| 质粒名称: | pREDKI |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 51626 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pKD46 |
| 载体出品公司: | -- |
| 载体抗性: | 低卡那霉素 |
| 空载体大小: | 6300 bp |
| 完整质粒大小: | -- |
| 载体修饰: | replace amp to kan |
| 载体类型: | 细菌表达；红色重组酶和I-SceI核酸内切酶 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 30 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | I-SceI,mitochondrial intron of Saccharomyces cerevisiae |
| 物种来源: | S. cerevisiae (budding yeast) |
| 插入大小: | 3000 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | pRha |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pREDKI was a gift from Sheng Yang (Addgene plasmid # 51626 ; http://n2t.net/addgene:51626 ; RRID:Addgene_51626) |
| 参考文献部分: | High-Efficiency Scarless Genetic Modification in Escherichia coli by Using Lambda Red Recombination and I-SceI Cleavage. Yang J, Sun B, Huang H, Jiang Y, Diao L, Chen B, Xu C, Wang X, Liu J, Jiang W, Yang S. Appl Environ Microbiol. 2014 Jul 1;80(13):3826-3834. Epub 2014 Apr 18. 10.1128/AEM.00313-14 PubMed 24747889 |
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 pREDKI 9028 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION E coli Genetic Modification.
ACCESSION .
VERSION .
KEYWORDS pREDKI
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9028)
AUTHORS Yang J, Sun B, Huang H, Jiang Y, Diao L, Chen B, Xu C, Wang X, Liu
J, Jiang W, Yang S
TITLE High-Efficiency Scarless Genetic Modification in Escherichia coli by
Using Lambda Red Recombination and I-SceI Cleavage.
JOURNAL Appl Environ Microbiol. 2014 Jul 1;80(13):3826-3834. Epub 2014 Apr
18.
PUBMED 24747889
REFERENCE 2 (bases 1 to 9028)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9028
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(19..897)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
promoter 924..1208
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
primer_bind 1131..1150
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
RBS 1235..1243
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
CDS 1252..1668
/codon_start=1
/gene="gam"
/product="inhibitor of the host RecBCD nuclease in the
lambda Red system"
/label=Gam
/note="protects the double-stranded DNA substrate for
recombination"
/translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
CDS 1674..2459
/codon_start=1
/gene="bet"
/product="single-stranded DNA binding recombinase in the
lambda Red system"
/label=Beta
/note="mediates pairing of a single-stranded DNA with a
complementary target"
/translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
CDS 2456..3136
/codon_start=1
/gene="exo"
/product="5' to 3' double-stranded DNA exonuclease in the
lambda Red system"
/label=Exo
/note="creates 3' single-stranded DNA tails"
/translation="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPR
SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
EQWR"
terminator 3137..3381
/label=-lambda tL3 terminator
/note="transcription terminator tL3 from phage lambda"
CDS complement(3414..4262)
/codon_start=1
/gene="E. coli rhaR"
/product="transcriptional activator of rhaR and rhaS"
/label=rhaR
/note="needed for rhamnose-dependent activation of the rhaB
promoter"
/translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
D"
CDS complement(4336..5172)
/codon_start=1
/gene="E. coli rhaS"
/product="positive regulator of the rhaB promoter
"
/label=rhaS
/note="activated by L-rhamnose"
/translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
promoter 5318..5436
/label=rhaB promoter
/note="promoter of the E. coli rhaBAD operon, conferring
tight induction with L-rhamnose and repression with
D-glucose in the presence of RhaR and RhaS (Giacalone et
