| 质粒名称: | TetOn-mCherry-eGFP-RAMP4 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 109014 |
| 存储实验室: | Jacob Corn |
|
载体骨架基本信息 |
|
| 载体名称: | -- |
| 载体出品公司: | Jacob Corn |
| 空载体大小: | 7860 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，慢病毒 |
| 筛选标记: | mCherry-eGFP |
| 载体抗性: | 氨苄霉素 |
| 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: | RAMP4,SERP1 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1743 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | CMV promoter |
| 标签/融合蛋白: | mCherry eGFP（插入物上的N端） |
| 克隆方法: | Ligation Independent克隆 |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | CTCGTTTAGTGAACCGTCAGATCG |
| 3’测序引物 | CGTGAAGAATGTGCGAGACCC |
|
文献引用方法 |
|
| 材料和方法部分: | TetOn-mCherry-eGFP-RAMP4 was a gift from Jacob Corn (Addgene plasmid # 109014 ; http://n2t.net/addgene:109014 ; RRID:Addgene_109014) |
| 参考文献部分: | Atlastins remodel the endoplasmic reticulum for selective autophagy. Liang JR, Lingeman E, Ahmed S, Corn JE. J Cell Biol. 2018 Aug 24. pii: jcb.201804185. doi: 10.1083/jcb.201804185. 10.1083/jcb.201804185 PubMed 30143524 |
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 TetOn-mCherry-eGFP-RAMP4 9602 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Tet-On construct to induce expression of tandem eGFP-mCherry tagged
RAMP4 (ER marker).
ACCESSION .
VERSION .
KEYWORDS TetOn-mCherry-eGFP-RAMP4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9602)
AUTHORS Liang JR, Lingeman E, Ahmed S, Corn JE
TITLE Atlastins remodel the endoplasmic reticulum for selective autophagy.
JOURNAL J Cell Biol. 2018 Aug 24. pii: jcb.201804185. doi:
10.1083/jcb.201804185.
PUBMED 30143524
REFERENCE 2 (bases 1 to 9602)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9602
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 41..158
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 212..526
/label=tight TRE promoter
/note="Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter"
protein_bind 220..238
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 256..274
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 291..309
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 327..345
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 363..381
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 398..416
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 434..452
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 499..523
/label=LNCX
/note="Human CMV promoter, forward primer"
regulatory 586..595
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 592..1299
/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"
primer_bind complement(737..755)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1005..1024
/label=mCherry-F
/note="mCherry, forward primer"
protein_bind 1327..1351
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
regulatory 1360..1369
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1366..2082
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 1366..2082
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1411..1432)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1672..1691)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2019..2040
/label=EGFP-C
/note="EGFP, forward primer"
promoter 2341..2824
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 2770..2788
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 2860..3651
/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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2914..2933)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3524..3543
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
CDS 3661..3714
/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 3715..4461
/codon_start=1
/product="improved tetracycline-controlled transactivator"
/label=rtTA-Advanced
/note="In the Tet-On(R) system, rtTA-Advanced binds to the
Tet-responsive element and stimulates transcription only in
the presence of tetracycline or doxycycline."
/translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
DALDDFDLDMLPADALDDFDLDMLPG"
primer_bind complement(4501..4521)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4919..4930)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 5135..5368
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 5440..5561
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5477..5496)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5531..5550
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 5601..5736
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5663..5682
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 5736..5807
/gene="bla"
/label=AmpR promoter
CDS 5808..6668
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(6026..6045)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6839..7427
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7328..7347
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7581..7598
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 7715..7736
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7751..7781
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7789..7805
/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 7794..7816
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 7813..7829
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7813..7829
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 7848..7868
/label=T3
/note="T3 promoter, forward primer"
promoter 7850..7868
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 7896..8122
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 8123..8303
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 8350..8475
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 8968..9201
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 9386..9430
/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"
ORIGIN
1 aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
61 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
121 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
181 gagatccact ttggccgcga atcgatatgt cgagtttact ccctatcagt gatagagaac
241 gtatgtcgag tttactccct atcagtgata gagaacgatg tcgagtttac tccctatcag
301 tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt
361 actccctatc agtgatagag aacgtatgtc gagtttatcc ctatcagtga tagagaacgt
421 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg
481 ggaggcctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gaattggcta
541 gcgccacccc agccctcact ccttctctag gcgccggccg gatctgccac catggtgagc
601 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
661 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
721 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
781 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
841 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
901 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
961 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
1021 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
1081 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
1141 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
1201 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
1261 ggccgccact ccaccggcgg catggacgag ctgtacaagt ccggactcag atccgctggc
1321 ccaggaacaa gtttgtacaa aaaagcaggc tttccggtcg ccaccatggt gagcaagggc
1381 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
1441 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
1501 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
1561 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
1621 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
1681 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
1741 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
1801 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
1861 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
1921 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
1981 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
