| 质粒名称: | AF20-double-tag |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 108389 |
| 存储实验室: | Gregory Germino |
|
载体骨架基本信息 |
|
| 载体名称: | modified pCI |
| 载体出品公司: | Promega |
| 完整质粒大小: | 19157 bp |
| 载体类型: | 哺乳细胞表达 |
| 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: | PKD1 / PC1 / polycystin 1 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 14481 bp |
| 启动子: | CMV |
| 标签/融合蛋白: |
mcherry-TEV cleavage-V5 (N terminal on insert)
eGFP-TEV cleavage-S15 (C terminal on insert)
|
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: |
AF20-double-tag was a gift from Gregory Germino (Addgene plasmid # 108389 ; http://n2t.net/addgene:108389 ; RRID:Addgene_108389)
|
| 参考文献部分: | A cleavage product of Polycystin-1 is a mitochondrial matrix protein that affects mitochondria morphology and function when heterologously expressed. Lin CC, Kurashige M, Liu Y, Terabayashi T, Ishimoto Y, Wang T, Choudhary V, Hobbs R, Liu LK, Lee PH, Outeda P, Zhou F, Restifo NP, Watnick T, Kawano H, Horie S, Prinz W, Xu H, Menezes LF, Germino GG. Sci Rep. 2018 Feb 9;8(1):2743. doi: 10.1038/s41598-018-20856-6. 10.1038/s41598-018-20856-6 [pii] |
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 AF20-double-tag 19157 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Express human PC1.
ACCESSION .
VERSION .
KEYWORDS AF20-double-tag
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 19157)
AUTHORS Lin CC, Kurashige M, Liu Y, Terabayashi T, Ishimoto Y, Wang T,
Choudhary V, Hobbs R, Liu LK, Lee PH, Outeda P, Zhou F, Restifo NP,
Watnick T, Kawano H, Horie S, Prinz W, Xu H, Menezes LF, Germino GG
TITLE A cleavage product of Polycystin-1 is a mitochondrial matrix protein
that affects mitochondria morphology and function when
heterologously expressed.
JOURNAL Sci Rep. 2018 Feb 9;8(1):2743. doi: 10.1038/s41598-018-20856-6.
PUBMED 29426897
REFERENCE 2 (bases 1 to 19157)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..19157
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 138..517
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 518..721
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 672..692
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 718..742
/label=LNCX
/note="Human CMV promoter, forward primer"
intron 857..989
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
primer_bind 972..991
/label=pMT2-F
/note="Synthetic intron, forward primer"
promoter 1034..1052
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
intron 1083..1655
/label=-beta-globin intron
/note="intron from rabbit beta-globin gene"
primer_bind 1663..1682
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
protein_bind 1713..1737
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 1815..2522
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(1960..1978)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 2228..2247
/label=mCherry-F
/note="mCherry, forward primer"
CDS 2523..2543
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 2544..2585
