| 质粒名称: | pSL1142 (pSPIN, pBBR1 backbone) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 160730 |
| 存储实验室: | Samuel H. Sternberg |
|
载体骨架基本信息 |
|
| 载体名称: | pBBR1 |
| 载体抗性: | 卡那霉素 |
| 载体类型: | 细菌表达 |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | VchINTEGRATE proteins, crRNA, and donor DNA |
| 物种来源: | V. cholerae |
| 启动子 | J23119 |
| 克隆方法: | Gibson克隆 |
| 5' 测序引物 | cgccttcttgacgagttc |
| 3' 测序引物 | GCTAGTTATTGCTCAGCGG |
|
文献引用方法 |
|
| 材料和方法部分: | pSL1142 (pSPIN, pBBR1 backbone) was a gift from Samuel H. Sternberg (Addgene plasmid # 160730 ; http://n2t.net/addgene:160730 ; RRID:Addgene_160730) |
| 参考文献部分: | CRISPR RNA-guided integrases for high-efficiency, multiplexed bacterial genome engineering. Vo PLH, Ronda C, Klompe SE, Chen EE, Acree C, Wang HH, Sternberg SH. Nat Biotechnol. 2020 Nov 23. pii: 10.1038/s41587-020-00745-y. doi: 10.1038/s41587-020-00745-y. 10.1038/s41587-020-00745-y PubMed 33230293 |
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
质粒图谱.png)
载体简介
载体序列
LOCUS pSL1142 (pSPIN, pBBR1 backbone) 14031 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Single-plasmid V. cholerae INTEGRATE system, encodes all proteins,
crRNA, and donor DNA. Entry vector encodes non-targeting crRNA, with
BsaI sites for spacer cloning. pBBR1 backbone..
ACCESSION .
VERSION .
KEYWORDS pSL1142 (pSPIN, pBBR1 backbone)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14031)
AUTHORS Vo PLH, Ronda C, Klompe SE, Chen EE, Acree C, Wang HH, Sternberg SH
TITLE CRISPR RNA-guided integrases for high-efficiency, multiplexed
bacterial genome engineering.
JOURNAL Nat Biotechnol. 2020 Nov 23. pii: 10.1038/s41587-020-00745-y. doi:
10.1038/s41587-020-00745-y.
PUBMED 33230293
REFERENCE 2 (bases 1 to 14031)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..14031
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 122..893
/label=pBBR1 oriV
/note="replication origin of the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica; requires the pBBR1
Rep protein for replication"
CDS 2373..3167
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2427..2446)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3037..3056
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 3611..3628
/label=L4440
/note="L4440 vector, forward primer"
promoter 3639..3673
/label=J23119 promoter
/note="bacterial promoter (Registry of Standard Biological
Parts BBa_J23119)"
CDS 10655..10666
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(12101..12119)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 12115..12162
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS 12317..12976
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(12383..12402)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
primer_bind complement(13176..13193)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(13176..13192)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(13185..13207)
/label=M13/pUC Forward
/note="In lacZ gene"
CDS complement(13490..121)
/codon_start=1
/product="replication protein for the broad-host-range
plasmid pBBR1 from Bordetella bronchiseptica"
/label=pBBR1 Rep
/translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
ORIGIN
1 ccgccgtggg tttcctcgca ataagcccgg cccacgcctc atgcgctttg cgttccgttt
