| 质粒名称: | pLenti-EML4-ALK variant 3a |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 183829 |
| 存储实验室: | Aaron Hata |
|
载体骨架基本信息 |
|
| 载体名称: | pLenti |
| 空载体大小: | 7877 bp |
| 完整质粒大小: | 10235 bp |
| 载体类型: | 哺乳细胞表达、Lentiviral |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称1: | ALK Receptor Tyrosine Kinase / ALK |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1691 bp |
| GenBank编号: | NM_004304 |
| 启动子: | CMV |
| 标签/融合蛋白: | The fusion protein of EML4 exon 1-6a and ALK exon 20-29 |
| 克隆方法: | TOPO Cloning |
| 5’测序引物 | T7 |
| 插入基因名称2: | EMAP Like 4 / EML4 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 667bp |
| GenBank编号: | NM_019063.5 |
| 启动子: | CMV |
| 标签/融合蛋白: | The fusion protein of EML4 exon 1-6a and ALK exon 20-29 |
| 克隆方法: | TOPO Cloning |
| 5’测序引物 | T7 |
|
文献引用方法 |
|
| 材料和方法部分: | pLenti-EML4-ALK variant 3a was a gift from Aaron Hata (Addgene plasmid # 183829 ; http://n2t.net/addgene:183829 ; RRID:Addgene_183829) |
| 参考文献部分: | Sequential ALK Inhibitors Can Select for Lorlatinib-Resistant Compound ALK Mutations in ALK-Positive Lung Cancer. Yoda S, Lin JJ, Lawrence MS, Burke BJ, Friboulet L, Langenbucher A, Dardaei L, Prutisto-Chang K, Dagogo-Jack I, Timofeevski S, Hubbeling H, Gainor JF, Ferris LA, Riley AK, Kattermann KE, Timonina D, Heist RS, Iafrate AJ, Benes CH, Lennerz JK, Mino-Kenudson M, Engelman JA, Johnson TW, Hata AN, Shaw AT. Cancer Discov. 2018 Jun;8(6):714-729. doi: 10.1158/2159-8290.CD-17-1256. Epub 2018 Apr 12. 10.1158/2159-8290.CD-17-1256 PubMed 29650534 |
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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 pLenti-EML4-ALK variant 3a 10235 bp ds-DNA circular SYN 28-APR-2022
DEFINITION Express EML4-ALK fusion variant 3a (E6a;A20) in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pLenti-EML4-ALK variant 3a
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10235)
AUTHORS Yoda S, Lin JJ, Lawrence MS, Burke BJ, Friboulet L, Langenbucher A,
Dardaei L, Prutisto-Chang K, Dagogo-Jack I, Timofeevski S, Hubbeling
H, Gainor JF, Ferris LA, Riley AK, Kattermann KE, Timonina D, Heist
RS, Iafrate AJ, Benes CH, Lennerz JK, Mino-Kenudson M, Engelman JA,
Johnson TW, Hata AN, Shaw AT
TITLE Sequential ALK Inhibitors Can Select for Lorlatinib-Resistant
Compound ALK Mutations in ALK-Positive Lung Cancer.
JOURNAL Cancer Discov. 2018 Jun;8(6):714-729. doi:
10.1158/2159-8290.CD-17-1256. Epub 2018 Apr 12.