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 15435..16151
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 15435..16151
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(15480..15501)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(15741..15760)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 16088..16109
/label=EGFP-C
/note="EGFP, forward primer"
CDS 16152..16172
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 16173..16217
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/label=S-Tag
/translation="KETAAAKFERQHMDS"
polyA_signal 16260..16308
/note="synthetic polyadenylation signal"
rep_origin 16545..17000
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(16632..16651)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 16842..16863
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(17067..17086)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 17186..17208
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(17246..17264)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 17332..17436
/gene="bla"
/label=AmpR promoter
CDS 17437..18297
/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 complement(17655..17674)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 18468..19056
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 18957..18976
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
721 tcgtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
1021 aggctagagt acttaatacg actcactata ggctagtctg gatcgatcct gagaacttca
1081 gggtgagttt ggggaccctt gattgttctt tctttttcgc tattgtaaaa ttcatgttat
1141 atggaggggg caaagttttc agggtgttgt ttagaatggg aagatgtccc ttgtatcacc
1201 atggaccctc atgataattt tgtttctttc actttctact ctgttgacaa ccattgtctc
1261 ctcttatttt cttttcattt tctgtaactt tttcgttaaa ctttagcttg catttgtaac
1321 gaatttttaa attcactttt gtttatttgt cagattgtaa gtactttctc taatcacttt
1381 tttttcaagg caatcagggt atattatatt gtacttcagc acagttttag agaacaattg
1441 ttataattaa atgataaggt agaatatttc tgcatataaa ttctggctgg cgtggaaata
1501 ttcttattgg tagaaacaac tacatcctgg tcatcatcct gcctttctct ttatggttac
1561 aatgatatac actgtttgag atgaggataa aatactctga gtccaaaccg ggcccctctg
1621 ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg
1681 tgctgtctca tcattttggc aaagaattcg gggcaacttt gtacaaaaaa gttgatatat
1741 ccaccatgcc gcccgccgcg cccgcccgcc tggcgctggc cctgggcctg ggcctgtggc
1801 tcggggcgct ggcgatggtt agtaagggcg aggaggataa catggccatc atcaaggagt
1861 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
1921 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
1981 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
2041 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
2101 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
2161 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
2221 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
2281 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
2341 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
2401 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