61 gcacccagtg accgggcttg ttcttggctt gaatgccgat ttctctggac tgcgtggcca
121 tgcttatctc catgcggtag gggtgccgca cggttgcggc accatgcgca atcagctgca
181 acttttcggc agcgcgacaa caattatgcg ttgcgtaaaa gtggcagtca attacagatt
241 ttctttaacc tacgcaatga gctattgcgg ggggtgccgc aatgagctgt tgcgtacccc
301 ccttttttaa gttgttgatt tttaagtctt tcgcatttcg ccctatatct agttctttgg
361 tgcccaaaga agggcacccc tgcggggttc ccccacgcct tcggcgcggc tccccctccg
421 gcaaaaagtg gcccctccgg ggcttgttga tcgactgcgc ggccttcggc cttgcccaag
481 gtggcgctgc ccccttggaa cccccgcact cgccgccgtg aggctcgggg ggcaggcggg
541 cgggcttcgc ccttcgactg cccccactcg cataggcttg ggtcgttcca ggcgcgtcaa
601 ggccaagccg ctgcgcggtc gctgcgcgag ccttgacccg ccttccactt ggtgtccaac
661 cggcaagcga agcgcgcagg ccgcaggccg gaggcttttc cccagagaaa attaaaaaaa
721 ttgatggggc aaggccgcag gccgcgcagt tggagccggt gggtatgtgg tcgaaggctg
781 ggtagccggt gggcaatccc tgtggtcaag ctcgtgggca ggcgcagcct gtccatcagc
841 ttgtccagca gggttgtcca cgggccgagc gaagcgagcc agccggtggc cgctcgcggc
901 catcgtccac atatccacgg gctggcaagg gagcgcagcg accgcgcagg gcgaagcccg
961 gagagcaagc ccgtagggcg ccgcagccgc cgtaggcggt cacgactttg cgaagcaaag
1021 tctagtgagt atactcaagc attgagtggc ccgccggagg caccgccttg cgctgccccc
1081 gtcgagccgg ttggacacca aaagggaggg gcaggcatgg cggcatacgc gatcatgcga
1141 tgcaagaagc tggcgaaaat gggcaacgtg gcggccagtc tcaagcacgc ctaccgcgag
1201 cgcgagacgc ccaacgctga cgccagcagg acgccagaga acgagcactg ggcggccagc
1261 agcaccgatg aagcgatggg ccgactgcgc gagttgctgc cagagaagcg gcgcaaggac
1321 gctgtgttgg cggtcgagta cgtcatgacg gccagcccgg aatggtggaa gtcggccagc
1381 caagaacagc aggcggcgtt cttcgagaag gcgcacaagt ggctggcgga caagtacggg
1441 gcggatcgca tcgtgacggc cagcatccac cgtgacgaaa ccagcccgca catgaccgcg
1501 ttcgtggtgc cgctgacgca ggacggcagg ctgtcggcca aggagttcat cggcaacaaa
1561 gcgcagatga cccgcgacca gaccacgttt gcggccgctg tggccgatct agggctgcaa
1621 cggggcatcg agggcagcaa ggcacgtcac acgcgcattc aggcgttcta cgaggccctg
1681 gagcggccac cagtgggcca cgtcaccatc agcccgcaag cggtcgagcc acgcgcctat
1741 gcaccgcagg gattggccga aaagctggga atctcaaagc gcgttgagac gccggaagcc
1801 gtggccgacc ggctgacaaa agcggttcgg caggggtatg agcctgccct acaggccgcc
1861 gcaggagcgc gtgagatgcg caagaaggcc gatcaagccc aagagacggc ccgtgacctt
1921 cgggagcgcc tgaagcccgt tctggacgcc ctggggccgt tgaatcggga tatgcaggcc
1981 aaggccgccg cgatcatcaa ggccgtgggc gaaaagctgc tgacggaaca gcgggaagtc
2041 cagcgccaga aacaggccca gcgccagcag gaacgcgggc gcgcacattt ccccgaaaag
2101 tgccacctgg gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa
2161 agagaaagca ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat
2221 ggacagcaag cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct
2281 gcaaagtaaa ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagat
2341 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
2401 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
2461 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
2521 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
2581 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
2641 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
2701 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
2761 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
2821 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
2881 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
