| 质粒名称: | pdCASclos |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 73640 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pIMP1-ptb |
| 完整质粒大小: | 9056 bp |
| 载体类型: | E.coli-clostridium shuttle vector |
| 筛选标记: |
Erythromycin in clostridium
GROWTH IN BACTERIA
|
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称1: | dCas9 |
| 物种来源: | S. pygogenes |
| 基因突变: | D10A, H840A |
| 启动子: | ptb |
| 克隆方法: | 未知 |
| 5’测序引物 | SpCas9N-R TACGAGCTGTCCGTTTGAGA |
| 插入基因名称2: | sgRNA to spo0A |
| 启动子: | Pj23119 |
| 克隆方法: | 未知 |
| 5’测序引物 | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | pdCASclos was a gift from Sheng Yang (Addgene plasmid # 73640 ; http://n2t.net/addgene:73640 ; RRID:Addgene_73640) |
| 参考文献部分: | CRISPR-based genome editing and expression control systems in Clostridium acetobutylicum and Clostridium beijerinckii. Li Q, Chen J, Minton NP, Zhang Y, Wen Z, Liu J, Yang H, Zeng Z, Ren X, Yang J, Gu Y, Jiang W, Jiang Y, Yang S. Biotechnol J. 2016 May 23. doi: 10.1002/biot.201600053. 10.1002/biot.201600053 PubMed 27213844 |
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 pdCASclos 9056 bp ds-DNA
circular SYN 12-MAY-2021
DEFINITION Transcriptional repression for gene Spo0A
(CAC-2011) in Clostridium
acetobutylicum ATCC 824.
ACCESSION .
VERSION .
KEYWORDS pdCASclos
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9056)
AUTHORS Li Q, Chen J, Minton NP, Zhang Y, Wen Z, Liu
J, Yang H, Zeng Z, Ren
X, Yang J, Gu Y, Jiang W, Jiang Y, Yang S
TITLE CRISPR-based genome editing and expression
control systems in
Clostridium acetobutylicum and Clostridium
beijerinckii.
JOURNAL Biotechnol J. 2016 May 23. doi:
10.1002/biot.201600053.
PUBMED 27213844
REFERENCE 2 (bases 1 to 9056)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server
1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9056
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 14..48
/label=J23119(SpeI) promoter
/note="bacterial promoter (Registry
of Standard Biological
Parts BBa_J23119) modified to end
with an SpeI site"
misc_RNA 69..144
/label=gRNA scaffold
/note="guide RNA scaffold for the
Streptococcus pyogenes
CRISPR/Cas9 system"
CDS 281..4387
/codon_start=1
/product="catalytically dead mutant
of the Cas9
endonuclease from the Streptococcus
pyogenes Type II
CRISPR/Cas system"
/label=dCas9
/note="RNA-guided DNA-binding
protein that lacks
endonuclease activity due to the
D10A mutation in the RuvC
catalytic domain and the H840A
mutation in the HNH
catalytic domain"
/translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSI
KK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLE
ES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHM
IK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSR
RL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLL
AQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKAL
VR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNRED
LL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLA
RG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLY
EY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIEC
FD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIE
ER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRN
FM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVM
GR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEK
LY
LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDN
VP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQIT
KH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDA
YL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFF
KT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGF
SK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELL
GI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKG
NE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVIL
AD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKE
VL
DATLIHQSITGLYETRIDLSQLGGD"
primer_bind complement(4397..4414)
/label=M13 Forward
/note="In lacZ gene. Also called
M13-F20 or M13 (-21)
Forward"
primer_bind complement(4397..4413)
/label=M13 fwd
/note="common sequencing primer, one
of multiple similar
variants"
primer_bind complement(4406..4428)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(4622..4641)
/label=pRS-marker
/note="pRS vectors, use to sequence
yeast selectable
marker"
primer_bind 4741..4763
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4801..4819)
/label=pBRforEco
/note="pBR322 vectors, upsteam of
EcoRI site, forward
primer"
promoter 4887..4991
/gene="bla"
/label=AmpR promoter
CDS 4992..5852
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to
ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVG
YI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLD
RW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLR
SA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIG
AS
LIKHW"
primer_bind complement(5210..5229)
/label=Amp-R
/note="Ampicillin resistance gene,
reverse primer"
rep_origin 6023..6611
/direction=RIGHT
/label=ori
/note="high-copy-number
ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6512..6531
/label=pBR322ori-F
/note="pBR322 origin, forward
primer"
primer_bind 6765..6782
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 6899..6920
/label=CAP binding site
/bound_moiety="E. coli catabolite
activator protein"
/note="CAP binding activates
transcription in the presence
of cAMP."