2461 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
2521 aggagaatct ttattttcag ggcggtaagc ctatccctaa ccctctcctc ggtctcgatt
2581 ctacggatat cgggggcccc gggcgcggct gcgggccctg cgagcccccc tgcctctgcg
2641 gcccagcgcc cggcgccgcc tgccgcgtca actgctcggg ccgcgggctg cggacgctcg
2701 gtcccgcgct gcgcatcccc gcggacgcca cagcgctaga cgtctcccac aacctgctcc
2761 gggcgctgga cgttgggctc ctggcgaacc tctcggcgct ggcagagctg gatataagca
2821 acaacaagat ttctacgtta gaagaaggaa tatttgctaa tttatttaat ttaagtgaaa
2881 taaacctgag tgggaacccg tttgagtgtg actgtggcct ggcgtggctg ccgcgatggg
2941 cggaggagca gcaggtgcgg gtggtgcagc ccgaggcagc cacgtgtgct gggcctggct
3001 ccctggctgg ccagcctctg cttggcatcc ccttgctgga cagtggctgt ggtgaggagt
3061 atgtcgcctg cctccctgac aacagctcag gcaccgtggc agcagtgtcc ttttcagctg
3121 cccacgaagg cctgcttcag ccagaggcct gcagcgcctt ctgcttctcc accggccagg
3181 gcctcgcagc cctctcggag cagggctggt gcctgtgtgg ggcggcccag ccctccagtg
3241 cctcctttgc ctgcctgtcc ctctgctccg gccccccgcc acctcctgcc cccacctgta
3301 ggggccccac cctcctccag cacgtcttcc ctgcctcccc aggggccacc ctggtggggc
3361 cccacggacc tctggcctct ggccagctag cagccttcca catcgctgcc ccgctccctg
3421 tcactgccac acgctgggac ttcggagacg gctccgccga ggtggatgcc gctgggccgg
3481 ctgcctcgca tcgctatgtg ctgcctgggc gctatcacgt gacggccgtg ctggccctgg
3541 gggccggctc agccctgctg gggacagacg tgcaggtgga agcggcacct gccgccctgg
3601 agctcgtgtg cccgtcctcg gtgcagagtg acgagagcct tgacctcagc atccagaacc
3661 gcggtggttc aggcctggag gccgcctaca gcatcgtggc cctgggcgag gagccggccc
3721 gagcggtgca cccgctctgc ccctcggaca cggagatctt ccctggcaac gggcactgct
3781 accgcctggt ggtggagaag gcggcctggc tgcaggcgca ggagcagtgt caggcctggg
3841 ccggggccgc cctggcaatg gtggacagtc ccgccgtgca gcgcttcctg gtctctcggg
3901 tcaccaggag cctagacgtg tggatcggct tctcgactgt gcagggggtg gaggtgggcc
3961 cagcgccgca gggcgaggcc ttcagcctgg agagctgcca gaactggctg cccggggagc
4021 cacacccagc cacagccgag cactgcgtcc ggctcgggcc caccgggtgg tgtaacaccg
4081 acctgtgctc agcgccgcac agctacgtct gcgagctgca gcccggaggc ccagtgcagg
4141 atgccgagaa cctcctcgtg ggagcgccca gtggggacct gcagggaccc ctgacgcctc
4201 tggcacagca ggacggcctc tcagccccgc acgagcccgt ggaggtcatg gtattcccgg
4261 gcctgcgtct gagccgtgaa gccttcctca ccacggccga atttgggacc caggagctcc
4321 ggcggcccgc ccagctgcgg ctgcaggtgt accggctcct cagcacagca gggaccccgg
4381 agaacggcag cgagcctgag agcaggtccc cggacaacag gacccagctg gcccccgcgt
4441 gcatgccagg gggacgctgg tgccctggag ccaacatctg cttgccgctg gacgcctcct
4501 gccaccccca ggcctgcgcc aatggctgca cgtcagggcc agggctaccc ggggccccct
4561 atgcgctatg gagagagttc ctcttctccg ttcccgcggg gccccccgcg cagtactcgg
4621 tcaccctcca cggccaggat gtcctcatgc tccctggtga cctcgttggc ttgcagcacg
4681 acgctggccc tggcgccctc ctgcactgct cgccggctcc cggccaccct ggtccccagg
4741 ccccgtacct ctccgccaac gcctcgtcat ggctgcccca cttgccagcc cagctggagg
4801 gcacttgggc ctgccctgcc tgtgccctgc ggctgcttgc agccacggaa cagctcaccg
4861 tgctgctggg cttgaggccc aaccctggac tgcggctgcc tgggcgctat gaggtccggg
4921 cagaggtggg caatggcgtg tccaggcaca acctctcctg cagctttgac gtggtctccc