2941 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
3001 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
3061 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
3121 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
3181 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
3241 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
3301 ccagcgcggg gatctcatgc tggagttctt cgcccacccc catgggcaaa tattatacgc
3361 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
3421 tccatgtcgg cagaatgctt aatgaattac aacagttttt atgcatgcgc ccaatacgca
3481 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
3541 actggtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
3601 gaccgagcgc agcgagtcag tgagcgagga cacaagcttt gacagctagc tcagtcctag
3661 gtataatact agcgtcgacg tggagatata ccatgggtga actgccgagt aggtagctga
3721 taacgagacc tctgtcttgt cagctagggt ggtctcgtga actgccgagt aggtagctga
3781 taacggatcc gaattcgagc gaaggagata tacatatgtt tttgcaaaga cctaaacctt
3841 acagcgatga aagtttagaa agtttcttta tccgagtggc taacaaaaat ggctacggtg
3901 atgtccatcg cttcctagaa gccactaaac gattccttca agacattgac cataatggct
3961 atcaaacctt tccgactgat ataactcgga taaacccata ctcagctaaa aacagttcca
4021 gcgcacgaac tgcgtcattc ctgaaacttg cacaattgac atttaatgaa ccgccagagc
4081 tacttgggtt ggcaattaac agaacaaaca tgaaatactc gccgtcaact agcgcggttg
4141 ttagaggtgc agaagtcttt cctcgcagtt tactacggac gcactccatc ccctgctgtc
4201 ctttgtgtct gcgagaaaat ggctacgcct cctacctttg gcactttcag gggtacgaat
4261 actgccacag ccataacgta cctttaatta ccacttgtag ctgtggtaag gagtttgact
4321 accgagtatc tgggttaaag ggcatttgct gcaaatgcaa ggagcctatc accttaacca
4381 gcagggagaa cggtcatgag gcagcgtgta ctgtttcaaa ctggcttgct ggccatgaat
4441 ctaaacctct gccaaatctt cctaaaagct accgatgggg tttagttcat tggtggatgg
4501 gtattaaaga tagcgagttc gatcactttt cgttcgttca atttttctca aactggccaa
4561 ggtcattcca ctcgataatc gaagatgaag tagagttcaa ccttgagcat gctgttgtca
4621 gcacgtctga attacgacta aaagatcttc ttggtcgatt gtttttcggt tcaattcggt
4681 tacctgagcg gaatcttcaa cacaatatca tccttggtga gcttctctgc tatttagaaa
4741 atcgtttatg gcaagacaag ggattaatcg ccaacctcaa aatgaacgcg ttagaggcga
4801 ctgtaatgtt aaattgtagc ctcgatcaga ttgcatcaat ggttgaacaa cgcatcttga
4861 agccaaatcg aaaaagcaag cccaacagcc ctcttgatgt taccgattat ctatttcatt
4921 tcggcgatat tttctgtctt tggttagctg agttccaaag cgatgagttt aaccgttcgt
4981 tttatgtgtc gaggtggtaa atgcaaactc tgaaagaact aatcgcatcc aatcctgacg
5041 acttaacaac tgaacttaag agagcatttc gtccactcac accgcatatt gcaattgatg
5101 gtaatgaact tgacgcactg acgatattag tcaatttaac cgataagact gatgatcaga
5161 aagacctgct cgatcgagcc aaatgcaagc aaaaacttcg agatgaaaaa tggtgggcta
5221 gctgcataaa ttgcgttaat tacagacaaa gccataaccc aaaattcccg gatatacgtt
5281 ctgaaggcgt gattcgaacc caagccctgg gtgaattacc gagtttccta ctctcgtcct
5341 caaaaatccc accataccat tggtcatata gccatgattc aaaatacgtc aacaaaagcg
5401 cattcctcac caatgagttt tgttgggatg gtgagatctc atgtttgggt gagcttctta
5461 aagatgcaga tcacccactt tggaatactt tgaaaaagtt aggttgttct caaaaaactt
5521 gcaaagcaat ggcaaaacaa ctagctgata ttactctcac gactatcaat gtcacactcg
5581 caccaaatta cctgactcaa atatctctcc ccgatagtga tacatcctac atttcactct
5641 ctcctgttgc atcgctatcg atgcaaagcc actttcatca gaggctccaa gatgaaaatc
5701 ggcatagcgc gataacgcgg tttagccgaa ctaccaacat gggggtcact gcgatgacat
5761 gtggcggcgc atttagaatg ttaaagtctg gcgctaagtt ttccagcccc cctcatcacc