promoter 6935..6965
/label=lac promoter
/note="promoter for the E. coli lac
operon"
protein_bind 6973..6989
/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 6978..7000
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 6997..7013
/label=M13 rev
/note="common sequencing primer, one
of multiple similar
variants"
primer_bind 6997..7013
/label=M13 Reverse
/note="In lacZ gene. Also called
M13-rev"
ORIGIN
1 agcttggctg cagttgacag ctagctcagt cctaggtata
atactagttt taattgcaga
61 tgataatagt tttagagcta gaaatagcaa gttaaaataa
ggctagtccg ttatcaactt
121 gaaaaagtgg caccgagtcg gtgctttttt gcggccgcta
taaaatataa ataattttct
181 aaaaaactta acttcatgtg aaaagtttgt taaaatataa
atgagcacgt taatcattta
241 acatagataa ttaaatagta aaagggagtg tcgactcgag
atggataaga aatactcaat
301 aggcttagct atcggcacaa atagcgtcgg atgggcggtg
atcactgatg aatataaggt
361 tccgtctaaa aagttcaagg ttctgggaaa tacagaccgc
cacagtatca aaaaaaatct
421 tataggggct cttttatttg acagtggaga gacagcggaa
gcgactcgtc tcaaacggac
481 agctcgtaga aggtatacac gtcggaagaa tcgtatttgt
tatctacagg agattttttc
541 aaatgagatg gcgaaagtag atgatagttt ctttcatcga
cttgaagagt cttttttggt
601 ggaagaagac aagaagcatg aacgtcatcc tatttttgga
aatatagtag atgaagttgc
661 ttatcatgag aaatatccaa ctatctatca tctgcgaaaa
aaattggtag attctactga
721 taaagcggat ttgcgcttaa tctatttggc cttagcgcat
atgattaagt ttcgtggtca
781 ttttttgatt gagggagatt taaatcctga taatagtgat
gtggacaaac tatttatcca
841 gttggtacaa acctacaatc aattatttga agaaaaccct
attaacgcaa gtggagtaga
901 tgctaaagcg attctttctg cacgattgag taaatcaaga
cgattagaaa atctcattgc
961 tcagctcccc ggtgagaaga aaaatggctt atttgggaat
ctcattgctt tgtcattggg
1021 tttgacccct aattttaaat caaattttga tttggcagaa
gatgctaaat tacagctttc
1081 aaaagatact tacgatgatg atttagataa tttattggcg
caaattggag atcaatatgc
1141 tgatttgttt ttggcagcta agaatttatc agatgctatt
ttactttcag atatcctaag
1201 agtaaatact gaaataacta aggctcccct atcagcttca
atgattaaac gctacgatga
1261 acatcatcaa gacttgactc ttttaaaagc tttagttcga
caacaacttc cagaaaagta
1321 taaagaaatc ttttttgatc aatcaaaaaa cggatatgca
ggttatattg atgggggagc
1381 tagccaagaa gaattttata aatttatcaa accaatttta
gaaaaaatgg atggtactga
1441 ggaattattg gtgaaactaa atcgtgaaga tttgctgcgc
aagcaacgga cctttgacaa
1501 cggctctatt ccccatcaaa ttcacttggg tgagctgcat
gctattttga gaagacaaga
1561 agacttttat ccatttttaa aagacaatcg tgagaagatt
gaaaaaatct tgacttttcg
1621 aattccttat tatgttggtc cattggcgcg tggcaatagt
cgttttgcat ggatgactcg
1681 gaagtctgaa gaaacaatta ccccatggaa ttttgaagaa
gttgtcgata aaggtgcttc
1741 agctcaatca tttattgaac gcatgacaaa ctttgataaa
aatcttccaa atgaaaaagt
1801 actaccaaaa catagtttgc tttatgagta ttttacggtt
tataacgaat tgacaaaggt
1861 caaatatgtt actgaaggaa tgcgaaaacc agcatttctt
tcaggtgaac agaagaaagc
1921 cattgttgat ttactcttca aaacaaatcg aaaagtaacc
gttaagcaat taaaagaaga
1981 ttatttcaaa aaaatagaat gttttgatag tgttgaaatt
tcaggagttg aagatagatt