4981 cagtggctgg gctgcgggtc atctaccctg ccccccgcga cggccgcctc tacgtgccca
5041 ccaacggctc agccttggtg ctccaggtgg actctggtgc caacgccacg gccacggctc
5101 gctggcctgg gggcagtgtc agcgcccgct ttgagaatgt ctgccctgcc ctggtggcca
5161 ccttcgtgcc cggctgcccc tgggagacca acgataccct gttctcagtg gtagcactgc
5221 cgtggctcag tgagggggag cacgtggtgg acgtggtggt ggaaaacagc gccagccggg
5281 ccaacctcag cctgcgggtg acggcggagg agcccatctg tggcctccgc gccacgccca
5341 gccccgaggc ccgtgtactg cagggagtcc tagtgaggta cagccccgtg gtggaggccg
5401 gctcggacat ggtcttccgg tggaccatca acgacaagca gtccctgacc ttccagaacg
5461 tggtcttcaa tgtcatttat cagagcgcgg cggtcttcaa gctctcactg acggcctcca
5521 accacgtgag caacgtcacc gtgaactaca acgtaaccgt ggagcggatg aacaggatgc
5581 agggtctgca ggtctccaca gtgccggccg tgctgtcccc caatgccacg ctggcactga
5641 cggcgggcgt gctggtggac tcggccgtgg aggtggcctt cctgtggacc tttggggatg
5701 gggagcaggc cctccaccag ttccagcctc cgtacaacga gtccttcccg gttccagacc
5761 cctcggtggc ccaggtgctg gtggagcaca atgtcatgca cacctacgct gccccaggtg
5821 agtacctcct gaccgtgctg gcatctaatg ccttcgagaa cctgacgcag caggtgcctg
5881 tgagcgtgcg cgcctccctg ccctccgtgg ctgtgggtgt gagtgacggc gtcctggtgg
5941 ccggccgggc cgtcaccttc tacccgcacc cgctgccctc gcctgggggt gttctttaca
6001 cgtgggactt cggggacggc tcccctgtcc tgacccagag ccagccggct gccaaccaca
6061 cctatgcctc gaggggcacc taccacgtgc gcctggaggt caacaacacg gtgagcggtg
6121 cggcggccca ggcggatgtg cgcgtctttg aggagctccg cggactcagc gtggacatga
6181 gcctggccgt ggagcagggc gcccccgtgg tggtcagcgc cgcggtgcag acgggcgaca
6241 acatcacgtg gaccttcgac atgggggacg gcaccgtgct gtcgggcccg gaggcaacag
6301 tggagcatgt gtacctgcgg gcacagaact gcacagtgac cgtgggtgcg gccagccccg
6361 ccggccacct ggcccggagc ctgcacgtgc tggtcttcgt cctggaggtg ctgcgcgttg
6421 aacccgccgc ctgcatcccc acgcagcctg acgcgcggct cacggcctac gtcaccggga
6481 acccggccca ctacctcttc gactggacct tcggggatgg ctcctccaac acgaccgtgc
6541 gggggtgccc gacggtgaca cacaacttca cgcggagcgg cacgttcccc ctggcgctgg
6601 tgctgtccag ccgcgtgaac agggcgcatt acttcaccag catctgcgtg gagccagagg
6661 tgggcaacgt caccctgcag ccagagaggc agtttgtgca gctcggggac gaggcctggc
6721 tggtggcatg tgcctggccc ccgttcccct accgctacac ctgggacttt ggcaccgagg
6781 aagccgcccc cacccgtgcc aggggccctg aggtgacgtt catctaccga gacccaggct
6841 cctatcttgt gacagtcacc gcgtccaaca acatctctgc tgccaatgac tcagccctgg
6901 tggaggtgca ggagcccgtg ctggtcacca gcatcaaggt caatggctcc cttgggctgg
6961 agctgcagca gccgtacctg ttctctgctg tgggccgtgg gcgccccgcc agctacctgt
7021 gggatctggg ggacggtggg tggctcgagg gtccggaggt cacccacgct tacaacagca
7081 caggtgactt caccgttagg gtggccggct ggaatgaggt gagccgcagc gaggcctggc
7141 tcaatgtgac ggtgaagcgg cgcgtgcggg ggctcgtcgt caatgcaagc cgcacggtgg
7201 tgcccctgaa tgggagcgtg agcttcagca cgtcgctgga ggccggcagt gatgtgcgct
7261 attcctgggt gctctgtgac cgctgcacgc ccatccctgg gggtcctacc atctcttaca
7321 ccttccgctc cgtgggcacc ttcaatatca tcgtcacggc tgagaacgag gtgggctccg
7381 cccaggacag catcttcgtc tatgtcctgc agctcataga ggggctgcag gtggtgggcg
7441 gtggccgcta cttccccacc aaccacacgg tacagctgca ggccgtggtt agggatggca