al., 2006)"
CDS complement(6205..7155)
/codon_start=1
/gene="rep101"
/product="RepA protein needed for replication with the
pSC101 origin"
/label=Rep101
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
rep_origin 7203..7425
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
primer_bind complement(8180..8199)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 8191..9006
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(8264..8283)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
ORIGIN
1 aaatggttta aacatttatt atgacaactt gacggctaca tcattcactt tttcttcaca
61 accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaaata
121 gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct
181 caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc
241 taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac
301 gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg
361 tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc gccgcagtaa
421 caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc
481 gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa
541 gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg
601 acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg
661 aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg
721 atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag
781 cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac
841 cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact
901 tttcatactc ccgccattca gagaagaaac caattgtcca tattgcatca gacattgccg
961 tcactgcgtc ttttactggc tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag
1021 cattctgtaa caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa
1081 tcacggcaga aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc
1141 atttttatcc ataagattag cggatcctac ctgacgcttt ttatcgcaac tctctactgt
1201 ttctccatac ccgttttttt gggaattcga gctctaagga ggttataaaa aatggatatt
1261 aatactgaaa ctgagatcaa gcaaaagcat tcactaaccc cctttcctgt tttcctaatc
1321 agcccggcat ttcgcgggcg atattttcac agctatttca ggagttcagc catgaacgct
1381 tattacattc aggatcgtct tgaggctcag agctgggcgc gtcactacca gcagctcgcc
1441 cgtgaagaga aagaggcaga actggcagac gacatggaaa aaggcctgcc ccagcacctg
1501 tttgaatcgc tatgcatcga tcatttgcaa cgccacgggg ccagcaaaaa atccattacc
1561 cgtgcgtttg atgacgatgt tgagtttcag gagcgcatgg cagaacacat ccggtacatg
1621 gttgaaacca ttgctcacca ccaggttgat attgattcag aggtataaaa cgaatgagta
1681 ctgcactcgc aacgctggct gggaagctgg ctgaacgtgt cggcatggat tctgtcgacc
1741 cacaggaact gatcaccact cttcgccaga cggcatttaa aggtgatgcc agcgatgcgc
1801 agttcatcgc attactgatc gttgccaacc agtacggcct taatccgtgg acgaaagaaa
1861 tttacgcctt tcctgataag cagaatggca tcgttccggt ggtgggcgtt gatggctggt
1921 cccgcatcat caatgaaaac cagcagtttg atggcatgga ctttgagcag gacaatgaat
1981 cctgtacatg ccggatttac cgcaaggacc gtaatcatcc gatctgcgtt accgaatgga
2041 tggatgaatg ccgccgcgaa ccattcaaaa ctcgcgaagg cagagaaatc acggggccgt
2101 ggcagtcgca tcccaaacgg atgttacgtc ataaagccat gattcagtgt gcccgtctgg
2161 ccttcggatt tgctggtatc tatgacaagg atgaagccga gcgcattgtc gaaaatactg
2221 catacactgc agaacgtcag ccggaacgcg acatcactcc ggttaacgat gaaaccatgc
2281 aggagattaa cactctgctg atcgccctgg ataaaacatg ggatgacgac ttattgccgc
2341 tctgttccca gatatttcgc cgcgacattc gtgcatcgtc agaactgaca caggccgaag
2401 cagtaaaagc tcttggattc ctgaaacaga aagccgcaga gcagaaggtg gcagcatgac
2461 accggacatt atcctgcagc gtaccgggat cgatgtgaga gctgtcgaac agggggatga
2521 tgcgtggcac aaattacggc tcggcgtcat caccgcttca gaagttcaca acgtgatagc
2581 aaaaccccgc tccggaaaga agtggcctga catgaaaatg tcctacttcc acaccctgct
2641 tgctgaggtt tgcaccggtg tggctccgga agttaacgct aaagcactgg cctggggaaa
2701 acagtacgag aacgacgcca gaaccctgtt tgaattcact tccggcgtga atgttactga
