCherry载体质粒基本信息
pHAGE-TO-nmdCas9-3XmCherry质粒图谱载体图谱和pHAGE-TO-nmdCas9-3XmCherry载体序列质粒序列多克隆位点信息
pHAGE-TO-nmdCas9-3XmCherry质粒载体简介
pHAGE-TO-nmdCas9-3XmCherry质粒序列载体序列
LOCUS pHAGE-TO-nmdCas9-3XmCherry 12104 bp DNA circular SYN 14-APR-2025
DEFINITION Expression of Nm dCas9-3XmCherry in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12104)
AUTHORS Ma H, Naseri A, Reyes-Gutierrez P, Wolfe SA, Zhang S, Pederson T
TITLE Multicolor CRISPR labeling of chromosomal loci in human cells.
JOURNAL Proc Natl Acad Sci U S A. 2015 Mar 10;112(10):3002-7. doi:
10.1073/pnas.1420024112. Epub 2015 Feb 23.
PUBMED 25713381
REFERENCE 2 (bases 1 to 12104)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1073/pnas.1420024112"; journalName: "Proc Natl Acad Sci U S A";
date: "2015-03-10- 10"; volume: "112"; issue: "10"; pages: "3002-7"
COMMENT Sequence Label: pHAGE-TO-nmdCas9-3XmCherry
FEATURES Location/Qualifiers
source 1..12104
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..204
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 155..175
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
protein_bind 206..224
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 227..245
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 255..279
/label=LNCX
/note="Human CMV promoter, forward primer"
regulatory 372..381
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 384..404
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS join(417..458,459..461,462..2171,2172..2177,2178..2243,
2244..2246,2247..3659)
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Neisseria meningitidis Type II
CRISPR/Cas system"
/label=NmdCas9
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the mutations D16A, D587A,
H588A, and N611A"
/translation="AAFKPNPINYILGLAIGIASVGWAMVEIDEDENPICLIDLGVRVF
ERAEVPKTGDSLAMARRLARSVRRLTRRRAHRLLRARRLLKREGVLQAADFDENGLIKS
LPNTPWQLRAAALDRKLTPLEWSAVLLHLIKHRGYLSQRKNEGETADKELGALLKGVAD
NAHALQTGDFRTPAELALNKFEKESGHIRNQRGDYSHTFSRKDLQAELILLFEKQKEFG
NPHVSGGLKEGIETLLMTQRPALSGDAVQKMLGHCTFEPAEPKAAKNTYTAERFIWLTK
LNNLRILEQGSERPLTDTERATLMDEPYRKSKLTYAQARKLLGLEDTAFFKGLRYGKDN
AEASTLMEMKAYHAISRALEKEGLKDKKSPLNLSPELQDEIGTAFSLFKTDEDITGRLK
DRIQPEILEALLKHISFDKFVQISLKALRRIVPLMEQGKRYDEACAEIYGDHYGKKNTE
EKIYLPPIPADEIRNPVVLRALSQARKVINGVVRRYGSPARIHIETAREVGKSFKDRKE
IEKRQEENRKDREKAAAKFREYFPNFVGEPKSKDILKLRLYEQQHGKCLYSGKEINLGR
LNEKGYVEIAAALPFSRTWDDSFNNKVLVLGSEAQNKGNQTPYEYFNGKDNSREWQEFK
ARVETSRFPRSKKQRILLQKFDEDGFKERNLNDTRYVNRFLCQFVADRMRLTGKGKKRV
FASNGQITNLLRGFWGLRKVRAENDRHHALDAVVVACSTVAMQQKITRFVRYKEMNAFD
GKTIDKETGEVLHQKTHFPQPWEFFAQEVMIRVFGKPDGKPEFEEADTPEKLRTLLAEK
LSSRPEAVHEYVTPLFVSRAPNRKMSGQGHMETVKSAKRLDEGVSVLRVPLTQLKLKDL
EKMVNREREPKLYEALKARLEAHKDDPAKAFAEPFYKYDKAGNRTQQVKAVRVEQVQKT
GVWVRNHNGIADNATMVRVDVFEKGDKYYLVPIYSWQVAKGILPDRAVVQGKDEEDWQL
IDDSFNFKFSLHPNDLVEVITKKARMFGYFASCHRGTGNINIRIHDLDHKIGKNGILEG
IGVKTALSFQKYQIDELGKEIRPCRLKKRPPVR"
regulatory 3246..3255
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 3678..3698
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 3711..4418
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
CDS 4437..5144
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
CDS 5163..5870
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
misc_feature 5892..6480
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5945..5965)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6363..6374)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 6555..6788
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(6907..6925)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6993..7097
/gene="bla"
/label=AmpR promoter
CDS join(7098..7166,7167..7958)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(7316..7335)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8129..8717
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8618..8637
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8871..8888
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 9128..9149
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter join(9164..9169,9170..9187,9188..9194)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9202..9218
/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 9207..9229
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 9226..9242
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 9226..9242
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 9292..9311
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
LTR 9503..10136
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 10183..10308
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 10805..11038
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 11223..11267
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 11560..11677
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 11801..12104
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
181 tgggaggtct atataagcag agctctccct atcagtgata gagatctccc tatcagtgat
241 agagatcgtc gacgagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc
301 tgttttgacc tccatagaag acaccgggac cgatccagcc taccggttaa ggtgaacgcg
361 tgcaggctgg cgccaccatg gcccctaaaa agaaacgcaa ggtcgaattc ggaagcgccg
421 ccttcaagcc caaccccatc aactacatcc tgggcctggc catcggcatc gccagcgtgg
481 gctgggccat ggtggagatc gacgaggacg agaaccccat ctgcctgatc gacctgggtg
541 tgcgcgtgtt cgagcgcgct gaggtgccca agactggtga cagtctggct atggctcgcc
601 ggcttgctcg ctctgttcgg cgccttactc gccggcgcgc tcaccgcctt ctgcgcgctc
661 gccgcctgct gaagcgcgag ggtgtgctgc aggctgccga cttcgacgag aacggcctga
721 tcaagagcct gcccaacact ccttggcagc tgcgcgctgc cgctctggac cgcaagctga
781 ctcctctgga gtggagcgcc gtgctgctgc acctgatcaa gcaccgcggc tacctgagcc
841 agcgcaagaa cgagggcgag accgccgaca aggagctggg tgctctgctg aagggcgtgg
901 ccgacaacgc ccacgccctg cagactggtg acttccgcac tcctgctgag ctggccctga
961 acaagttcga gaaggagagc ggccacatcc gcaaccagcg cggcgactac agccacacct
1021 tcagccgcaa ggacctgcag gccgagctga tcctgctgtt cgagaagcag aaggagttcg
1081 gcaaccccca cgtgagcggc ggcctgaagg agggcatcga gaccctgctg atgacccagc