7501 ccaacgtctc ctacagctgg actgcctgga gggacagggg cccggccctg gccggcagcg
7561 gcaaaggctt ctcgctcacc gtgctcgagg ccggcaccta ccatgtgcag ctgcgggcca
7621 ccaacatgct gggcagcgcc tgggccgact gcaccatgga cttcgtggag cctgtggggt
7681 ggctgatggt ggccgcctcc ccgaacccag ctgccgtcaa cacaagcgtc accctcagtg
7741 ccgagctggc tggtggcagt ggtgtcgtat acacttggtc cttggaggag gggctgagct
7801 gggagacctc cgagccattt accacccata gcttccccac acccggcctg cacttggtca
7861 ccatgacggc agggaacccg ctgggctcag ccaacgccac cgtggaagtg gatgtgcagg
7921 tgcctgtgag tggcctcagc atcagggcca gcgagcccgg aggcagcttc gtggcggccg
7981 ggtcctctgt gcccttttgg gggcagctgg ccacgggcac caatgtgagc tggtgctggg
8041 ctgtgcccgg cggcagcagc aagcgtggcc ctcatgtcac catggtcttc ccggatgctg
8101 gcaccttctc catccggctc aatgcctcca acgcagtcag ctgggtctca gccacgtaca
8161 acctcacggc ggaggagccc atcgtgggcc tggtgctgtg ggccagcagc aaggtggtgg
8221 cgcccgggca gctggtccat tttcagatcc tgctggctgc cggctcagct gtcaccttcc
8281 gcctgcaggt cggcggggcc aaccccgagg tgctccccgg gccccgtttc tcccacagct
8341 tcccccgcgt cggagaccac gtggtgagcg tgcggggcaa aaaccacgtg agctgggccc
8401 aggcgcaggt gcgcatcgtg gtgctggagg ccgtgagtgg gctgcaggtg cccaactgct
8461 gcgagcctgg catcgccacg ggcactgaga ggaacttcac agcccgcgtg cagcgcggct
8521 ctcgggtcgc ctacgcctgg tacttctcgc tgcagaaggt ccagggcgac tcgctggtca
8581 tcctgtcggg ccgcgacgtc acctacacgc ccgtggccgc ggggctgttg gagatccagg
8641 tgcgcgcctt caacgccctg ggcagtgaga accgcacgct ggtgctggag gttcaggacg
8701 ccgtccagta tgtggccctg cagagcggcc cctgcttcac caaccgctcg gcgcagtttg
8761 aggccgccac cagccccagc ccccggcgtg tggcctacca ctgggacttt ggggatgggt
8821 cgccagggca ggacacagat gagcccaggg ccgagcactc ctacctgagg cctggggact
8881 accgcgtgca ggtgaacgcc tccaacctgg tgagcttctt cgtggcgcag gccacggtga
8941 ccgtccaggt gctggcctgc cgggagccgg aggtggacgt ggtcctgccc ctgcaggtgc
9001 tgatgcggcg atcacagcgc aactacttgg aggcccacgt tgacctgcgc gactgcgtca
9061 cctaccagac tgagtaccgc tgggaggtgt atcgcaccgc cagctgccag cggccggggc
9121 gcccagcgcg tgtggccctg cccggcgtgg acgtgagccg gcctcggctg gtgctgccgc
9181 ggctggcgct gcctgtgggg cactactgct ttgtgtttgt cgtgtcattt ggggacacgc
9241 cactgacaca gagcatccag gccaatgtga cggtggcccc cgagcgcctg gtgcccatca
9301 ttgagggtgg ctcataccgc gtgtggtcag acacacggga cctggtgctg gatgggagcg
9361 agtcctacga ccccaacctg gaggacggcg accagacgcc gctcagtttc cactgggcct
9421 gtgtggcttc gacacagagg gaggctggcg ggtgtgcgct gaactttggg ccccgcggga
9481 gcagcacggt caccattcca cgggagcggc tggcggctgg cgtggagtac accttcagcc
9541 tgaccgtgtg gaaggccggc cgcaaggagg aggccaccaa ccagacggtg ctgatccgga
9601 gtggccgggt gcccattgtg tccttggagt gtgtgtcctg caaggcacag gccgtgtacg
9661 aagtgagccg cagctcctac gtgtacttgg agggccgctg cctcaattgc agcagcggct
9721 ccaagcgagg gcggtgggct gcacgtacgt tcagcaacaa gacgctggtg ctggatgaga
9781 ccaccacatc cacgggcagt gcaggcatgc gactggtgct gcggcggggc gtgctgcggg
9841 acggcgaggg atacaccttc acgctcacgg tgctgggccg ctctggcgag gaggagggct
9901 gcgcctccat ccgcctgtcc cccaaccgcc cgccgctggg gggctcttgc cgcctcttcc
9961 cactgggcgc tgtgcacgcc ctcaccacca aggtgcactt cgaatgcacg ggctggcatg