2761 atccccgatc atctatcgcg acgaaagtat gcgtaccgcc tgctctcccg atggtttatg
2821 cagtgacggc aacggccttg aactgaaatg cccgtttacc tcccgggatt tcatgaagtt
2881 ccggctcggt ggtttcgagg ccataaagtc agcttacatg gcccaggtgc agtacagcat
2941 gtgggtgacg cgaaaaaatg cctggtactt tgccaactat gacccgcgta tgaagcgtga
3001 aggcctgcat tatgtcgtga ttgagcggga tgaaaagtac atggcgagtt ttgacgagat
3061 cgtgccggag ttcatcgaaa aaatggacga ggcactggct gaaattggtt ttgtatttgg
3121 ggagcaatgg cgatgacgca tcctcacgat aatatccggg taggcgcaat cactttcgtc
3181 tactccgtta caaagcgagg ctgggtattt cccggccttt ctgttatccg aaatccactg
3241 aaagcacagc ggctggctga ggagataaat aataaacgag gggctgtatg cacaaagcat
3301 cttctgttga gttaagaacg agtatcgaga tggcacatag ccttgctcaa attggaatca
3361 ggtttgtgcc aataccagta gaaacagacg aagaatccat ggggcatggc gaattaatct
3421 ttctgcgaat tgagatgacg ccactggctg ggcgtcatcc cggtttcccg ggtaaacacc
3481 accgaaaaat agttactatc ttcaaagcca cattcggtcg aaatatcact gattaacagg
3541 cggctatgct ggagaagata ttgcgcatga cacactctga cctgtcgcag atattgattg
3601 atggtcattc cagtctgctg gcgaaattgc tgacgcaaaa cgcgctcact gcacgatgcc
3661 tcatcacaaa atttatccag cgcaaaggga cttttcaggc tagccgccag ccgggtaatc
3721 agcttatcca gcaacgtttc gctggatgtt ggcggcaacg aatcactggt gtaacgatgg
3781 cgattcagca acatcaccaa ctgcccgaac agcaactcag ccatttcgtt agcaaacggc
3841 acatgctgac tactttcatg ctcaagctga ccgataacct gccgcgcctg cgccatcccc
3901 atgctaccta agcgccagtg tggttgccct gcgctggcgt taaatcccgg aatcgccccc
3961 tgccagtcaa gattcagctt cagacgctcc gggcaataaa taatattctg caaaaccaga
4021 tcgttaacgg aagcgtagga gtgtttatcg tcagcatgaa tgtaaaagag atcgccacgg
4081 gtaatgcgat aagggcgatc gttgagtaca tgcaggccat taccgcgcca gacaatcacc
4141 agctcacaaa aatcatgtgt atgttcagca aagacatctt gcggataacg gtcagccaca
4201 gcgactgcct gctggtcgct ggcaaaaaaa tcatctttga gaagttttaa ctgatgcgcc
4261 accgtggcta cctcggccag agaacgaagt tgattattcg caatatggcg tacaaatacg
4321 ttgagaagat tcgcgttatt gcagaaagcc atcccgtccc tggcgaatat cacgcggtga
4381 ccagttaaac tctcggcgaa aaagcgtcga aaagtggtta ctgtcgctga atccacagcg
4441 ataggcgatg tcagtaacgc tggcctcgct gtggcgtagc agatgtcggg ctttcatcag
4501 tcgcaggcgg ttcaggtatc gctgaggcgt cagtcccgtt tgctgcttaa gctgccgatg
4561 tagcgtacgc agtgaaagag aaaattgatc cgccacggca tcccaattca cctcatcggc
4621 aaaatggtcc tccagccagg ccagaagcaa gttgagacgt gatgcgctgt tttccaggtt
4681 ctcctgcaaa ctgcttttac gcagcaagag cagtaattgc ataaacaaga tctcgcgact
4741 ggcggtcgag ggtaaatcat tttccccttc ctgctgttcc atctgtgcaa ccagctgtcg
4801 cacctgctgc aatacgctgt ggttaacgcg ccagtgagac ggatactgcc catccagctc
4861 ttgtggcagc aactgattca gcccggcgag aaactgaaat cgatccggcg agcgatacag
4921 cacattggtc agacacagat tatcggtatg ttcatacaga tgccgatcat gatcgcgtac
4981 gaaacagacc gtgccaccgg tgatggtata gggctgccca ttaaacacat gaatacccgt
5041 gccatgttcg acaatcacaa tttcatgaaa atcatgatga tgttcaggaa aatccgcctg
5101 cgggagccgg ggttctatcg ccacggacgc gttaccagac ggaaaaaaat ccacactatg
5161 taatacggtc atactggcct cctgatgtcg tcaacacggc gaaatagtaa tcacgaggtc
5221 aggttcttac cttaaatttt cgacggaaaa ccacgtaaaa aacgtcgatt tttcaagata
5281 cagcgtgaat tttcaggaaa tgcggtgagc atcacatcac cacaattcag caaattgtga
5341 acatcatcac gttcatcttt ccctggttgc caatggccca ttttcctgtc agtaacgaga
5401 aggtcgcgaa ttcaggcgct ttttagactg gtcgtaatga aattcagcag gatcacatat
5461 gcatcaaaaa aaccaggtaa tgaacctggg tccgaactct aaactgctga aagaatacaa
5521 atcccagctg atcgaactga acatcgaaca gttcgaagca ggtatcggtc tgatcctggg
5581 tgatgcttac atccgttctc gtgatgaagg taaaacctac tgtatgcagt tcgagtggaa
5641 aaacaaagca tacatggacc acgtatgtct gctgtacgat cagtgggtac tgtctttcaa
5701 acaccaagct ttcaacaaac tggctaacct gttcatcgtt aacaacaaaa aaaccatccc
5761 gaacaacctg gttgaaaact acctgacccc gatgtctctg gcatactggt tcatggatga
