| 质粒名称: | pMGS46 (ARF16-PB1-HA-P2A-OsTIR1) |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 126580 |
| 存储实验室: | Michael Guertin |
|
载体骨架基本信息 |
|
| 载体名称: | pMK232 |
| 载体出品公司: | Masato Kanemaki (Addgene plasmid # 72834) |
| 空载体大小: | 9590 bp |
| 完整质粒大小: | 10247bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | Puromycin |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | OsARF16-PB1 domain |
| 物种来源: | Oryza sativa |
| 插入大小: | 654 bp |
| 标签/融合蛋白: | HA (C terminal on backbone) |
| 克隆方法: | Ligation Independent Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | MGS46 (ARF16-PB1-HA-P2A-OsTIR1) was a gift from Michael Guertin (Addgene plasmid # 126580 ; http://n2t.net/addgene:126580 ; RRID:Addgene |
| 参考文献部分: | An improved auxin-inducible degron system preserves native protein levels and enables rapid and specific protein depletion. Sathyan KM, McKenna BD, Anderson WD, Duarte FM, Core L, Guertin MJ. Genes Dev. 2019 Oct 1;33(19-20):1441-1455. doi: 10.1101/gad.328237.119. Epub 2019 Aug 29. 10.1101/gad.328237.119 PubMed 31467088 |
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 pMGS46 (ARF16-PB1-HA-P2A-OsTIR1) 10247 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION A repair construct to express ARF16-PB1-HA and OsTIR1 under the
control of the CMV promoter from the human AAVS1 locus.
ACCESSION .
VERSION .
KEYWORDS pMGS46 (ARF16-PB1-HA-P2A-OsTIR1)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10247)
AUTHORS Sathyan KM, McKenna BD, Anderson WD, Duarte FM, Core L, Guertin MJ
TITLE An improved auxin-inducible degron system preserves native protein
levels and enables rapid and specific protein depletion.
JOURNAL Genes Dev. 2019 Oct 1;33(19-20):1441-1455. doi:
10.1101/gad.328237.119. Epub 2019 Aug 29.
PUBMED 31467088
REFERENCE 2 (bases 1 to 10247)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..10247
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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 186..208
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 205..221
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 242..260
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
misc_feature 295..1098
/label=HA-L
/note="left homology arm from the adeno-associated virus
integration site (AAVS1) within intron 1 of the human
PPP1R12C gene"
enhancer 1155..1458
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1459..1662
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 1613..1633
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 1703..1723
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2261..2287
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 2297..2353
/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 2363..4090
/codon_start=1
/product="rice transport inhibitor response 1 protein"
/label=OsTIR1
/note="F-box protein that mediates auxin-dependent
interaction of a ubiquitin ligase with an auxin-inducible
degron (Tan et al., 2007)"
/translation="MTYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFV
GNCYAVRAGRVAARFPNVRALTVKGKPHFADFNLVPPDWGGYAGPWIEAAARGCHGLEE
LRMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEV
EDRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTL
AKILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYP
LCAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELR
VFPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRL
CILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSI
AFAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKG
CQVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKIL"
polyA_signal 4264..4385
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4301..4320)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4355..4374
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
promoter 4394..4894
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 4813..4831
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 4916..5515
/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(4916..4935)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 5412..5432
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 5522..5746
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
misc_feature 5749..6585
/label=HA-R
/note="right homology arm from the adeno-associated virus
integration site (AAVS1) within intron 1 of the human
PPP1R12C gene"
primer_bind complement(6619..6638)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(6620..6638)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6645..6662)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(6645..6661)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6654..6676)
/label=M13/pUC Forward
/note="In lacZ gene"
CDS 6799..7101
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind 6980..6999
/label=ccdB-fwd
/note="ccdB gene, forward primer"
CDS 7450..8244
/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
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(7504..7523)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 8114..8133
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
CDS complement(8494..9354)
/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 9117..9136
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 9478..10066
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9967..9986
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 10220..10237
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctca
241 gaattaaccc tcactaaagg gactagtcct gcaggtttaa acgaattcgc cctttgcttt