10021 acgcggagga tgctggcgcc ccgctggtgt acgccctgct gctgcggcgc tgtcgccagg
10081 gccactgcga ggagttctgt gtctacaagg gcagcctctc cagctacgga gccgtgctgc
10141 ccccgggttt caggccacac ttcgaggtgg gcctggccgt ggtggtgcag gaccagctgg
10201 gagccgctgt ggtcgccctc aacaggtctt tggccatcac cctcccagag cccaacggca
10261 gcgcaacggg gctcacagtc tggctgcacg ggctcaccgc tagtgtgctc ccagggctgc
10321 tgcggcaggc cgatccccag cacgtcatcg agtactcgtt ggccctggtc accgtgctga
10381 acgagtacga gcgggccctg gacgtggcgg cagagcccaa gcacgagcgg cagcaccgag
10441 cccagatacg caagaacatc acggagactc tggtgtccct gagggtccac actgtggatg
10501 acatccagca gatcgctgct gcgctggccc agtgcatggg gcccagcagg gagctcgtat
10561 gccgctcgtg cctgaagcag acgctgcaca agctggaggc catgatgctc atcctgcagg
10621 cagagaccac cgcgggcacc gtgacgccca ccgccatcgg agacagcatc ctcaacatca
10681 caggagacct catccacctg gccagctcgg acgtgcgggc accacagccc tcagagctgg
10741 gagccgagtc accatctcgg atggtggcgt cccaggccta caacctgacc tctgccctca
10801 tgcgcatcct catgcgctcc cgcgtgctca acgaggagcc cctgacgctg gcgggcgagg
10861 agatcgtggc ccagggcaag cgctcggacc cgcggagcct gctgtgctat ggcggcgccc
10921 cagggcctgg ctgccacttc tccatccccg aggctttcag cggggccctg gccaacctca
10981 gtgacgtggt gcagctcatc tttctggtgg actccaatcc ctttcccttt ggctatatca
11041 gcaactacac cgtctccacc aaggtggcct cgatggcatt ccagacacag gccggcgccc
11101 agatccccat cgagcggctg gcctcagagc gcgccatcac cgtgaaggtg cccaacaact
11161 cggactgggc tgcccggggc caccgcagct ccgccaactc cgccaactcc gttgtggtcc
11221 agccccaggc ctccgtcggt gctgtggtca ccctggacag cagcaaccct gcggccgggc
11281 tgcatctgca gctcaactat acgctgctgg acggccacta cctgtctgag gaacctgagc
11341 cctacctggc agtctaccta cactcggagc cccggcccaa tgagcacaac tgctcggcta
11401 gcaggaggat ccgcccagag tcactccagg gtgctgacca ccggccctac accttcttca
11461 tttccccggg gagcagagac ccagcgggga gttaccatct gaacctctcc agccacttcc
11521 gctggtcggc gctgcaggtg tccgtgggcc tgtacacgtc cctgtgccag tacttcagcg
11581 aggaggacat ggtgtggcgg acagaggggc tgctgcccct ggaggagacc tcgccccgcc
11641 aggccgtctg cctcacccgc cacctcaccg ccttcggcgc cagcctcttc gtgcccccaa
11701 gccatgtccg ctttgtgttt cctgagccga cagcggatgt aaactacatc gtcatgctga
11761 catgtgctgt gtgcctggtg acctacatgg tcatggccgc catcctgcac aagctggacc
11821 agttggatgc cagccggggc cgcgccatcc ctttctgtgg gcagcggggc cgcttcaagt
11881 acgagatcct cgtcaagaca ggctggggcc ggggctcagg taccacggcc cacgtgggca
11941 tcatgctgta tggggtggac agccggagcg gccaccggca cctggacggc gacagagcct
12001 tccaccgcaa cagcctggac atcttccgga tcgccacccc gcacagcctg ggtagcgtgt
12061 ggaagatccg agtgtggcac gacaacaaag ggctcagccc tgcctggttc ctgcagcacg
12121 tcatcgtcag ggacctgcag acggcacgca gcgccttctt cctggtcaat gactggcttt
12181 cggtggagac ggaggccaac gggggcctgg tggagaagga ggtgctggcc gcgagcgacg
12241 cagccctttt gcgcttccgg cgcctgctgg tggctgagct gcagcgtggc ttctttgaca
12301 agcacatctg gctctccata tgggaccggc cgcctcgtag ccgtttcact cgcatccaga
12361 gggccacctg ctgcgttctc ctcatctgcc tcttcctggg cgccaacgcc gtgtggtacg
12421 gggctgttgg cgactctgcc tacagcacgg ggcatgtgtc caggctgagc ccgctgagcg
12481 tcgacacagt cgctgttggc ctggtgtcca gcgtggttgt ctatcccgtc tacctggcca