2041 taatgcttca ttaggtacct accatgattt gctaaaaatt
attaaagata aagatttttt
2101 ggataatgaa gaaaatgaag atatcttaga ggatattgtt
ttaacattga ccttatttga
2161 agatagggag atgattgagg aaagacttaa aacatatgct
cacctctttg atgataaggt
2221 gatgaaacag cttaaacgtc gccgttatac tggttgggga
cgtttgtctc gaaaattgat
2281 taatggtatt agggataagc aatctggcaa aacaatatta
gattttttga aatcagatgg
2341 ttttgccaat cgcaatttta tgcagctgat ccatgatgat
agtttgacat ttaaagaaga
2401 cattcaaaaa gcacaagtgt ctggacaagg cgatagttta
catgaacata ttgcaaattt
2461 agctggtagc cctgctatta aaaaaggtat tttacagact
gtaaaagttg ttgatgaatt
2521 ggtcaaagta atggggcggc ataagccaga aaatatcgtt
attgaaatgg cacgtgaaaa
2581 tcagacaact caaaagggcc agaaaaattc gcgagagcgt
atgaaacgaa tcgaagaagg
2641 tatcaaagaa ttaggaagtc agattcttaa agagcatcct
gttgaaaata ctcaattgca
2701 aaatgaaaag ctctatctct attatctcca aaatggaaga
gacatgtatg tggaccaaga
2761 attagatatt aatcgtttaa gtgattatga tgtcgatgca
attgttccac aaagtttcct
2821 taaagacgat tcaatagaca ataaggtctt aacgcgttct
gataaaaatc gtggtaaatc
2881 ggataacgtt ccaagtgaag aagtagtcaa aaagatgaaa
aactattgga gacaacttct
2941 aaacgccaag ttaatcactc aacgtaagtt tgataattta
acgaaagctg aacgtggagg
3001 tttgagtgaa cttgataaag ctggttttat caaacgccaa
ttggttgaaa ctcgccaaat
3061 cactaagcat gtggcacaaa ttttggatag tcgcatgaat
actaaatacg atgaaaatga
3121 taaacttatt cgagaggtta aagtgattac cttaaaatct
aaattagttt ctgacttccg
3181 aaaagatttc caattctata aagtacgtga gattaacaat
taccatcatg cccatgatgc
3241 gtatctaaat gccgtcgttg gaactgcttt gattaagaaa
tatccaaaac ttgaatcgga
3301 gtttgtctat ggtgattata aagtttatga tgttcgtaaa
atgattgcta agtctgagca
3361 agaaataggc aaagcaaccg caaaatattt cttttactct
aatatcatga acttcttcaa
3421 aacagaaatt acacttgcaa atggagagat tcgcaaacgc
cctctaatcg aaactaatgg
3481 ggaaactgga gaaattgtct gggataaagg gcgagatttt
gccacagtgc gcaaagtatt
3541 gtccatgccc caagtcaata ttgtcaagaa aacagaagta
cagacaggcg gattctccaa
3601 ggagtcaatt ttaccaaaaa gaaattcgga caagcttatt
gctcgtaaaa aagactggga
3661 tccaaaaaaa tatggtggtt ttgatagtcc aacggtagct
tattcagtcc tagtggttgc
3721 taaggtggaa aaagggaaat cgaagaagtt aaaatccgtt
aaagagttac tagggatcac
3781 aattatggaa agaagttcct ttgaaaaaaa tccgattgac
tttttagaag ctaaaggata
3841 taaggaagtt aaaaaagact taatcattaa actacctaaa
tatagtcttt ttgagttaga
3901 aaacggtcgt aaacggatgc tggctagtgc cggagaatta
caaaaaggaa atgagctggc
3961 tctgccaagc aaatatgtga attttttata tttagctagt
cattatgaaa agttgaaggg
4021 tagtccagaa gataacgaac aaaaacaatt gtttgtggag
cagcataagc attatttaga
4081 tgagattatt gagcaaatca gtgaattttc taagcgtgtt
attttagcag atgccaattt
4141 agataaagtt cttagtgcat ataacaaaca tagagacaaa
ccaatacgtg aacaagcaga
4201 aaatattatt catttattta cgttgacgaa tcttggagct
cccgctgctt ttaaatattt
4261 tgatacaaca attgatcgta aacgatatac gtctacaaaa
gaagttttag atgccactct
4321 tatccatcaa tccatcactg gtctttatga aacacgcatt
gatttgagtc agctaggagg
4381 tgactgaccc gggggaactg gccgtcgttt tacaacgtcg