301 ctctgaccag cattctctcc cctgggcctg tgccgctttc tgtctgcagc ttgtggcctg
361 ggtcacctct acggctggcc cagatccttc cctgccgcct ccttcaggtt ccgtcttcct
421 ccactccctc ttccccttgc tctctgctgt gttgctgccc aaggatgctc tttccggagc
481 acttccttct cggcgctgca ccacgtgatg tcctctgagc ggatcctccc cgtgtctggg
541 tcctctccgg gcatctctcc tccctcaccc aaccccatgc cgtcttcact cgctgggttc
601 ccttttcctt ctccttctgg ggcctgtgcc atctctcgtt tcttaggatg gccttctccg
661 acggatgtct cccttgcgtc ccgcctcccc ttcttgtagg cctgcatcat caccgttttt
721 ctggacaacc ccaaagtacc ccgtctccct ggctttagcc acctctccat cctcttgctt
781 tctttgcctg gacaccccgt tctcctgtgg attcgggtca cctctcactc ctttcatttg
841 ggcagctccc ctacccccct tacctctcta gtctgtgcta gctcttccag ccccctgtca
901 tggcatcttc caggggtccg agagctcagc tagtcttctt cctccaaccc gggcccctat
961 gtccacttca ggacagcatg tttgctgcct ccagggatcc tgtgtccccg agctgggacc
1021 accttatatt cccagggccg gttaatgtgg ctctggttct gggtactttt atctgtcccc
1081 tccaccccac agtggggcaa ttaatagtaa tcaattacgg ggtcattagt tcatagccca
1141 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
1201 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
1261 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
1321 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
1381 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
1441 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
1501 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
1561 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
1621 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
1681 atccgctagc gctgccacca tgcctaagaa gaagcggaag gtgggcagca cactgacttc
1741 cgggcagcgg gatcaccggg acgtcgagaa cgagctgagc tccgccgcct tcagctccca
1801 gagcttcggg gtgcccgaca tgagctttaa gcccggctgc agcagcgacg tggccgtgac
1861 cgacgccggg atgcccagcc aggggctgtg gaacaaccag acccagcgga tgaggacatt
1921 cacaaaggtg cagaagcggg ggagcgtggg ccggagcatc gatatcacca ggtacaggga
1981 ttacgatgag ctgaggcatg atctcgcctg tatgttcggc atccagggcc agctggagga
2041 cccctaccgg atggactgga agctggtgta cgtggaccac gagaacgata tcctgctggt
2101 gggcgatgac ccctgggagg agttcgtggg ctgcgtgaag agcattaaga ttctgtctgc
2161 tgctgaggtg cagcagatga gcctggatgg agatctggga ggagtgccac ctcagactca
2221 ggcctgtagt gctagtgatg atgctaacgc ctggagaggg tatccatatg acgtcccaga
2281 ctatgccgga agcggagcta ctaacttcag cctgctgaag caggctggag acgtggagga
2341 gaaccctgga cctgctgcca ccatgacata ctttcctgaa gaggtcgtcg aacacatttt
2401 tagcttcctg cctgcacaga gagatagaaa cacagtgagc ctggtctgca aagtgtggta
2461 cgagatcgaa cgcctgagcc ggagaggagt gttcgtcggc aactgctatg ctgtgagagc
2521 aggcagggtc gccgctaggt ttccaaatgt gcgcgcactg accgtcaagg ggaaacccca
2581 cttcgccgac tttaacctgg tgccccctga ttggggagga tacgccggcc cttggatcga
2641 ggcagccgct cgcggctgtc atggactgga ggaactgcgc atgaagcgaa tggtggtctc
2701 tgacgaaagt ctggagctgc tggctcggag cttccctagg tttcgcgcac tggtgctgat
2761 ttcttgcgaa ggcttcagca ccgatggact ggcagccgtg gcctcccact gtaagctgct
2821 gcgggagctg gacctccagg agaatgaagt ggaggataga ggccccagat ggctgtcttg
2881 cttcccagac tcatgtacca gcctggtgtc cctgaacttt gcctgcatca aaggcgaagt
2941 gaatgctggg tccctggagc ggctggtctc aagaagcccc aacctgaggt ctctgcggct
3001 gaaccggagc gtgagcgtgg acactctggc taagattctg ctgagaaccc ctaacctgga
3061 ggatctggga accggcaatc tgacagacga tttccagaca gaatcctact ttaaactgac
3121 ttctgccctg gagaagtgta aaatgctgag gagtctgtca ggattctggg atgcttcacc
3181 cgtgtgcctg agctttatct accctctgtg tgcacagctg acaggcctga acctgagcta
3241 tgcaccaacc ctggacgcca gtgatctgac aaagatgatc tcacgctgcg tgaaactcca
3301 gcgactgtgg gtgctggact gtatttccga taaggggctc caggtggtcg ccagctcctg
3361 caaggacctc caggagctga gagtgttccc atctgatttt tacgtggccg gatatagtgc
3421 tgtcactgag gaaggcctgg tggcagtctc actgggatgc ccaaagctga acagcctgct
3481 gtatttctgt catcagatga ctaatgctgc actggtgacc gtcgccaaga actgccctaa
3541 tttcacccga tttcggctgt gtattctgga accaggcaaa cccgacgtgg tcacatccca
3601 gccactggat gaagggtttg gagctatcgt gagagagtgc aagggactcc agaggctgag
3661 catttccggc ctgctgacag acaaagtgtt catgtacatc ggcaagtatg ctaagcagct
3721 ggagatgctg agcattgcat ttgccggaga ctccgataag ggcatgatgc acgtgatgaa
3781 cgggtgtaag aatctgcgaa aactggaaat ccgggacagc cctttcgggg atgccgctct
3841 gctgggaaac tttgccagat acgagacaat gaggagcctg tggatgtcta gttgcaatgt
3901 gactctgaag ggctgtcagg tcctggctag taaaatgcct atgctgaacg tggaagtcat
3961 taatgagcgg gacgggtcta acgaaatgga ggaaaatcat ggcgacctgc caaaggtgga
4021 gaaactgtat gtgtatcgga ccaccgcagg ggcaagagat gatgctccca actttgtgaa
4081 gattctgtga ggatccatcg gatcccgggc ccgtcgacgg taccgcgggc ccgggatcca
4141 ccggggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
4201 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
4261 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
4321 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
4381 tcttaagttg gggttgcgcc ttttccaagg cagccctggg tttgcgcagg gacgcggctg
4441 ctctgggcgt ggttccggga aacgcagcgg cgccgaccct gggtctcgca cattcttcac
4501 gtccgttcgc agcgtcaccc ggatcttcgc cgctaccctt gtgggccccc cggcgacgct
4561 tcctgctccg cccctaagtc gggaaggttc cttgcggttc gcggcgtgcc ggacgtgaca
4621 aacggaagcc gcacgtctca ctagtaccct cgcagacgga cagcgccagg gagcaatggc
4681 agcgcgccga ccgcgatggg ctgtggccaa tagcggctgc tcagcagggc gcgccgagag
4741 cagcggccgg gaaggggcgg tgcgggaggc ggggtgtggg gcggtagtgt gggccctgtt
4801 cctgcccgcg cggtgttccg cattctgcaa gcctccggag cgcacgtcgg cagtcggctc
4861 cctcgttgac cgaatcaccg acctctctcc ccagggggat ccaccggagc ttaccatgac
4921 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
4981 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
5041 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
