| 质粒名称: | pLV-EF1A-CD19.28H.28TM.28.3z |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 200678 |
| 存储实验室: | Michele Bernasconi |
|
载体骨架基本信息 |
|
| 载体名称: | pLV |
| 载体出品公司: | VectorBuilder |
| 空载体大小: | 6451 bp |
| 完整质粒大小: | 8753bp |
| 载体类型: | 哺乳细胞表达, Lentiviral |
| 筛选标记: | 新霉素(G418) |
| 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: | FMC63-CD28HD-CD28TM-CD28-CD3z-P2A-EGFP、CD19(FMC63)-CAR, CD28 hinge, CD28 、Contains: FMC63-scFV, CD28 Hinge domain, CD28-transmembrane domain, CD28-ITD, CD3z-ITD |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 2334bp |
| 启动子: | EF1a-short |
| 标签/融合蛋白: | MycTag (N terminal on insert) |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | tgatgtcgtgtactggctcc |
| 3’测序引物 | tttcttccacgtcgcctgcttg |
|
文献引用方法 |
|
| 材料和方法部分: | pLV-EF1A-CD19.28H.28TM.28.3z was a gift from Michele Bernasconi (Addgene plasmid # 200678 ; http://n2t.net/addgene:200678 ; RRID:Addgene_200678) |
| 参考文献部分: | CD276-CAR T cells and Dual-CAR T cells targeting CD276/FGFR4 promote rhabdomyosarcoma clearance in orthotopic mouse models. Timpanaro A, Piccand C, Dzhumashev D, Anton-Joseph S, Robbi A, Moser J, Rossler J, Bernasconi M. J Exp Clin Cancer Res. 2023 Nov 4;42(1):293. doi: 10.1186/s13046-023-02838-3. 10.1186/s13046-023-02838-3 PubMed 37924157 |
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pLV-EF1A-CD19.28H.28TM.28.3z载体质粒基本信息
pLV-EF1A-CD19.28H.28TM.28.3z质粒图谱载体图谱和 pLV-EF1A-CD19.28H.28TM.28.3z载体序列质粒序列多克隆位点信息

pLV-EF1A-CD19.28H.28TM.28.3z质粒载体简介
pLV-EF1A-CD19.28H.28TM.28.3z质粒序列载体序列
LOCUS pLV-EF1A-CD19.28H.28TM.28.3z 8753 bp DNA circular SYN 14-APR-2025
DEFINITION Modular CD28hinge-CD28transmembrane-CD28-CD3z CAR backbone, For
Transient Expression or Lentiviral Production.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8753)
AUTHORS Timpanaro A, Piccand C, Dzhumashev D, Anton-Joseph S, Robbi A, Moser
J, Rossler J, Bernasconi M
TITLE CD276-CAR T cells and Dual-CAR T cells targeting CD276/FGFR4 promote
rhabdomyosarcoma clearance in orthotopic mouse models.
JOURNAL J Exp Clin Cancer Res. 2023 Nov 4;42(1):293. doi:
10.1186/s13046-023-02838-3.
PUBMED 37924157
REFERENCE 2 (bases 1 to 8753)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Exp Clin
Cancer Res."; date: "2023-11-4"; volume: "42(1):293. doi"; pages: "
10.1186/s13046-023-02838-3"
COMMENT Sequence Label: pLV-EF1A-CD19.28H.28TM.28.3z
FEATURES Location/Qualifiers
source 1..8753
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 3..229
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 230..410
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 457..582
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1075..1308
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1493..1537
/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 1803..1920
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 1938..1958
/gene="mutant version of attB"
/label=attB4
/bound_moiety="BP Clonase(TM)"
/note="core recombination site for the Gateway(R) BP
reaction"
promoter 1965..2176
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
protein_bind 2182..2206
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 2273..2302
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2303..2332
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 3773..3829
/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 join(3830..3832,3833..4546)
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
CDS join(3830..3832,3833..4543)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(3872..3893)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4133..4152)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4480..4501
/label=EGFP-C
/note="EGFP, forward primer"
protein_bind complement(4556..4580)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
misc_feature 4598..5186
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4651..4671)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5069..5080)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 5274..5507
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 5579..5700
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5616..5635)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5670..5689
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 5740..5875
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(5895..5914)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(5896..5914)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5924..5941)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(5924..5940)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5933..5955)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 6082..6537
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6169..6188)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6379..6400
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 6563..6667
/gene="bla"
/label=AmpR promoter
CDS join(6668..6736,6737..7528)
/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(6886..6905)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7699..8287
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8188..8207
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8441..8458
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 8575..8596
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter join(8611..8616,8617..8634,8635..8641)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8649..8665
/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 8654..8676
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 8673..8689
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 8673..8689
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 8708..8728
/label=T3
/note="T3 promoter, forward primer"
promoter 8710..8728
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgcta
1801 gcttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
1861 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt
1921 actagtgatt atcggatcaa ctttgtatag aaaagttgtc tagagggcag agcgcacatc
1981 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
2041 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
2101 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
2161 tgccgccaga acacagggat cccaagtttg tacaaaaaag caggctgcca ccatgctcag
2221 gctgctcttg gctctcaact tattcccttc aattcaagta acaggactcg aggaacaaaa
2281 actcatctca gaagaggatc tggaacaaaa gctcatttca gaagaagatc tgctcgaggt
2341 cgacggagac gccgacatcc agatgacaca gactacatcc tccctgtctg cctctctggg
2401 agacagagtc accatcagtt gcagggcaag tcaggacatc tctaagtatt tgaattggta
2461 tcagcagaaa ccagatggaa ctgttaaact cctgatctac catacatcaa gattacactc