tgactgggaa aaccctggcg
4441 ttacccaact taatcgcctt gcagcacatc cccctttcgc
cagctggcgt aatagcgaag
4501 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
gaatggcgaa tggcgcctga
4561 tgcggtattt tctccttacg catctgtgcg gtatttcaca
ccgcatatgg tgcactctca
4621 gtacaatctg ctctgatgcc gcatagttaa gccagccccg
acacccgcca acacccgctg
4681 acgcgccctg acgggcttgt ctgctcccgg catccgctta
cagacaagct gtgaccgtct
4741 ccgggagctg catgtgtcag aggttttcac cgtcatcacc
gaaacgcgcg agacgaaagg
4801 gcctcgtgat acgcctattt ttataggtta atgtcatgat
aataatggtt tcttagacgt
4861 caggtggcac ttttcgggga aatgtgcgcg gaacccctat
ttgtttattt ttctaaatac
4921 attcaaatat gtatccgctc atgagacaat aaccctgata
aatgcttcaa taatattgaa
4981 aaaggaagag tatgagtatt caacatttcc gtgtcgccct
tattcccttt tttgcggcat
5041 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
agtaaaagat gctgaagatc
5101 agttgggtgc acgagtgggt tacatcgaac tggatctcaa
cagcggtaag atccttgaga
5161 gttttcgccc cgaagaacgt tttccaatga tgagcacttt
taaagttctg ctatgtggcg
5221 cggtattatc ccgtattgac gccgggcaag agcaactcgg
tcgccgcata cactattctc
5281 agaatgactt ggttgagtac tcaccagtca cagaaaagca
tcttacggat ggcatgacag
5341 taagagaatt atgcagtgct gccataacca tgagtgataa
cactgcggcc aacttacttc
5401 tgacaacgat cggaggaccg aaggagctaa ccgctttttt
gcacaacatg ggggatcatg
5461 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
cataccaaac gacgagcgtg
5521 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
actattaact ggcgaactac
5581 ttactctagc ttcccggcaa caattaatag actggatgga
ggcggataaa gttgcaggac
5641 cacttctgcg ctcggccctt ccggctggct ggtttattgc
tgataaatct ggagccggtg
5701 agcgtgggtc tcgcggtatc attgcagcac tggggccaga
tggtaagccc tcccgtatcg
5761 tagttatcta cacgacgggg agtcaggcaa ctatggatga
acgaaataga cagatcgctg
5821 agataggtgc ctcactgatt aagcattggt aactgtcaga
ccaagtttac tcatatatac
5881 tttagattga tttaaaactt catttttaat ttaaaaggat
ctaggtgaag atcctttttg
5941 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
ccactgagcg tcagaccccg
6001 tagaaaagat caaaggatct tcttgagatc ctttttttct
gcgcgtaatc tgctgcttgc
6061 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
ggatcaagag ctaccaactc
6121 tttttccgaa ggtaactggc ttcagcagag cgcagatacc
aaatactgtc cttctagtgt
6181 agccgtagtt aggccaccac ttcaagaact ctgtagcacc
gcctacatac ctcgctctgc
6241 taatcctgtt accagtggct gctgccagtg gcgataagtc
gtgtcttacc gggttggact
6301 caagacgata gttaccggat aaggcgcagc ggtcgggctg
aacggggggt tcgtgcacac
6361 agcccagctt ggagcgaacg acctacaccg aactgagata
cctacagcgt gagctatgag
6421 aaagcgccac gcttcccgaa gggagaaagg cggacaggta
tccggtaagc ggcagggtcg
6481 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
ctggtatctt tatagtcctg
6541 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
atgctcgtca ggggggcgga
6601 gcctatggaa aaacgccagc aacgcggcct ttttacggtt
cctggccttt tgctggcctt
6661 ttgctcacat gttctttcct gcgttatccc ctgattctgt
