| 质粒名称: | pLentipuro3/TO/V5-GW/EGFP-Firefly Luciferase |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 119816 |
| 存储实验室: | Ethan Abel |
|
载体骨架基本信息 |
|
| 载体名称: | pLentipuro3/TO/V5-DEST |
| 载体出品公司: | Clontech |
| 空载体大小: | 8116bp |
| 完整质粒大小: | 10489 bp |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 载体类型: | 哺乳细胞表达\Lentiviral, Luciferase |
| 筛选标记: | Puromycin |
| 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1: | Luciferase / Firefly luciferase |
| 物种来源: | Photinus pyralis |
| 插入大小: | 2373 bp |
| 基因突变: | Stop codon removed in the frame of the V5 frame vector |
| 启动子: | CMV/TO |
| 标签/融合蛋白: |
EGFP (N terminal on insert)
V5 (C terminal on insert)
|
| 克隆方法： | Gateway Cloning |
| 5’测序引物： | CMV-F |
| 3’测序引物： | Gateway Cloning |
| 插入基因名称2: | Puromycin |
| 物种来源: | Streptomyces alboniger |
| 插入大小: | 600bp |
| 启动子: | Human Phosphoglycerate kinase |
| 克隆方法： | 未知 |
| 5’测序引物： | 未知 |
| 3’测序引物： | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | pLentipuro3/TO/V5-GW/EGFP-Firefly Luciferase was a gift from Ethan Abel (Addgene plasmid # 119816 ; http://n2t.net/addgene:119816 ; RRID:Addgene_119816) |
| 参考文献部分: | A surgical orthotopic approach for studying the invasive progression of human bladder cancer. Lorenzatti Hiles G, Cates AL, El-Sawy L, Day KC, Broses LJ, Han AL, Briggs HL, Emamdjomeh A, Chou A, Abel EV, Liebert M, Palmbos PL, Udager AM, Keller ET, Day ML. Nat Protoc. 2019 Mar;14(3):738-755. doi: 10.1038/s41596-018-0112-8. 10.1038/s41596-018-0112-8 PubMed 30683938 |
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 pLentipuro3/TO/V5-GW/EGFP-Firefly Luciferase 10490 bp ds-DNA circular SYN 23-SEP-2022
DEFINITION 3rd generation lentiviral plasmid for expression of EGFP Firefly
Luciferase fusion protein in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pLentipuro3/TO/V5-GW/EGFP-Firefly Luciferase
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10490)
AUTHORS Lorenzatti Hiles G, Cates AL, El-Sawy L, Day KC, Broses LJ, Han AL,
Briggs HL, Emamdjomeh A, Chou A, Abel EV, Liebert M, Palmbos PL,
Udager AM, Keller ET, Day ML
TITLE A surgical orthotopic approach for studying the invasive progression
of human bladder cancer.
JOURNAL Nat Protoc. 2019 Mar;14(3):738-755. doi: 10.1038/s41596-018-0112-8.
PUBMED 30683938
REFERENCE 2 (bases 1 to 10490)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 23, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..10490
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(35..52)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(206..794)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(286..305)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(965..1825)
/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 1588..1607
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1826..1930)
/gene="bla"
/label=AmpR promoter
rep_origin complement(1956..2411)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2093..2114)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2305..2324
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2538..2560
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2552..2569
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2553..2569
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2579..2598
/label=T7
/note="T7 promoter, forward primer"
promoter 2579..2597
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin 2618..2753
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(2672..2691)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 2793..2914
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2804..2823)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2858..2877
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
LTR 2986..3219
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
misc_feature 3299..3887
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS 3405..3416
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind 3814..3834
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3955..4554)
/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(4038..4058)
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
primer_bind 4535..4554
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
promoter complement(4576..5086)
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind complement(4639..4657)