5821 tggtggtaaa tgggattaca acaaaaactc taccaacaaa tcgatcgtac tgaacaccca
5881 gtctttcact ttcgaagaag tagaatacct ggttaagggt ctgcgtaaca aattccaact
5941 gaactgttac gtaaaaatca acaaaaacaa accgatcatc tacatcgatt ctatgtctta
6001 cctgatcttc tacaacctga tcaaaccgta cctgatcccg cagatgatgt acaaactgcc
6061 gaacactatc tcctccgaaa ctttcctgaa ataataacca tgggtatgga cagttttccc
6121 tttgatatgt aacggtgaac agttgttcta cttttgtttg ttagtcttga tgcttcactg
6181 atagatacaa gagccataag aacctcagat ccttccgtat ttagccagta tgttctctag
6241 tgtggttcgt tgtttttgcg tgagccatga gaacgaacca ttgagatcat acttactttg
6301 catgtcactc aaaaattttg cctcaaaact ggtgagctga atttttgcag ttaaagcatc
6361 gtgtagtgtt tttcttagtc cgttacgtag gtaggaatct gatgtaatgg ttgttggtat
6421 tttgtcacca ttcattttta tctggttgtt ctcaagttcg gttacgagat ccatttgtct
6481 atctagttca acttggaaaa tcaacgtatc agtcgggcgg cctcgcttat caaccaccaa
6541 tttcatattg ctgtaagtgt ttaaatcttt acttattggt ttcaaaaccc attggttaag
6601 ccttttaaac tcatggtagt tattttcaag cattaacatg aacttaaatt catcaaggct
6661 aatctctata tttgccttgt gagttttctt ttgtgttagt tcttttaata accactcata
6721 aatcctcata gagtatttgt tttcaaaaga cttaacatgt tccagattat attttatgaa
6781 tttttttaac tggaaaagat aaggcaatat ctcttcacta aaaactaatt ctaatttttc
6841 gcttgagaac ttggcatagt ttgtccactg gaaaatctca aagcctttaa ccaaaggatt
6901 cctgatttcc acagttctcg tcatcagctc tctggttgct ttagctaata caccataagc
6961 attttcccta ctgatgttca tcatctgagc gtattggtta taagtgaacg ataccgtccg
7021 ttctttcctt gtagggtttt caatcgtggg gttgagtagt gccacacagc ataaaattag
7081 cttggtttca tgctccgtta agtcatagcg actaatcgct agttcatttg ctttgaaaac
7141 aactaattca gacatacatc tcaattggtc taggtgattt taatcactat accaattgag
7201 atgggctagt caatgataat tactagtcct tttcctttga gttgtgggta tctgtaaatt
7261 ctgctagacc tttgctggaa aacttgtaaa ttctgctaga ccctctgtaa attccgctag
7321 acctttgtgt gttttttttg tttatattca agtggttata atttatagaa taaagaaaga
7381 ataaaaaaag ataaaaagaa tagatcccag ccctgtgtat aactcactac tttagtcagt
7441 tccgcagtat tacaaaagga tgtcgcaaac gctgtttgct cctctacaaa acagacctta
7501 aaaccctaaa ggcttaagta gcaccctcgc aagctcggtt gcggccgcaa tcgggcaaat
7561 cgctgaatat tccttttgtc tccgaccatc aggcacctga gtcgctgtct ttttcgtgac
7621 attcagttcg ctgcgctcac ggctctggca gtgaatgggg gtaaatggca ctacaggcgc
7681 cttttatgga ttcatgcaag gaaactaccc ataatacaag aaaagcccgt cacgggcttc
7741 tcagggcgtt ttatggcggg tctgctatgt ggtgctatct gactttttgc tgttcagcag
7801 ttcctgccct ctgattttcc agtctgacca cttcggatta tcccgtgaca ggtcattcag
7861 actggctaat gcacccagta aggcagcggt atcatcaacg gggtctgacg ctcagtggaa
7921 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
7981 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
8041 tgacagttac gtttaaacca tttcctgcag gggggggggg aaagccacgt tgtgtctcaa
8101 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
8161 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcttgc
8221 tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
8281 gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca
8341 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
8401 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
8461 cctgatgatg catggttact caccactgcg atccccggga aaacagcatt ccaggtatta
8521 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
8581 ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct
8641 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
8701 aatggctggc ctgttgaaca agtctggaaa gaaatgcata agcttttgcc attctcaccg
8761 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
8821 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
8881 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
8941 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
9001 ttctaatcag aattggttaa ttggttgg
//
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