| 质粒名称: | pLenti_PE7-2A-BSD |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 234073 |
| 存储实验室: | Greg Findlay |
|
载体骨架基本信息 |
|
| 载体名称: | pLenti-PE2-BSD |
| 载体出品公司: | Hyongbum Kim, Addgene plasmid #161514 |
| 空载体大小: | 8693bp |
| 完整质粒大小: | 15764bp |
| 载体类型: | 哺乳细胞表达, Lentiviral, CRISPR |
| 筛选标记: | Blasticidin |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 30 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | PE7 |
| 物种来源: | Synthetic; SpCas9 is from Streptococcus pyogenes; MMLV-RT is from the Moloney murine leukemia virus; La is from Homo sapiens |
| 插入大小: | 7071bp |
| GenBank编号 | JN247409 |
| 启动子: | EF-1α core promoter |
| 标签/融合蛋白: |
SV40 bpNLS (N terminal on insert)
SV40 bpNLS (C terminal on insert)
c-Myc NLS (C terminal on insert)
|
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | EFS_forward (5'-GGTCGGCAATTGATCCGGTG) |
|
文献引用方法 |
|
| 材料和方法部分: | pLenti_PE7-2A-BSD was a gift from Greg Findlay (Addgene plasmid # 234073 ; http://n2t.net/addgene:234073 ; RRID:Addgene_234073) |
| 参考文献部分: | High-throughput screening of human genetic variants by pooled prime editing. Herger M, Kajba CM, Buckley M, Cunha A, Strom M, Findlay GM. Cell Genom. 2025 Mar 18:100814. doi: 10.1016/j.xgen.2025.100814. 10.1016/j.xgen.2025.100814 PubMed 40120586 |
买家导航
pLenti_PE7-2A-BSD载体质粒基本信息
pLenti_PE7-2A-BSD质粒图谱载体图谱和pLenti_PE7-2A-BSD载体序列质粒序列多克隆位点信息
pLenti_PE7-2A-BSD质粒载体简介
pLenti_PE7-2A-BSD质粒序列载体序列
LOCUS pLenti_PE7-2A-BSD 15764 bp DNA circular SYN 15-APR-2025
DEFINITION For lentiviral expression of PE7 with blasticidin resistance.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 15764)
AUTHORS Herger M, Kajba CM, Buckley M, Cunha A, Strom M, Findlay GM
TITLE High-throughput screening of human genetic variants by pooled prime
editing.
JOURNAL Cell Genom. 2025 Mar 18:100814. doi: 10.1016/j.xgen.2025.100814.
PUBMED 40120586
REFERENCE 2 (bases 1 to 15764)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 15, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell Genom.
2025 Mar 18:100814. doi: 10.1016/j.xgen.2025.100814."
COMMENT Sequence Label: pLenti_PE7-2A-BSD
FEATURES Location/Qualifiers
source 1..15764
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 7..386
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 388..586
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 541..561
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 604..784
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 831..956
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1449..1682
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1867..1911
/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 2209..2326
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2419..2630
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
CDS 2686..2706
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2707..6807
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRKLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLKREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 6865..6885
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 9664..9684
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 9694..9720
/codon_start=1
/product="nuclear localization signal of human c-Myc
proto-oncogene (Dang and Lee, 1988)"
/label=c-myc NLS
/translation="PAAKRVKLD"
CDS 9730..9786
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 9787..10185
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
misc_feature 10246..10834
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(10299..10319)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(10717..10728)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(10837..10853)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(10838..10854)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 11359..11539
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(11565..11582)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 11571..11795
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 11841..12269
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(11928..11947)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 12138..12159
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(12278..12298)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 12443..12577
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 12504..12523
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 12639..12686
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 12705..13079
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 13209..13330
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(13246..13265)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 13300..13319
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(13379..13395)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(13379..13395)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(13392..13414)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 13403..13419
/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(join(13427..13433,13434..13451,13452..13457))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 13472..13493
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(13610..13627)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(13781..14369)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(13861..13880)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(14540..15331,15332..15400))
/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 15163..15182
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(15401..15505)
/gene="bla"
/label=AmpR promoter
primer_bind complement(15580..15599)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
601 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
661 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
721 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
781 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
841 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
901 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
961 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
1021 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
1081 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
1141 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
1201 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
1261 agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga
1321 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
1381 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
1441 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
1501 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
1561 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
1621 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
1681 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
1741 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
1801 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
1861 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
1921 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
1981 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
2041 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
2101 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg
2161 gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa
2221 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
2281 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
2341 ggacagcaga gatccagttt ggttaattag ctagctaggt cttgaaagga gtgggaattg
2401 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
2461 gaggggtcgg caattgatcc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
2521 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
2581 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gaccggttct
2641 agagccacca tgaaacggac agccgacgga agcgagttcg agtcaccaaa gaagaagcgg
2701 aaagtcgaca agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc
2761 gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac
2821 cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc
2881 gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc
2941 tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac
3001 agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc
3061 ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga
3121 aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc
3181 cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc
3241 gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac
3301 cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc
3361 agaaagctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga
3421 aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc
3481 gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg
3541 gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc
3601 atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc
3661 tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg
3721 cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac
3781 gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc
3841 ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaagagaga ggacctgctg
3901 cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg
3961 cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag
4021 atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac
4081 agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag
4141 gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat
4201 aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc
4261 gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc
4321 ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg
4381 accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa
4441 atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa
4501 attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc
4561 gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat
4621 gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg
4681 ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc
4741 ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac
4801 gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc
4861 ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag
4921 acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc
4981 gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag
5041 agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac
5101 cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg
5161 cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac
5221 gctatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga
5281 agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg
5341 aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat
5401 ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga
5461 cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg
5521 aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag
5581 tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac
5641 aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa
5701 aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg
5761 aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac
5821 agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag
5881 cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat
5941 tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag
6001 gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg
6061 atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg
6121 gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt
6181 gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc
6241 gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct
6301 aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa
6361 ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc
6421 agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg
6481 gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga
6541 gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat
6601 aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga
6661 gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc
6721 aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg
6781 atcgacctgt ctcagctggg aggtgactcc ggcggaagct ctggtggcag caagcggacc
6841 gccgacggct ctgaattcga gagccctaag aagaaaagaa aggtgagcgg aggctctagc
6901 ggcggaagca ccctgaacat tgaagacgag tatagactgc atgaaacaag caaggaaccc
6961 gatgtgtccc tgggctccac ctggctgtcc gactttcccc aggcctgggc cgagacagga
7021 ggaatgggcc tggccgtgcg gcaggcaccc ctgatcatcc ctctgaaggc cacctctaca
7081 cccgtgagca tcaagcagta ccctatgtct caggaggcca gactgggcat caagcctcac
7141 atccagaggc tgctggacca gggcatcctg gtgccatgcc agagcccctg gaacacacca
7201 ctgctgcccg tgaagaagcc aggcaccaat gactatagac ccgtgcagga tctgagagag
7261 gtgaacaaga gggtggagga tatccacccc accgtgccca acccttacaa tctgctgtcc
7321 ggcctgcccc cttctcacca gtggtataca gtgctggacc tgaaggatgc cttcttttgt
7381 ctgagactgc accctaccag ccagccactg ttcgcctttg agtggaggga ccctgagatg
7441 ggcatctctg gccagctgac ctggacacgc ctgcctcagg gcttcaagaa tagcccaaca
7501 ctgtttaacg aggccctgca ccgcgacctg gcagatttcc ggatccagca cccagatctg
7561 atcctgctgc agtacgtgga cgatctgctg ctggccgcca ccagcgagct ggattgccag
7621 cagggaacac gcgccctgct gcagaccctg ggaaacctgg gatatagggc atccgccaag
7681 aaggcccaga tctgtcagaa gcaggtgaag tacctgggct atctgctgaa ggagggccag
7741 agatggctga cagaggccag gaaggagaca gtgatgggcc agccaacacc caagacccca
7801 agacagctga gggagttcct gggcaaagca ggattttgca ggctgttcat cccaggattc
7861 gcagagatgg cagcacctct gtacccactg accaagccgg gcaccctgtt taattggggc
7921 cctgaccagc agaaggccta tcaggagatc aagcaggccc tgctgacagc accagccctg
7981 ggcctgccag acctgaccaa gcctttcgag ctgtttgtgg atgagaagca gggctacgcc
8041 aagggcgtgc tgacccagaa gctgggacca tggagacggc ccgtggccta tctgtccaag
8101 aagctggacc cagtggcagc aggatggcca ccatgcctga ggatggtggc agcaatcgcc
8161 gtgctgacaa aggatgccgg caagctgacc atgggacagc cactggtcat cctggcacca
8221 cacgcagtgg aggccctggt gaagcagcct ccagatcgct ggctgtctaa cgcccggatg
8281 acacactacc aggccctgct gctggacacc gatcgcgtgc agtttggccc tgtggtggcc
8341 ctgaatccag ccaccctgct gcctctgcca gaggagggcc tgcagcacaa ctgtctggac
8401 atcctggcag aggcacacgg aacaaggcca gacctgaccg atcagcccct gcctgacgcc
8461 gatcacacat ggtataccga tggaagctcc ctgctgcagg agggccagag gaaggcagga
8521 gcagcagtga ccacagagac agaagtgatc tgggccaagg ccctgccagc aggcacatcc
8581 gcccagcggg ccgagctgat cgccctgacc caggccctga agatggccga gggcaagaag
8641 ctgaacgtgt acacagactc cagatatgcc ttcgccaccg cacacatcca cggagagatc
8701 tacaggcgcc ggggctggct gacctctgag ggcaaggaga tcaagaacaa ggatgagatc
8761 ctggccctgc tgaaggccct gtttctgccc aagcggctga gcatcatcca ctgtcctgga
8821 caccagaagg gacactccgc cgaggcaagg ggcaatcgga tggccgacca ggccgccaga