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS complement(5146..5187)
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
protein_bind 5240..5264
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS complement(5281..6930)
/codon_start=1
/gene="luc+"
/product="firefly luciferase"
/label=luciferase
/note="enhanced luc+ version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
primer_bind 6846..6864
/label=LucNrev
/note="Firefly luciferase, reverse primer"
CDS complement(6937..7653)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS complement(6937..7653)
/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(6979..7000)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 7328..7347
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 7587..7608
/label=EGFP-N
/note="EGFP, reverse primer"
protein_bind complement(7674..7698)
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind 7739..7757
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 7760..7778
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
promoter complement(7780..7983)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(7809..7829)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
misc_feature 8309..8426
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(8693..8737)
/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 8922..9155
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 9648..9773
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
LTR 9820..10000
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(10001..10227)
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
primer_bind complement(10255..10275)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(10255..10273)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(10294..10310)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(10294..10310)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(10307..10329)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 10318..10334
/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(10342..10372)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 10387..10408
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
ORIGIN
1 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
61 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
1021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
1081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
1141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
1201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
1261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
1321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
1381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
1441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
1501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
1561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
1621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
1681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
1741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
1801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
1861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
1921 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
1981 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
2041 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
2101 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
2161 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
2221 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
2281 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
2341 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
2401 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
2461 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
2521 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta
2581 atacgactca ctatagggcg aattgggtac gtccctaggc ctccaaaaaa gcctcctcac
2641 tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt
2701 agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga
2761 gttaggggcg ggatagctag agccagacat gataagatac attgatgagt ttggacaaac
2821 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
2881 atttgtaacc attataagct gcaataaaca agttcctctc actctctgat attcatttct
2941 ttgcaagtta taaatactga ataataagat gacatgaact actactgcta gagattttcc
3001 acactgacta aaagggtctg agggatctct agttaccaga gtcacacaac agacgggcac
