| 质粒名称: | pTet-O-Ngn2-puro |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 52047 |
| 存储实验室: | Marius Wernig |
|
载体骨架基本信息 |
|
| 载体名称: | Tet-O-FUW |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 7745 bp |
| 完整质粒大小: | 9861 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，慢病毒表达 |
| 筛选标记: | 嘌呤霉素 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Ngn2 |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 1071 bp |
| 基因突变: | -- |
| GenBank编号 | NM_009718.2 |
| 启动子 | tet-on |
| 标签/融合蛋白: | T2A-and puromycin resistant cassette（插入件上的C端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI(未破坏) |
| 3' 克隆位点: | XbaI (未破坏) |
| 5' 测序引物 | LNCX |
| 3' 测序引物 | WPRE-R |
|
文献引用方法 |
|
| 材料和方法部分: | pTet-O-Ngn2-puro was a gift from Marius Wernig (Addgene plasmid # 52047 ; http://n2t.net/addgene:52047 ; RRID:Addgene_52047) |
| 参考文献部分: | Rapid single-step induction of functional neurons from human pluripotent stem cells. Zhang Y, Pak C, Han Y, Ahlenius H, Zhang Z, Chanda S, Marro S, Patzke C, Acuna C, Covy J, Xu W, Yang N, Danko T, Chen L, Wernig M, Sudhof TC. Neuron. 2013 Jun 5;78(5):785-98. doi: 10.1016/j.neuron.2013.05.029. 10.1016/j.neuron.2013.05.029 PubMed 23764284 |
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 pTet-O-Ngn2-puro 9886 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION 3rd generation lentiviral vector, expresses Ngn2 and puromycin
resistent gene under control of TetON promoter.
ACCESSION .
VERSION .
KEYWORDS pTet-O-Ngn2-puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9886)
AUTHORS Zhang Y, Pak C, Han Y, Ahlenius H, Zhang Z, Chanda S, Marro S,
Patzke C, Acuna C, Covy J, Xu W, Yang N, Danko T, Chen L, Wernig M,
Sudhof TC
TITLE Rapid single-step induction of functional neurons from human
pluripotent stem cells.
JOURNAL Neuron. 2013 Jun 5;78(5):785-98. doi: 10.1016/j.neuron.2013.05.029.
PUBMED 23764284
REFERENCE 2 (bases 1 to 9886)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9886
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 100..479
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 481..679
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 634..654
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 697..877
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 924..1049
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1542..1775
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1960..2004
/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 2302..2419
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 2476..2746
/label=tetracycline response element
/bound_moiety="tetracycline repressor TetR"
/note="contains seven copies of the tetracycline operator
tetO"
promoter 2779..2817
/label=minimal CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2814..2838
/label=LNCX
/note="Human CMV promoter, forward primer"
regulatory 2917..2926
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2926..2949
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 3742..3795
/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 3802..4401
/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(3802..3821)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4298..4318
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 4440..5028
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4493..4513)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4911..4922)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(5031..5047)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5032..5048)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 5553..5733
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(5759..5776)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5765..5989
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 6035..6463
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6122..6141)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6332..6353
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6472..6492)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 6477..6806
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 6657..6792
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 6719..6738
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 6854..6901
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 6920..7294
/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 7424..7545
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7461..7480)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 7515..7534
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(7594..7610)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7594..7610)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7607..7629)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7618..7634
/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(7642..7672)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7687..7708
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7825..7842)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7996..8584)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8076..8095)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(8755..9615)
/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 9378..9397
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9616..9720)
/gene="bla"
/label=AmpR promoter
primer_bind complement(9795..9814)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
61 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
121 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
181 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
241 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
301 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
361 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
421 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
481 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
541 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
601 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
661 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
721 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
781 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
841 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
901 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
961 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
1021 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
1081 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
1141 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
1201 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
1261 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
1321 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
1381 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
1441 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
1501 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
1561 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
1621 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
1681 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
1741 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
1801 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
1861 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
1921 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
1981 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
2041 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
2101 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
2161 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
2221 agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca
2281 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
2341 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
2401 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggttaga
2461 tctcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
2521 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
2581 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
2641 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata
2701 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
2761 gctcggtacc cgggtcgagg taggcgtgta cggtgggagg cctatataag cagagctcgt
2821 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
2881 caccgggacc gatccagcct ccgcggcccc gaattcgcca ccatggacta caaggacgac
2941 gatgacaagt tcgtcaaatc tgagactctg gagttgaagg aggaagagga ggtactgatg
3001 ctgctgggct cggcttcccc ggcctcggcg accctgaccc cgatgtcctc cagcgcggac
3061 gaggaggagg acgaggagct gcgccggccg ggctccgcgc gtgggcagcg tggagcggaa
3121 gccgggcagg gggtgcaggg cagtccggcg tcgggtgccg ggggttgccg gccagggcgg
3181 ctgctgggcc tgatgcacga gtgcaagcgt cgcccgtcgc gctcacgggc cgtctcccga
3241 ggtgccaaga cggcggagac ggtgcagcgc atcaagaaga cccgcaggct caaggccaac
3301 aaccgcgagc gcaaccgcat gcacaaccta aacgccgcgc tggacgcgct gcgcgaggtg
3361 ctgcccacct tccccgagga tgccaagctc acgaagatcg agacgctgcg cttcgcccac
3421 aattacatct gggcgctcac cgagactctg cgcctggcgg accactgcgc cggcgccggt
3481 ggcctccagg gggcgctctt cacggaggcg gtgctcctga gcccgggagc tgcgctcggc
3541 gccagcgggg acagcccttc tccaccttcc tcctggagct gcaccaacag cccggcgtca
3601 tcctccaact ccacgtcccc atacagctgc actttatcgc ccgctagccc cgggtcagac
3661 gtggactact ggcagccccc acctccggag aagcatcgtt atgcgcctca cctgcccctc
3721 gccagggact gtatctctag agagggcagg ggaagtcttc taacatgcgg ggacgtggag
3781 gaaaatcccg gcccactcga gatgaccgag tacaagccca cggtgcgcct cgccacccgc
3841 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
3901 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
3961 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
4021 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
4081 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
4141 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
4201 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
4261 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
4321 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
4381 acccgcaagc ccggtgcctg attaattaag ctagctaatt cgatatcaag cttatcgata
4441 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
4501 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
4561 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
4621 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
4681 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
4741 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
4801 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
4861 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
4921 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
4981 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
5041 gacctcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg
5101 ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc
5161 aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
5221 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt
5281 ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca gggccaggga
5341 tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag caagagaagg
5401 tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc ctgcatggga
5461 tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta gcatttcatc
5521 acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac cagatctgag
5581 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
5641 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
5701 gaccctttta gtcagtgtgg aaaatctcta gcagggcccg tttaaacccg ctgatcagcc
5761 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
5821 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
5881 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
5941 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
6001 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
6061 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
6121 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
6181 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
6241 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
6301 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
6361 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
6421 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
6481 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
6541 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
6601 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
6661 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
6721 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
6781 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
6841 ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
6901 caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg
6961 cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt
7021 ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca
7081 ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct
7141 gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat
7201 gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa
7261 ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt tcgattccac
7321 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
7381 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
7441 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
7501 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
7561 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
7621 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
7681 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
7741 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
7801 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
7861 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
7921 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
7981 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
8041 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
8101 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
8161 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
8221 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
8281 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
8341 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
8401 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
8461 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
8521 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
8581 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
8641 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
8701 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
8761 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
8821 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
8881 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
8941 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
9001 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
9061 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
9121 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
9181 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
9241 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
9301 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
9361 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
9421 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
9481 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
9541 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
9601 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
9661 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
9721 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
9781 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
9841 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaag
//
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