5821 gattaaacag taaacgaagt tggttgacgt cagagcatgt tcagtcatta aaacagtacc
5881 agcgcctcaa caaaagcctc atacctgaaa actctcggat tgcactccgt agaaaataca
5941 aaatcgagct tcaaaatatg gtcagatctt ggtttgcaat gcaagaccat acccttgatt
6001 cgaacatact tatccaacat ttgaatcatg acctatctta cttaggagcc acaaaacgtt
6061 ttgcatacga tccagcgatg accaagctct ttactgagct tttgaaacga gagttatcaa
6121 attcaatcaa taatggtgag caacacacta atggatcgtt tttagtccta ccgaatatca
6181 gggtttgtgg cgcaacagct ttaagctccc cggtaacggt ggggattcca tcacttacag
6241 ctttctttgg cttcgttcac gcatttgaac ggaatataaa tcgcaccacc tcatcgtttc
6301 gtgttgaatc ctttgcgata tgcgtccatc aactacatgt cgagaagcga ggtttgacag
6361 cagagtttgt ggaaaaaggc gacgggacta tatccgctcc cgcgacccgg gatgactggc
6421 agtgtgatgt cgtatttagc cttattttga acaccaactt tgctcaacat attgaccaag
6481 atacgttagt tacatcacta ccaaagcgat tggctcgggg ttcagcaaaa attgcgattg
6541 atgactttaa acatatcaac tcattctcga cattagaaac agcgatcgaa tctctgccaa
6601 tagaagctgg taggtggtta tcactttacg cacagtcaaa caataatcta agtgatctat
6661 tagcagccat gacagaggac catcagctca tggcaagctg cgtcggttac cacttgttag
6721 aagagcccaa agataaacca aactccctca gaggttacaa acacgctatc gccgagtgca
6781 tcattggact cattaactca atcaccttta gctcagagac tgatcccaac acaatctttt
6841 ggtcgctaaa gaactatcaa aactacctag tggtacagcc aaggagtatc aacgatgaaa
6901 ctaccgacaa atctagccta tgagcgctct atcgacccat cagatgtctg tttttttgtc
6961 gtctggcccg atgatagaaa aacaccttta acctacaatt ctcgtactct gctcgggcaa
7021 atggaagcgg catcattagc ctatgatgtc tcaggtcaac caataaaaag tgccaccgct
7081 gaggcgttag ctcaagggaa ccctcatcaa gttgatttct gccacgttcc atacggtgcg
7141 agtcatattg aatgcagttt ctccgtctcg ttttcttctg aactacgtca accatataag
7201 tgtaactcaa gcaaagttaa acaaacgcta gtgcaattag tcgagctcta cgaaacgaaa
7261 atcggctgga ctgagctagc aacccgatat ttgatgaaca tttgcaacgg taaatggctg
7321 tggaaaaata cccgtaaagc ctattgctgg aacattgtac ttacaccttg gccttggaac
7381 ggggaaaagg ttggatttga agatattcgt actaactaca cctcacggca agactttaaa
7441 aataataaaa attggtctgc tatagttgaa atgatcaaaa ccgcattttc tagtactgat
7501 gggctggcga tatttgaagt cagggccacc ttgcacttgc caacgaatgc tatggtgcgg
7561 ccaagccaag ttttcacaga aaaagaaagt ggcagtaaaa gtaaatctaa aactcaaaac
7621 agtcgagttt ttcagagtac aactattgat ggtgaacgat cgccaatact aggggccttt
7681 aaaacgggag cagctattgc aaccattgac gactggtatc ctgaagccac tgagccacta
7741 agggtcggac ggtttggggt tcatcgcgaa gatgtcactt gctaccgtca tccgtctacc
7801 ggaaaagatt ttttctcgat attacaacaa gcagagcact atattgaagt gttgagcgcc
7861 aacaaaactc ccgctcaaga aactatcaac gacatgcact ttttaatggc taacctgatt
7921 aagggtggga tgttccagca taaaggagac tgactgtgaa atggtattat aagacaatca
7981 cctttctgcc agagttgtgc aacaacgagt cactggctgc aaagtgtctc cgcgttctgc
8041 atggatttaa ctatcagtat gagacacgaa atatcggcgt ttcatttccg ctttggtgtg
8101 atgcaacggt tggaaaaaag atttcatttg tcagcaagaa caagatagaa ctcgacttac
8161 tacttaaaca acactatttc gtccaaatgg aacaacttca atactttcat atatccaaca
8221 ctgttctcgt cccagaagat tgtacatacg tttcctttag acgctgtcaa tctatagata
8281 agctcacagc agcagggctg gcaaggaaaa tcagacgcct ggagaaacgt gctctatcta
8341 gaggcgagca atttgaccca tcatcttttg ctcaaaaaga gcatactgca atagcgcact
8401 accactcact tggggagtcc agcaaacaga cgaaccgcaa ctttcgactc aatatcagga
8461 tgctctcgga gcaaccccgt gaggggaact cgatttttag tagctatggc ttatcaaatt
8521 cagaaaactc gtttcagcct gtacccttaa tctgaccttt aataaggaga tataccatgg