3061 acactacttg aagcactcaa ggcaagcttt attgaggctt aagcagtggg ttccctagtt
3121 agccagagag ctcccaggct cagatctggt ctaaccagag agacccagta caagcaaaaa
3181 gcagatcttg tcttcgttgg gagtgaatta gcccttccag tccccccttt tcttttaaaa
3241 agtggctaag atctacagct gccttgtaag tcattggtct taaaggtacc atcgccaggc
3301 ggggaggcgg cccaaaggga gatccgactc gtctgagggc gaaggcgaag acgcggaaga
3361 ggccgcagag ccggcagcag gccgcgggaa ggaaggtccg ctggattgag ggccgaaggg
3421 acgtagcaga aggacgtccc gcgcagaatc caggtggcaa cacaggcgag cagccatgga
3481 aaggacgtca gcttccccga caacaccacg gaattgtcag tgcccaacag ccgagcccct
3541 gtccagcagc gggcaaggca ggcggcgatg agttccgccg tggcaatagg gagggggaaa
3601 gcgaaagtcc cggaaaggag ctgacaggtg gtggcaatgc cccaaccagt gggggttgcg
3661 tcagcaaaca cagtgcacac cacgccacgt tgcctgacaa cgggccacaa ctcctcataa
3721 agagacagca accaggattt atacaaggag gagaaaatga aagccatacg ggaagcaata
3781 gcatgataca aaggcattaa agcagcgtat ccacatagcg taaaaggagc aacatagtta
3841 agaataccag tcaatctttc acaaattttg taatccagag gttgattgtc gacgtaccga
3901 tgcatggggt cgtgcgctcc tttcggtcgg gcgctgcggg tcgtggggcg ggcgtcaggc
3961 accgggcttg cgggtcatgc accaggtgcg cggtccttcg ggcacctcga cgtcggcggt
4021 gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg ggcgcggagg tctccaggaa
4081 ggcgggcacc ccggcgcgct cggccgcctc cactccgggg agcacgacgg cgctgcccag
4141 acccttgccc tggtggtcgg gcgagacgcc gacggtggcc aggaaccacg cgggctcctt
4201 gggccggtgc ggcgccagga ggccttccat ctgttgctgc gcggccagcc gggaaccgct
4261 caactcggcc atgcgcgggc cgatctcggc gaacaccgcc cccgcttcga cgctctccgg
4321 cgtggtccag accgccaccg cggcgccgtc gtccgcgacc cacaccttgc cgatgtcgag
4381 cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc cgctcgatgt ggcggtccgg
4441 atcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg gcggcgaggg tgcgtacggc
4501 cctggggacg tcgtcgcggg tggcgaggcg caccgtgggc ttgtactcgg tcatggtaag
4561 ctccggtgga tccccctggg gagagaggtc ggtgattcgg tcaacgaggg agccgactgc
4621 cgacgtgcgc tccggaggct tgcagaatgc ggaacaccgc gcgggcagga acagggccca
4681 cactaccgcc ccacaccccg cctcccgcac cgccccttcc cggccgctgc tctcggcgcg
4741 ccctgctgag cagccgctat tggccacagc ccatcgcggt cggcgcgctg ccattgctcc
4801 ctggcgctgt ccgtctgcga gggtactagt gagacgtgcg gcttccgttt gtcacgtccg
4861 gcacgccgcg aaccgcaagg aaccttcccg acttaggggc ggagcaggaa gcgtcgccgg
4921 ggggcccaca agggtagcgg cgaagatccg ggtgacgctg cgaacggacg tgaagaatgt
4981 gcgagaccca gggtcggcgc cgctgcgttt cccggaacca cgcccagagc agccgcgtcc
5041 ctgcgcaaac ccagggctgc cttggaaaag gcgcaacccc aaccccctcg actaactgac
5101 acacattcca cagaattaat tccaaactca ttactaaccg gtacgcgtag aatcgagacc
5161 gaggagaggg ttagggatag gcttaccttc gaaccgcggg ccctctagac tcgagcggcc
5221 gccactgtgc tggatatcaa ccactttgta caagaaagct gggtcggcgc gcccaccctt
5281 cacggcgatc tttccgccct tcttggcctt tatgaggatc tctctgattt ttcttgcgtc
5341 gagttttccg gtaagacctt tcggtacttc gtccacaaac acaactcctc cgcgcaactt
5401 tttcgcggtt gttacttgac tggcgacgta atccacgatc tctttttccg tcatcgtctt
5461 tccgtgctcc aaaacaacaa cggcggcggg aagttcaccg gcgtcatcgt cgggaagacc
5521 tgcgacacct gcgtcgaaga tgttggggtg ttggagcaag atggattcca attcagcggg
5581 agccacctga tagcctttgt acttaatcag agacttcagg cggtcaacga tgaagaagtg
5641 ttcgtcttcg tcccagtaag ctatgtctcc agaatgtagc catccatcct tgtcaatcaa
5701 ggcgttggtc gcttccggat tgtttacata accggacata atcataggac ctctcacaca
5761 cagttcgcct ctttgattaa cgcccagcgt tttcccggta tccagatcca caaccttcgc
5821 ttcaaaaaat ggaacaactt taccgaccgc gcccggttta tcatccccct cgggtgtaat
5881 cagaatagct gatgtagtct cagtgagccc atatccttgc ctgatacctg gcagatggaa
5941 cctcttggca accgcttccc cgacttcctt agagagggga gcgccaccag aagcaatttc
6001 gtgtaaatta gataaatcgt atttgtcaat cagagtgctt ttggcgaaga aggagaatag
6061 ggttggcacc agcagcgcac tttgaatctt gtaatcctga aggctcctca gaaacagctc
6121 ttcttcaaat ctatacatta agacgactcg aaatccacat atcaaatatc cgagtgtagt
6181 aaacattcca aaaccgtgat ggaatggaac aacacttaaa atcgcagtat ccggaatgat
6241 ttgattgcca aaaataggat ctctggcatg cgagaatctc acgcaggcag ttctatgagg
6301 cagagcgaca cctttaggca gaccagtaga tccagaggag ttcatgatca gtgcaattgt
6361 cttgtcccta tcgaaggact ctggcacaaa atcgtattca ttaaaaccgg gaggtagatg
6421 agatgtgacg aacgtgtaca tcgactgaaa tccctggtaa tccgttttag aatccatgat
6481 aataattttt tggatgattg ggagcttttt ttgcacgttc aaaatttttt gcaacccctt
6541 tttggaaacg aacaccacgg taggctgcga aatgcccata ctgttgagca attcacgttc
6601 attataaatg tcgttcgcgg gcgcaactgc aactccgata aataacgcgc ccaacaccgg
6661 cataaagaat tgaagagagt tttcactgca tacgacgatt ctgtgatttg tattcagccc