ggataaccgt attaccgcct
6721 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
gcgcagcgag tcagtgagcg
6781 aggaagcgga agagcgccca atacgcaaac cgcctctccc
cgcgcgttgg ccgattcatt
6841 aatgcagctg gcacgacagg tttcccgact ggaaagcggg
cagtgagcgc aacgcaatta
6901 atgtgagtta gctcactcat taggcacccc aggctttaca
ctttatgctt ccggctcgta
6961 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
aaacagctat gaccatgatt
7021 acgccaagct ttggctaaca cacacgccat tccaaccaat
agttttctcg gcataaagcc
7081 atgctctgac gcttaaatgc actaatgcct taaaaaaaca
ttaaagtcta acacactaga
7141 cttatttact tcgtaattaa gtcgttaaac cgtgtgctct
acgaccaaaa gtataaaacc
7201 tttaagaact ttcttttttc ttgtaaaaaa agaaactaga
taaatctctc atatctttta
7261 ttcaataatc gcatcagatt gcagtataaa tttaacgatc
actcatcatg ttcatattta
7321 tcagagctcg tgctataatt atactaattt tataaggagg
aaaaaataaa gagggttata
7381 atgaacgaga aaaatataaa acacagtcaa aactttatta
cttcaaaaca taatatagat
7441 aaaataatga caaatataag attaaatgaa catgataata
tctttgaaat cggctcagga
7501 aaagggcatt ttacccttga attagtacag aggtgtaatt
tcgtaactgc cattgaaata
7561 gaccataaat tatgcaaaac tacagaaaat aaacttgttg
atcacgataa tttccaagtt
7621 ttaaacaagg atatattgca gtttaaattt cctaaaaacc
aatcctataa aatatttggt
7681 aatatacctt ataacataag tacggatata atacgcaaaa
ttgtttttga tagtatagct
7741 gatgagattt atttaatcgt ggaatacggg tttgctaaaa
gattattaaa tacaaaacgc
7801 tcattggcat tatttttaat ggcagaagtt gatatttcta
tattaagtat ggttccaaga
7861 gaatattttc atcctaaacc taaagtgaat agctcactta
tcagattaaa tagaaaaaaa
7921 tcaagaatat cacacaaaga taaacagaag tataattatt
tcgttatgaa atgggttaac
7981 aaagaataca agaaaatatt tacaaaaaat caatttaaca
attccttaaa acatgcagga
8041 attgacgatt taaacaatat tagctttgaa caattcttat
ctcttttcaa tagctataaa
8101 ttatttaata agtaagttaa gggatgcata aactgcatcc
cttaacttgt ttttcgtgta
8161 cctatttttt gtgaatcgat tatgtctttt gcgcattcac
ttcttttcta tataaatatg
8221 agcgaagcga ataagcgtcg gaaaagcagc aaaaagtttc
ctttttgctg ttggagcatg
8281 ggggttcagg gggtgcagta tctgacgtca atgccgagcg
aaagcgagcc gaagggtagc
8341 atttacgtta gataaccccc tgatatgctc cgacgcttta
tatagaaaag aagattcaac
8401 taggtaaaat cttaatatag gttgagatga taaggtttat
aaggaatttg tttgttctaa
8461 tttttcactc attttgttct aatttctttt aacaaatgtt
cttttttttt tagaacagtt
8521 atgatatagt tagaatagtt taaaataagg agtgagaaaa
agatgaaaga aagatatgga
8581 acagtctata aaggctctca gaggctcata gacgaagaaa
gtggagaagt catagaggta
8641 gacaagttat accgtaaaca aacgtctggt aacttcgtaa
aggcatatat agtgcaatta
8701 ataagtatgt tagatatgat tggcggaaaa aaacttaaaa
tcgttaacta tatcctagat
8761 aatgtccact taagtaacaa tacaatgata gctacaacaa
gagaaatagc aaaagctaca
8821 ggaacaagtc tacaaacagt aataacaaca cttaaaatct
tagaagaagg aaatattata
8881 aaaagaaaaa ctggagtatt aatgttaaac cctgaactac
taatgagagg cgacgaccaa
8941 aaacaaaaat acctcttact cgaatttggg aactttgagc
aagaggcaaa tgaaatagat
9001 tgacctccca ataacaccac gtagttattg ggaggtcaat
ctatgaaatg cgatta
//