8581 cgacaagttt acctacgccc tcagcaatta cgacttcggc gttagagtat gcattccata
8641 ctcccgctcg caatctaacg aaatctcgcg gaaaaaacat tcatcgttat gtcagtgtaa
8701 agatgagtaa gaggattacg gtagaatcta ctctagagtg tgatgcctgc tatcactttg
8761 attttgagcc aagtattgtt cgcttttgcg ctcaaccgat tcgattttta tattatctca
8821 atggtcagtc tcactcctat gttcctgact ttctagttca atttgatacc aacgagtttg
8881 ttctatatga agtaaagtca gcttatgcta agaacaaacc tgattttgat gttgaatggg
8941 aggcgaaagt aaaagcagca actgaactag ggctagaatt ggagcttgtt gaagagagtg
9001 atattaggga tacggttgta ttaaataatc ttaagcgcat gcatcgttat gcttcgaaag
9061 atgagctgaa taacgtacat aactctctct taaaaataat aaagtacaat ggcgcccaat
9121 ctgcaagatg cttgggagaa cagttgggtt taaaaggccg aactgtttta ccaattttgt
9181 gcgatttgct gtcaaggtgt ttactcgata cacgtttgga taagcctcta tctcttgaat
9241 ctcgatttga gttggccagt tatggctaag aaagggttct caagtttcca tagaaaagca
9301 gtctcttccc aagatacgct tgaatcgata gagcttgtct ctagcgctaa ttgtctagaa
9361 agtgttacgt atcaagatat atcagcattt cccgaaacaa ttgcggtaga gattaatttc
9421 cgattaagca ttcttcgctt tttagcgcga aagtgcgaaa ccattgtggc caaatcaatt
9481 gaaccacatc gtgtagagct acagcaaaac tatagtagaa aaatacccag tgcaataacg
9541 atatatcgat ggtggcttgc ttttcgaaaa tcagactaca accccattag cttagcacct
9601 aatatcaagg atagaggtaa tagagagaca aaagtgtcaa cagttgttga ttctattatg
9661 gaacaggcag ttgaaagagt tatatctgga cgaaaagtca atgttagctc tgcatataaa
9721 cgtgttcgac gaaaagttcg tcaatacaat ctaactcatg gaacgaaata cacgtatcct
9781 aagtacgaat ctgtaagaaa gcgagtaaaa aagaaaaccc catttgagtt attagccgca
9841 gggaaagggg agagagtagc taagagagag tttcgccgaa tgggaaaaaa gatcctcacg
9901 tctagcgtgc tagagagggt tgaaatagat cacactgtcg ttgacctttt tgcagtacac
9961 gaagagtatc gaatcccatt gggccgacct tggcttactc aattggttga ttgttacagt
10021 aaagctgtaa tcggttttta tttaggtttc gagcctccta gctatgtgtc ggtttccctt
10081 gcacttaaga atgcaataca acgcaaagat gacttaatct cctcgtatga atcgatcgag
10141 aatgaatggc tatgttatgg catcccagac ctactcgtaa ctgataatgg taaagagttt
10201 ttgtcgaaag catttgatca agcatgtgaa tcactattga tcaatgtgca tcaaaataaa
10261 gttgagacgc ccgacaacaa acctcatgtt gaacgtaact acgggactat taatacttct
10321 ctgttagacg atttacctgg gaaatccttc agccagtacc ttcaaagaga agggtacgac
10381 tctgtgggag aagctaccct tacactcaat gagattagag aaatttactt aatttggttg
10441 gtggatattt atcataaaaa acccaatcag agaggcacta attgtcctaa tgttgcgtgg
10501 aaaaagggtt gtcaagaatg ggaaccagag gagttctctg gttctaaaga cgaattagac
10561 tttaaatttg ctattgttga ttacaaacaa cttactaaag tagggataac tgtctacaaa
10621 gaactgagtt atagcaatga ccgtttagct gaatatagag ggaagaaagg aaaccataaa
10681 gttcagttca agtataaccc tgagtgtatg gcagttattt gggtgttgga tgaggatatg
10741 aatgagtact ttacagttaa tgcgattgac tacgaatatg caagtagagt atcactttgg
10801 caacataaat ataacatgaa atatcaagca gaactaaatt cagcagaata tgatgaggac
10861 aaggaaattg atgcagaaat aaaaattgaa gaaatcgcag atcgttcaat tgttaagact
10921 aacaaaatca gagctcggag gcgtggcgct aggcatcaag agaatagcgc aagggctaag
10981 tcaatcagta atgcgaaccc ggcctcgata caaaaacatg aagatgaaat cgttagtgca
11041 gataatgacg attgggatat tgattatgtc tgagaaatcg tcaaatgagt gaaacgcgtg
11101 aggctcgtat atcaagagct aaaagggcat ttgtatccac accgagcgtt aggaaaatct
11161 tgagttacat ggatagatgt agagatctat cagacctaga gtctgagcct acatgcatga
11221 tggtctatgg tgcttcgggt gtaggtaaaa cgaccgtcat caagaaatac ttaaatcaaa