2521 aggagtccca tcaaggttca gtggcagtgg gtctggaaca gattattctc tcaccattag
2581 caacctggag caagaagata ttgccactta cttttgccaa cagggtaata cgcttccgta
2641 cacgttcgga ggggggacta agttggaaat aacaggctcc acctctggat ccggcaagcc
2701 cggatctggc gagggatcca ccaagggcga ggtgaaactg caggagtcag gacctggcct
2761 ggtggcgccc tcacagagcc tgtccgtcac atgcactgtc tcaggggtct cattacccga
2821 ctatggtgta agctggattc gccagcctcc acgaaagggt ctggagtggc tgggagtaat
2881 atggggtagt gaaaccacat actataattc agctctcaaa tccagactga ccatcatcaa
2941 ggacaactcc aagagccaag ttttcttaaa aatgaacagt ctgcaaactg atgacacagc
3001 catttactac tgtgccaaac attattacta cggtggtagc tatgctatgg actactgggg
3061 tcaaggaacc tcagtcaccg tctccggtac cattgaagtt atgtatcctc ctccttacct
3121 agacaatgag aagagcaatg gaaccattat ccatgtgaaa gggaaacacc tttgtccaag
3181 tcccctattt cccggacctt ctaagccctt ttgggtgctg gtggtggttg gtggagtcct
3241 ggcttgctat agcttgctag taacagtggc ctttattatt ttctgggtga ggagtaagag
3301 gagcaggctc ctgcacagtg actacatgaa catgactccc aggcggcccg gacccacccg
3361 caagcattac cagccctatg ccccaccacg cgacttcgca gcctatcgct cctccggact
3421 gagagtgaag ttcagcagga gcgcagacgc ccccgcgtac cagcagggcc agaaccagct
3481 ctataacgag ctcaatctag gacgaagaga ggagtacgat gttttggaca agaggcgtgg
3541 ccgggaccct gagatggggg gaaagccgca gagaaggaag aaccctcagg aaggcctgta
3601 caatgaactg cagaaagata agatggcgga ggcctacagt gagattggga tgaaaggcga
3661 gcgccggagg ggcaaggggc acgatggcct ttaccagggt ctcagtacag ccaccaagga
3721 cacctacgac gcccttcaca tgcaggccct gccccctcgc tccggacgta cggccacgaa
3781 cttctctctg ttaaagcaag caggcgacgt ggaagaaaac cccggtcccg tgagcaaggg
3841 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
3901 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
3961 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
4021 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
4081 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
4141 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
4201 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
4261 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
4321 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
4381 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
4441 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
4501 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagggc cccgtaccca
4561 gctttcttgt acaaagtggt gataatcgaa ttccgataat caacctctgg attacaaaat
4621 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
4681 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
4741 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
4801 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
4861 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
4921 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
4981 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
5041 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
5101 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
5161 gatctccctt tgggccgcct ccccgcatcg ggaattcccg cggttcgctt taagaccaat
5221 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
5281 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt
5341 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
5401 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
5461 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
5521 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
5581 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
5641 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
5701 tcatgtctgg ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc
5761 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
5821 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg
5881 acgtacccaa ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac
5941 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
6001 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
6061 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
6121 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
6181 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
6241 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
6301 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
6361 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
6421 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
6481 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
6541 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
6601 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
6661 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
6721 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
6781 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
6841 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
6901 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
6961 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
7021 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
7081 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
7141 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
7201 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
7261 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
7321 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
7381 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
7441 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
7501 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
7561 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
7621 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
7681 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
7741 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
7801 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
7861 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
7921 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
7981 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
8041 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
8101 cgccacgctt cccgaagaga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
8161 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
8221 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
8281 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
8341 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
8401 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
8461 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
8521 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
8581 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
8641 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
8701 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag ctt
//
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