5101 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
5161 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
5221 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
5281 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
5341 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
5401 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
5461 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaggtac
5521 cctgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
5581 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
5641 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
5701 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggttac tagggacagg
5761 attggtgaca gaaaagcccc atccttaggc ctcctccttc ctagtctcct gatattgggt
5821 ctaaccccca cctcctgtta ggcagattcc ttatctggtg acacaccccc atttcctgga
5881 gccatctctc tccttgccag aacctctaag gtttgcttac gatggagcca gagaggatcc
5941 tgggagggag agcttggcag ggggtgggag ggaagggggg gatgcgtgac ctgcccggtt
6001 ctcagtggcc accctgcgct accctctccc agaacctgag ctgctctgac gcggctgtct
6061 ggtgcgtttc actgatcctg gtgctgcagc ttccttacac ttcccaagag gagaagcagt
6121 ttggaaaaac aaaatcagaa taagttggtc ctgagttcta actttggctc ttcacctttc
6181 tagtccccaa tttatattgt tcctccgtgc gtcagtttta cctgtgagat aaggccagta
6241 gccagccccg tcctggcagg gctgtggtga ggaggggggt gtccgtgtgg aaaactccct
6301 ttgtgagaat ggtgcgtcct aggtgttcac caggtcgtgg ccgcctctac tccctttctc
6361 tttctccatc cttctttcct taaagagtcc ccagtgctat ctgggacata ttcctccgcc
6421 cagagcaggg tcccgcttcc ctaaggccct gctctgggct tctgggtttg agtccttggc
6481 aagcccagga gaggcgctca ggcttccctg tcccccttcc tcgtccacca tctcatgccc
6541 ctggctctcc tgccccttcc ctacaggggt tcctggctct gctctaaggg cgaattcgcg
6601 gccgctaaat tcaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgtttta
6661 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
6721 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
6781 cgcagcctat acgtacggca gtttaaggtt tacacctata aaagagagag ccgttatcgt
6841 ctgtttgtgg atgtacagag tgatattatt gacacgccgg ggcgacggat ggtgatcccc
6901 ctggccagtg cacgtctgct gtcagataaa gtctcccgtg aactttaccc ggtggtgcat
6961 atcggggatg aaagctggcg catgatgacc accgatatgg ccagtgtgcc ggtctccgtt
7021 atcggggaag aagtggctga tctcagccac cgcgaaaatg acatcaaaaa cgccattaac
7081 ctgatgttct ggggaatata aatgtcaggc atgagattat caaaaaggat cttcacctag
7141 atccttttca cgtagaaagc cagtccgcag aaacggtgct gaccccggat gaatgtcagc
7201 tactgggcta tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt
7261 gggcttacat ggcgatagct agactgggcg gttttatgga cagcaagcga accggaattg
7321 ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc
7381 ttgccgccaa ggatctgatg gcgcagggga tcaagctctg atcaagagac aggatgagga
7441 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
7501 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
7561 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
7621 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
7681 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
7741 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
7801 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
7861 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
7921 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
7981 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
8041 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
8101 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
8161 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
8221 cgccttcttg acgagttctt ctgaattatt aacgcttaca atttcctgat gcggtatttt
8281 ctccttacgc atctgtgcgg tatttcacac cgcatcaggt ggcacttttc ggggaaatgt
8341 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
8401 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
8461 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
8521 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
8581 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
8641 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
8701 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
8761 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
8821 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
8881 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
8941 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
9001 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
9061 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
9121 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
9181 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
9241 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
9301 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
9361 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgacca aaatccctta
9421 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
9481 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
9541 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
9601 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
9661 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
9721 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
9781 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
9841 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
9901 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
9961 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
10021 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
10081 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
10141 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
10201 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