6721 atatcgtttc atagcttctg ccaaccgaac ggacatttcg aagtactcag cgtaagtgat
6781 gtccacctcg atatgtgcat ctgtaaaagc aattgttcca ggaaccaggg cgtatctctt
6841 catagcctta tgcagttgct ctccagcggt tccatcttcc agcggataga atggcgccgg
6901 gcctttcttt atgtttttgg cgtcttccat cccggacttg tacagctcgt ccatgccgag
6961 agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg
7021 gtctttgctc agggcggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc
7081 gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat
7141 gttgtggcgg atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata
7201 gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt
7261 gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc
7321 ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc
7381 ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa
7441 gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc
7501 ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc
7561 ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac
7621 cccggtgaac agctcctcgc ccttgctcac catggtgaag ggggcggccg cggagcctgc
7681 ttttttgtac aaacttgttg atatctgcag aattccacca cactggacta gtcgacgatc
7741 tctatcactg atagggagat ctctatcact gatagggaga gctctgctta tatagacctc
7801 ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg
7861 aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact
7921 tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat
7981 caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa
8041 ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg
8101 cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc
8161 cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat
8221 tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta
8281 tgtaagcgga actcccaagc ttatcgataa aattttgaat ttttgtaatt tgtttttgta
8341 attctttagt ttgtatgtct gttgctatta tgtctactat tctttcccct gcactgtacc
8401 ccccaatccc cccttttctt ttaaaagtta accgataccg tcgagatccg ttcactaatc
8461 gaatggatct gtctctgtct ctctctccac cttcttcttc tattccttcg ggcctgtcgg
8521 gtcccctcgg ggttgggagg tgggtctgaa acgataatgg tgaatatccc tgcctaactc
8581 tattcactat agaaagtaca gcaaaaacta ttcttaaacc taccaagcct cctactatca
8641 ttatgaataa ttttatatac cacagccaat ttgttatgtt aaaccaattc cacaaacttg
8701 cccatttatc taattccaat aattcttgtt cattcttttc ttgctggttt tgcgattctt
8761 caattaagga gtgtattaag cttgtgtaat tgttaatttc tctgtcccac tccatccagg
8821 tcgtgtgatt ccaaatctgt tccagagatt tattactcca actagcattc caaggcacag
8881 cagtggtgca aatgagtttt ccagagcaac cccaaatccc caggagctgt tgatccttta
8941 ggtatctttc cacagccagg attcttgcct ggagctgctt gatgccccag actgtgagtt
9001 gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt gttctgctgc tgcactatac
9061 cagacaataa ttgtctggcc tgtaccgtca gcgtcattga cgctgcgccc atagtgcttc
9121 ctgctgctcc caagaaccca aggaacaaag ctcctattcc cactgctctt ttttctctct
9181 gcaccactct tctctttgcc ttggtgggtg ctactcctaa tggttcaatt tttactactt
9241 tatatttata taattcactt ctccaattgt ccctcatatc tcctcctcca ggtctgaaga
9301 tcagcggccg cttgctgtgc ggtggtctta cttttgtttt gctcttcctc tatcttgtct
9361 aaagcttcct tggtgtcttt tatctctatc ctttgatgca cacaatagag ggttgctact
9421 gtattatata atgatctaag ttcttctgat cctgtctgaa gggatggttg tagctgtccc
9481 agtatttgtc tacagccttc tgatgtttct aacaggccag gattaactgc gaatcgttct
9541 agctccctgc ttgcccatac tatatgtttt aatttatatt ttttctttcc ccctggcctt
9601 aaccgaattt tttcccatcg cgatctaatt ctcccccgct taatactgac gctctcgcac
9661 ccatctctct ccttctagcc tccgctagtc aaaatttttg gcgtactcac cagtcgccgc
9721 ccctcgcctc ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga gagctcctct
9781 ggtttccctt tcgctttcaa gtccctgttc gggcgccact gctagagatt ttccacactg
9841 actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta
9901 cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag
9961 agagctccca ggctcagatc tggtctaacc agagagaccc gtttatgtat cgagctaggc
10021 acttaaatac aatatctctg caatgcggca attcagtggt tcgtccaatc catgtcagac
10081 ccgtctgttg ccttcctaat aaggcacgat cgtaccacct tacttccacc aatcggcatg
10141 cacggtgctt tttctctcct tgtaaggcat gttgctaact catcgttacc atgttgcaag
10201 actacaagag tattgcataa gactacatta agcttgcagc tccagctttt gttcccttta
10261 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
10321 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
10381 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
10441 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
//