8881 aaggctgcta ttactgaaac tcccgacact tccactctgc tgattgaaaa ctcctcccct
8941 tccggaggat ctagcggagg ctcctctggc tctgagacac ctggcacaag cgagagcgca
9001 acacctgaaa gcagcggggg cagcagcggg gggtcaatgg ctgaaaatgg tgataatgaa
9061 aagatggctg ccctggaggc caaaatctgt catcaaattg agtattattt tggcgacttc
9121 aatttgccac gggacaagtt tctaaaggaa cagataaaac tggatgaagg ctgggtacct
9181 ttggagataa tgataaaatt caacaggttg aaccgtctaa caacagactt taatgtaatt
9241 gtggaagcat tgagcaaatc caaggcagaa ctcatggaaa tcagtgaaga taaaactaaa
9301 atcagaaggt ctccaagcaa acccctacct gaagtgactg atgagtataa aaatgatgta
9361 aaaaacagat ctgtttatat taaaggcttc ccaactgatg caactcttga tgacataaaa
9421 gaatggttag aagataaagg tcaagtacta aatattcaga tgagaagaac attgcataaa
9481 gcatttaagg gatcaatttt tgttgtgttt gatagcattg aatctgctaa gaaatttgta
9541 gagacccctg gccagaagta caaagaaaca gacctgctaa tacttttcaa ggacgattac
9601 tttgccaaaa aaaatgaatc tggcggctca aaaagaaccg ccgacggcag cgaattcgag
9661 tctcccaaga agaagaggaa agtcggctct ggccctgccg ctaagagagt gaagctggac
9721 ggatccggcg caacaaactt ctctctgctg aaacaagccg gagatgtcga agagaatcct
9781 ggaccgatgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct
9841 acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc tctctctagc
9901 gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa
9961 ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg
10021 atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc
10081 gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt
10141 gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaagccgt gacctacatg
10201 ctcgaagggc gtatgcgcca cgaattcgat atcaagctta tcggtaatca acctctggat
10261 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
10321 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
10381 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
10441 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
10501 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
10561 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
10621 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
10681 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
10741 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
10801 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgagaccta
10861 gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg
10921 ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga
10981 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
11041 gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca
11101 caaggctact tccctgattg gcagaactac acaccagggc cagggatcag atatccactg
11161 acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga agaagccaat
11221 gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga tgacccggag
11281 agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat ggcccgagag
11341 ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
11401 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
11461 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
11521 gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
11581 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
11641 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
11701 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
11761 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
11821 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
11881 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
11941 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
12001 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
12061 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
12121 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
12181 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
12241 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
12301 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
12361 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
12421 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccagttccgc ccattctccg
12481 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
12541 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
12601 ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat
12661 agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc aagttgacca
12721 gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc
12781 ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg
12841 tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg
12901 tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact
12961 tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt
13021 tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac
13081 acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
13141 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
13201 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
13261 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
13321 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
13381 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
13441 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
13501 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
13561 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
13621 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
13681 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
13741 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
13801 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
13861 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
13921 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
13981 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
14041 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
14101 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
14161 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
14221 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
14281 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
14341 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
14401 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
14461 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
14521 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
14581 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
14641 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
14701 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
14761 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
14821 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
14881 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
14941 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
15001 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
15061 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
15121 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
15181 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
15241 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
15301 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
15361 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
15421 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
15481 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
15541 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
15601 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
15661 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
15721 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tata
//