PUBMED 29650534
REFERENCE 2 (bases 1 to 10235)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 28, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..10235
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 83..103
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 129..153
/label=LNCX
/note="Human CMV promoter, forward primer"
primer_bind 228..247
/label=T7
/note="T7 promoter, forward primer"
promoter 228..246
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 276..300
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(2695..2719)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 2751..2774
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 2790..2816
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
misc_feature 2839..3391
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(2983..3000)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 3210..3229
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 3403..4002
/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(3403..3422)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3899..3919
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 4019..4607
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4072..4092)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4490..4501)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 4682..4915
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(5034..5052)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5120..5224
/gene="bla"
/label=AmpR promoter
CDS 5225..6085
/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(5443..5462)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6256..6844
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6745..6764
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6998..7015
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 7107..7126
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
protein_bind 7255..7276
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7291..7321
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7329..7345
/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 7334..7356
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 7353..7369
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7353..7369
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 7419..7438
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
LTR 7630..8263
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 8310..8435
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 8932..9165
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 9350..9394
/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 9691..9808
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 9860..10163
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 10164..132
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
ORIGIN
1 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
61 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
121 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
181 gacctccata gaagacaccg gcggccgcag atctctcgag gttgatctaa tacgactcac
241 tatagggaga cccaagctgg ctagttaagc tatcaacaag tttgtacaaa aaagcaggct
301 ccgcggccgc ccccttcacc atggacggtt tcgccggcag tctcgatgat agtatttctg
361 ctgcaagtac ttctgatgtt caagatcgcc tgtcagctct tgagtcacga gttcagcaac
421 aagaagatga aatcactgtg ctaaaggcgg ctttggctga tgttttgagg cgtcttgcaa
481 tctctgaaga tcatgtggcc tcagtgaaaa aatcagtctc aagtaaaggc caaccaagcc
541 ctcgagcagt tattcccatg tcctgtataa ccaatggaag tggtgcaaac agaaaaccaa
601 gtcataccag tgctgtctca attgcaggaa aagaaactct ttcatctgct gctaaaagtg
661 gtacagaaaa aaagaaagaa aaaccacaag gacagagaga aaaaaaagag gaatctcatt
721 ctaatgatca aagtccacaa attcgagcat caccttctcc ccagccctct tcacaacctc
781 tccaaataca cagacaaact ccagaaagca agaatgctac tcccaccaaa agcataaaac
841 gaccatcacc agctgaaaag tcacataatt cttgggaaaa ttcagatgat agccgtaata
901 aattgtcgaa aataccttca acacccaaat taataccaaa agttaccaaa actgcagaca
961 agcataaaga tgtcatcatc aaccaagtgt accgccggaa gcaccaggag ctgcaagcca
1021 tgcagatgga gctgcagagc cctgagtaca agctgagcaa gctccgcacc tcgaccatca
1081 tgaccgacta caaccccaac tactgctttg ctggcaagac ctcctccatc agtgacctga
1141 aggaggtgcc gcggaaaaac atcaccctca ttcggggtct gggccatggc gcctttgggg
1201 aggtgtatga aggccaggtg tccggaatgc ccaacgaccc aagccccctg caagtggctg
1261 tgaagacgct gcctgaagtg tgctctgaac aggacgaact ggatttcctc atggaagccc
1321 tgatcatcag caaattcaac caccagaaca ttgttcgctg cattggggtg agcctgcaat
1381 ccctgccccg gttcatcctg ctggagctca tggcgggggg agacctcaag tccttcctcc
1441 gagagacccg ccctcgcccg agccagccct cctccctggc catgctggac cttctgcacg
1501 tggctcggga cattgcctgt ggctgtcagt atttggagga aaaccacttc atccaccgag
1561 acattgctgc cagaaactgc ctcttgacct gtccaggccc tggaagagtg gccaagattg
1621 gagacttcgg gatggcccga gacatctaca gggcgagcta ctatagaaag ggaggctgtg
1681 ccatgctgcc agttaagtgg atgcccccag aggccttcat ggaaggaata ttcacttcta
1741 aaacagacac atggtccttt ggagtgctgc tatgggaaat cttttctctt ggatatatgc
1801 cataccccag caaaagcaac caggaagttc tggagtttgt caccagtgga ggccggatgg
1861 acccacccaa gaactgccct gggcctgtat accggataat gactcagtgc tggcaacatc
1921 agcctgaaga caggcccaac tttgccatca ttttggagag gattgaatac tgcacccagg
1981 acccggatgt aatcaacacc gctttgccga tagaatatgg tccacttgtg gaagaggaag
2041 agaaagtgcc tgtgaggccc aaggaccctg agggggttcc tcctctcctg gtctctcaac
2101 aggcaaaacg ggaggaggag cgcagcccag ctgccccacc acctctgcct accacctcct
2161 ctggcaaggc tgcaaagaaa cccacagctg cagaggtctc tgttcgagtc cctagagggc
2221 cggccgtgga agggggacac gtgaatatgg cattctctca gtccaaccct ccttcggagt
2281 tgcacaaggt ccacggatcc agaaacaagc ccaccagctt gtggaaccca acgtacggct
2341 cctggtttac agagaaaccc accaaaaaga ataatcctat agcaaagaag gagccacacg
2401 acaggggtaa cctggggctg gagggaagct gtactgtccc acctaacgtt gcaactggga
2461 gacttccggg ggcctcactg ctcctagagc cctcttcgct gactgccaat atgaaggagg
2521 tacctctgtt caggctacgt cacttccctt gtgggaatgt caattacggc taccagcaac
2581 agggcttgcc cttagaagcc gctactgccc ctggagctgg tcattacgag gataccattc
2641 tgaaaagcaa gaatagcatg aaccagcctg ggccctgaaa gggtgggcgc gccgacccag
2701 ctttcttgta caaagtggtt tagctagagg gcccgcggtt cgaactcgag gactacaagg
2761 acgacgatga caagtcggcc gctggaggat acccctacga cgtgcccgac tacgcctaga
2821 attccgcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
2881 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
2941 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
3001 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
3061 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
3121 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
3181 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
3241 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
3301 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
3361 tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatgaccga gtacaagccc
3421 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
3481 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