1141 gccccgccct gagcggcgac gccgtgcaga agatgctggg ccactgcacc ttcgagccag
1201 ccgagcccaa ggccgccaag aacacctaca ccgccgagcg cttcatctgg ctgaccaagc
1261 tgaacaacct gcgcatcctg gagcagggca gcgagcgccc cctgaccgac accgagcgcg
1321 ccaccctgat ggacgagccc taccgcaaga gcaagctgac ctacgcccag gcccgcaagc
1381 tgctgggtct ggaggacacc gccttcttca agggcctgcg ctacggcaag gacaacgccg
1441 aggccagcac cctgatggag atgaaggcct accacgccat cagccgcgcc ctggagaagg
1501 agggcctgaa ggacaagaag agtcctctga acctgagccc cgagctgcag gacgagatcg
1561 gcaccgcctt cagcctgttc aagaccgacg aggacatcac cggccgcctg aaggaccgca
1621 tccagcccga gatcctggag gccctgctga agcacatcag cttcgacaag ttcgtgcaga
1681 tcagcctgaa ggccctgcgc cgcatcgtgc ccctgatgga gcagggcaag cgctacgacg
1741 aggcctgcgc cgagatctac ggcgaccact acggcaagaa gaacaccgag gagaagatct
1801 acctgcctcc tatccccgcc gacgagatcc gcaaccccgt ggtgctgcgc gccctgagcc
1861 aggcccgcaa ggtgatcaac ggcgtggtgc gccgctacgg cagccccgcc cgcatccaca
1921 tcgagaccgc ccgcgaggtg ggcaagagct tcaaggaccg caaggagatc gagaagcgcc
1981 aggaggagaa ccgcaaggac cgcgagaagg ccgccgccaa gttccgcgag tacttcccca
2041 acttcgtggg cgagcccaag agcaaggaca tcctgaagct gcgcctgtac gagcagcagc
2101 acggcaagtg cctgtacagc ggcaaggaga tcaacctggg ccgcctgaac gagaagggct
2161 acgtggagat cgccgctgcc ctgcccttca gccgcacctg ggacgacagc ttcaacaaca
2221 aggtgctggt gctgggcagc gaggctcaga acaagggcaa ccagaccccc tacgagtact
2281 tcaacggcaa ggacaacagc cgcgagtggc aggagttcaa ggcccgcgtg gagaccagcc
2341 gcttcccccg cagcaagaag cagcgcatcc tgctgcagaa gttcgacgag gacggcttca
2401 aggagcgcaa cctgaacgac acccgctacg tgaaccgctt cctgtgccag ttcgtggccg
2461 accgcatgcg cctgaccggc aagggcaaga agcgcgtgtt cgccagcaac ggccagatca
2521 ccaacctgct gcgcggcttc tggggcctgc gcaaggtgcg cgccgagaac gaccgccacc
2581 acgccctgga cgccgtggtg gtggcctgca gcaccgtggc catgcagcag aagatcaccc
2641 gcttcgtgcg ctacaaggag atgaacgcct tcgacggtaa aaccatcgac aaggagaccg
2701 gcgaggtgct gcaccagaag acccacttcc cccagccctg ggagttcttc gcccaggagg
2761 tgatgatccg cgtgttcggc aagcccgacg gcaagcccga gttcgaggag gccgacaccc
2821 ccgagaagct gcgcaccctg ctggccgaga agctgagcag ccgccctgag gccgtgcacg
2881 agtacgtgac tcctctgttc gtgagccgcg cccccaaccg caagatgagc ggtcagggtc
2941 acatggagac cgtgaagagc gccaagcgcc tggacgaggg cgtgagcgtg ctgcgcgtgc
3001 ccctgaccca gctgaagctg aaggacctgg agaagatggt gaaccgcgag cgcgagccca
3061 agctgtacga ggccctgaag gcccgcctgg aggcccacaa ggacgacccc gccaaggcct
3121 tcgccgagcc cttctacaag tacgacaagg ccggcaaccg cacccagcag gtgaaggccg
3181 tgcgcgtgga gcaggtgcag aagaccggcg tgtgggtgcg caaccacaac ggcatcgccg
3241 acaacgccac catggtgcgc gtggacgtgt tcgagaaggg cgacaagtac tacctggtgc
3301 ccatctacag ctggcaggtg gccaagggca tcctgcccga ccgcgccgtg gtgcagggca
3361 aggacgagga ggactggcag ctgatcgacg acagcttcaa cttcaagttc agcctgcacc
3421 ccaacgacct ggtggaggtg atcaccaaga aggcccgcat gttcggctac ttcgccagct
3481 gccaccgcgg caccggcaac atcaacatcc gcatccacga cctggaccac aagatcggca
3541 agaacggcat cctggagggc atcggcgtga agaccgccct gagcttccag aagtaccaga
3601 tcgacgagct gggcaaggag atccgcccct gccgcctgaa gaagcgccct cctgtgcgcg
3661 gctctactag tggctctccc aagaagaaga ggaaggtagg atccggaagt atggtgagca
3721 agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg
3781 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
3841 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg
3901 acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca
3961 tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
4021 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc actgcaggac ggcgagttca
4081 tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga
4141 agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg
4201 gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga
4261 ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt
4321 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgctgagg
4381 gccgccactc caccggcggc atggacgagc tgtacaaggg tagcatcgat ggtagcatgg
4441 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
4501 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
4561 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
4621 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
4681 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
4741 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctcactg caggacggcg
4801 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
4861 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
4921 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
4981 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
5041 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
5101 ctgagggccg ccactccacc ggcggcatgg acgagctgta caagtctggc ggccgctctg
5161 gcatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
5221 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
5281 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
5341 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
5401 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
5461 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tcactgcagg
5521 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
5581 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
5641 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
5701 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
5761 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
5821 aacgcgctga gggccgccac tccaccggcg gcatggacga gctgtacaag ctcgagtagg
5881 gtaatctaga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
5941 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
6001 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
6061 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
6121 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
6181 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
6241 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
6301 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
6361 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