2041 accgccgccg ggatcactct cggcatggac gagctgtaca agtccggact cagatccggt
2101 accgaattcg gtggatctgg aggttctggt ggaatggtcg ccaagcaaag gatccgtatg
2161 gccaacgaga agcacagcaa gaacatcacc cagcgcggca acgtcgccaa gacctcgaga
2221 aatgcccccg aagagaaggc gtctgtagga ccctggttat tggctctctt catttttgtt
2281 gtctgtggtt ctgcaatttt ccagattatt caaagtatca ggatgggcat gtgaggatcc
2341 ggggttgggg ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc
2401 tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc
2461 cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc
2521 tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac
2581 ggaagccgca cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc
2641 gcgccgaccg cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag
2701 cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct
2761 gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct
2821 cgttgaccga atcaccgacc tcttccccag caattcacca tgattgaaca agatggattg
2881 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
2941 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
3001 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
3061 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
3121 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
3181 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
3241 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
3301 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
3361 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
3421 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
3481 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
3541 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
3601 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ccccggtgcc
3661 gaaggtagag gttctctcct cacttgtggt gatgttgaag aaaaccctgg tccaatgtct
3721 agactggaca agagcaaagt cataaacgga gctctggaat tactcaatgg tgtcggtatc
3781 gaaggcctga cgacaaggaa actcgctcaa aagctgggag ttgagcagcc taccctgtac
3841 tggcacgtga agaacaagcg ggccctgctc gatgccctgc caatcgagat gctggacagg
3901 catcataccc acttctgccc cctggaaggc gagtcatggc aagactttct gcggaacaac
3961 gccaagtcat accgctgtgc tctcctctca catcgcgacg gggctaaagt gcatctcggc
4021 acccgcccaa cagagaaaca gtacgaaacc ctggaaaatc agctcgcgtt cctgtgtcag
4081 caaggcttct ccctggagaa cgcactgtac gctctgtccg ccgtgggcca ctttacactg
4141 ggctgcgtat tggaggaaca ggagcatcaa gtagcaaaag aggaaagaga gacacctacc
4201 accgattcta tgcccccact tctgagacaa gcaattgagc tgttcgaccg gcagggagcc
4261 gaacctgcct tccttttcgg cctggaacta atcatatgtg gcctggagaa acagctaaag
4321 tgcgaaagcg gcgggccgac cgacgccctt gacgattttg acttagacat gctcccagcc
4381 gatgcccttg acgactttga ccttgatatg ctgcctgctg acgctcttga cgattttgac
4441 cttgacatgc tccccgggta actaagtaag gatcgatcca agatatcgta ttcttaacta
4501 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
4561 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
4621 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
4681 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
4741 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
4801 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
4861 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
4921 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
4981 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgt
5041 ttcgcctcgg cgtccggact agaggtacct ttaagaccaa tgacttacaa ggcagctgta
5101 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
5161 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
5221 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
5281 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
5341 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
5401 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat
5461 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
5521 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta
5581 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
5641 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
5701 tattccagaa gtagtgagga ggcttttttg gaggcctttc aaatatgtat ccgctcatga
5761 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
5821 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
5881 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
5941 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
6001 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
6061 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
6121 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
6181 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
6241 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
6301 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
6361 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
6421 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
6481 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
6541 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
6601 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
6661 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
6721 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
6781 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
6841 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
6901 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
6961 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
7021 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
7081 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
7141 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
7201 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
7261 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
7321 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
7381 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
7441 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
7501 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
7561 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
7621 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
7681 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
7741 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
7801 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
7861 actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta
7921 gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg
7981 tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac
8041 gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa
8101 gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct
8161 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
8221 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
8281 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
8341 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
8401 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
8461 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
8521 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
8581 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
8641 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
8701 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
8761 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct
8821 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata
8881 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt
8941 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc
9001 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc
9061 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt
9121 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata
9181 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac
9241 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac
9301 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat
9361 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg
9421 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat
9481 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa
9541 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg
9601 ac
//