11281 acagaagaga gtccgaagcc gggggcgata taataccggt tttgcatatt gagttgccag
11341 acaatgcgaa gccagtagat gcagcaaggg aattgctggt tgaaatgggt gacccgctag
11401 cactttatga aactgactta gctagattga cgaaaagact gactgaatta atccctgcgg
11461 tcggcgtgaa gctgattatt atcgatgagt tccaacattt ggtggaagaa aggtcaaatc
11521 gggttcttac ccaagtaggt aattggctaa aaatgatact taacaaaacg aaatgtccaa
11581 ttgttatatt tggtatgcca tactcaaaag ttgtactgca agcaaactcg caacttcacg
11641 ggcgattttc cattcaggtt gaactgcgcc cctttagcta ccagggaggt agaggtgtat
11701 ttaaaacttt tttggaatac cttgataaag ccctaccttt tgaaaaacag gctggcttag
11761 ccaacgaaag tttgcagaaa aaattgtatg cattctctca gggaaacatg cgttcgttga
11821 gaaaccttat ttatcaagca tctatcgaag caattgataa tcagcatgag acgataaccg
11881 aagaagattt cgtttttgca tcgaagttga catcgggcga taaacccaac tcatggaaaa
11941 atccttttga ggagggtgtt gaggtaacag aagatatgtt acgaccgcca ccaaaagata
12001 ttggttggga ggactatttg agacattcaa ccccgagagt gagtaaacca ggtagaaata
12061 aaaacttttt cgaataacct aggctgctgc gctgctgcca ccgctgagca ataactagca
12121 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaacc tcaggcattt
12181 gttgttgata caaccataaa atgataatta cacccataaa ttgataatta tcacacccat
12241 aaattgatat tgcctcttca tggtctaaac ttcagtaagt ttacgacatt ttcctcgagg
12301 tcatttccgg ggatccatgg agaaaaaaat cactggatat accaccgttg atatatccca
12361 atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca
12421 gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt
12481 ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg agttccgtat
12541 ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt
12601 ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca
12661 gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc
12721 taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag
12781 ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca ctatgggcaa
12841 atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt
12901 ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg
12961 gcagggcggg gcgtaatttt tttaaggcag ttattggtgc ccttctagag tccttactgc
13021 agtagttttg ctgaaatact cgattcacaa aaatatcaac ttatggttgt tttgtgagat
13081 atcaatatat ggttgttttg tggttaagtt gctgattata aataattatt aaatatcact
13141 ttatggttgc atcaacaggt accgagctcg aattcactgg ccgtcgtttt acaacgtcgt
13201 gactgggaaa accctggcgt tacccaactt aatcgccttg cggccagttt gctcaggctc
13261 tccccgtgga ggtaataatt gacgatatga tcatttattc tgcctcccag agcctgataa
13321 aaacggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct
13381 ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcga ggcggctaca
13441 gccgatagtc tggaacagcg cacttacggg ttgctgcgca acccaagtgc taccggcgcg
13501 gcagcgtgac ccgtgtcggc ggctccaacg gctcgccatc gtccagaaaa cacggctcat
13561 cgggcatcgg caggcgctgc tgcccgcgcc gttcccattc ctccgtttcg gtcaaggctg
13621 gcaggtctgg ttccatgccc ggaatgccgg gctggctggg cggctcctcg ccggggccgg
13681 tcggtagttg ctgctcgccc ggatacaggg tcgggatgcg gcgcaggtcg ccatgcccca
13741 acagcgattc gtcctggtcg tcgtgatcaa ccaccacggc ggcactgaac accgacaggc
13801 gcaactggtc gcggggctgg ccccacgcca cgcggtcatt gaccacgtag gccgacacgg
13861 tgccggggcc gttgagcttc acgacggaga tccagcgctc ggccaccaag tccttgactg
13921 cgtattggac cgtccgcaaa gaacgtccga tgagcttgga aagtgtcttc tggctgacca
13981 ccacggcgtt ctggtggccc atctgcgcca cgaggtgatg cagcagcatt g
//
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