6421 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
6481 ctgagatcct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
6541 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc
6601 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
6661 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
6721 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
6781 ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
6841 aatatcagag agtgagaggc ccgggttaat taaggaaagg gctagatcat tcttgaagac
6901 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
6961 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
7021 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
7081 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
7141 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
7201 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
7261 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
7321 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
7381 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
7441 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
7501 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
7561 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
7621 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
7681 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
7741 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
7801 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
7861 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
7921 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
7981 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
8041 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
8101 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
8161 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
8221 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
8281 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
8341 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
8401 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
8461 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
8521 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
8581 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
8641 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
8701 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
8761 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
8821 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
8881 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
8941 ttcattaatg cagcaagctc atggctgact aatttttttt atttatgcag aggccgaggc
9001 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
9061 ttgcaaaaag ctccccgtgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
9121 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
9181 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
9241 acatgattac gaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
9301 ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa
9361 atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt
9421 tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa
9481 tatgtactac aaacttagta gttggaaggg ctaattcact cccaaagaag acaagatatc
9541 cttgatctgt ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca
9601 gggccagggg tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag
9661 ccagataagg tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc
9721 ctgcatggga tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta
9781 gcatttcatc acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga
9841 gcttgctaca agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact
9901 ggggagtggc gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc
9961 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
10021 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
10081 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
10141 cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact
10201 cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa
10261 attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg
10321 gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat
10381 aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc
10441 ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag
10501 acaggatcag aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat
10561 caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac
10621 aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat cttcagacct ggaggaggag
10681 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
10741 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
10801 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
10861 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
10921 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
10981 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
11041 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
11101 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
11161 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
11221 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
11281 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
11341 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
11401 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
11461 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac
11521 ggtatcgccg aattgacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
11581 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
11641 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
11701 agatccagtt tggactagcg atgctagctg atgcgggccc tagttattaa tagtaatcaa
11761 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
11821 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
11881 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
11941 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
12001 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
12061 ctacttggca gtacatctac gtattagtca tcgctattac catg
//