3541 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
3601 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
3661 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
3721 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
3781 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
3841 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
3901 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
3961 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gatgcatcga tggatcctaa
4021 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
4081 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
4141 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
4201 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
4261 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
4321 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
4381 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
4441 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
4501 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
4561 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg agatccttta
4621 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
4681 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
4741 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
4801 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
4861 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
4921 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
4981 gagaggcccg ggttaattaa ggaaagggct agatcattct tgaagacgaa agggcctcgt
5041 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
5101 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
5161 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
5221 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
5281 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
5341 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
5401 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
5461 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
5521 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
5581 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
5641 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
5701 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
5761 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
5821 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
5881 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
5941 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
6001 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
6061 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
6121 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
6181 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
6241 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
6301 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
6361 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
6421 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
6481 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
6541 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
6601 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
6661 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
6721 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
6781 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
6841 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
6901 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
6961 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
7021 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
7081 caagctcatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
7141 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
7201 cccgtggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
7261 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
7321 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa
7381 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
7441 tgtatcttat catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact
7501 tcccacccca taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg
7561 tgattcctct gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa
7621 cttagtagtt ggaagggcta attcactccc aaagaagaca agatatcctt gatctgtgga
7681 tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca
7741 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag
7801 aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg
7861 atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg
7921 tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg
7981 gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag
8041 ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac
8101 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
8161 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
8221 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
8281 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
8341 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
8401 gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat
8461 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata
8521 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat
8581 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag
8641 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga
8701 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca
8761 ccgcacagca agcggccggc cgctgatctt cagacctgga ggaggagata tgagggacaa
8821 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
8881 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
8941 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
9001 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
9061 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
9121 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
9181 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
9241 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
9301 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
9361 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
9421 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
9481 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
9541 gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg
9601 agagagagac agagacagat ccattcgatt agtgaacgga tctcgacggt atcgccgaat
9661 taattcacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta
9721 cagtgcaggg aaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
9781 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt
9841 ttggactagt ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
9901 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
9961 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
10021 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
10081 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
10141 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
10201 agcggtttga ctcacgggga tttccaagtc tccac
//