12541 tcctttttct cttccggatg tcccggagca aggtggctgg gagcccgagc cccacacctg
12601 ccgggcagca ggtgctggac atcgacagct gcctggactc gtccgtgctg gacagctcct
12661 tcctcacgtt ctcaggcctc cacgctgagg cctttgttgg acagatgaag agtgacttgt
12721 ttctggatga ttctaagagt ctggtgtgct ggccctccgg cgagggaacg ctcagttggc
12781 cggacctgct cagtgacccg tccattgtgg gtagcaatct gcggcagctg gcacggggcc
12841 aggcgggcca tgggctgggc ccagaggagg acggcttctc cctggccagc ccctactcgc
12901 ctgccaaatc cttctcagca tcagatgaag acctgatcca gcaggtcctt gccgaggggg
12961 tcagcagccc agcccctacc caagacaccc acatggaaac ggacctgctc agcagcctgt
13021 ccagcactcc tggggagaag acagagacgc tggcgctgca gaggctgggg gagctggggc
13081 cacccagccc aggcctgaac tgggaacagc cccaggcagc gaggctgtcc aggacaggac
13141 tggtggaggg tctgcggaag cgcctgctgc cggcctggtg tgcctccctg gcccacgggc
13201 tcagcctgct cctggtggct gtggctgtgg ctgtctcagg gtgggtgggt gcgagcttcc
13261 ccccgggcgt gagtgttgcg tggctcctgt ccagcagcgc cagcttcctg gcctcattcc
13321 tcggctggga gccactgaag gtcttgctgg aagccctgta cttctcactg gtggccaagc
13381 ggctgcaccc ggatgaagat gacaccctgg tagagagccc ggctgtgacg cctgtgagcg
13441 cacgtgtgcc ccgcgtacgg ccaccccacg gctttgcact cttcctggcc aaggaagaag
13501 cccgcaaggt caagaggcta catggcatgc tgcggagcct cctggtgtac atgctttttc
13561 tgctggtgac cctgctggcc agctatgggg atgcctcatg ccatgggcac gcctaccgtc
13621 tgcaaagcgc catcaagcag gagctgcaca gccgggcctt cctggccatc acgcggtctg
13681 aggagctctg gccatggatg gcccacgtgc tgctgcccta cgtccacggg aaccagtcca
13741 gcccagagct ggggccccca cggctgcggc aggtgcggct gcaggaagca ctctacccag
13801 accctcccgg ccccagggtc cacacgtgct cggccgcagg aggcttcagc accagcgatt
13861 acgacgttgg ctgggagagt cctcacaatg gctcggggac gtgggcctat tcagcgccgg
13921 atctgctggg ggcatggtcc tggggctcct gtgccgtgta tgacagcggg ggctacgtgc
13981 aggagctggg cctgagcctg gaggagagcc gcgaccggct gcgcttcctg cagctgcaca
14041 actggctgga caacaggagc cgcgctgtgt tcctggagct cacgcgctac agcccggccg
14101 tggggctgca cgccgccgtc acgctgcgcc tcgagttccc ggcggccggc cgcgccctgg
14161 ccgccctcag cgtccgcccc tttgcgctgc gccgcctcag cgcaggcctc tcgctgcctc
14221 tgctcacctc ggtgtgcctg ctgctgttcg ccgtgcactt cgccgtggcc gaggcccgta
14281 cttggcacag ggaagggcgc tggcgcgtgc tgcggctcgg agcctgggcg cggtggctgc
14341 tggtggcgct gacggcggcc acggcactgg tacgcctcgc ccagctgggt gccgctgacc
14401 gccagtggac ccgtttcgtg cgcggccgcc cgcgccgctt cactagcttc gaccaggtgg
14461 cgcagctgag ctccgcagcc cgtggcctgg cggcctcgct gctcttcctg cttttggtca
14521 aggctgccca gcagctacgc ttcgtgcgcc agtggtccgt ctttggcaag acattatgcc
14581 gagctctgcc agagctcctg ggggtcacct tgggcctggt ggtgctcggg gtagcctacg
14641 cccagctggc catcctgctc gtgtcttcct gtgtggactc cctctggagc gtggcccagg
14701 ccctgttggt gctgtgccct gggactgggc tctctaccct gtgtcctgcc gagtcctggc
14761 acctgtcacc cctgctgtgt gtggggctct gggcactgcg gctgtggggc gccctacggc
14821 tgggggctgt tattctccgc tggcgctacc acgccttgcg tggagagctg taccggccgg
14881 cctgggagcc ccaggactac gagatggtgg agttgttcct gcgcaggctg cgcctctgga
14941 tgggcctcag caaggtcaag gagttccgcc acaaagtccg ctttgaaggg atggagccgc
15001 tgccctctcg ctcctccagg ggctccaagg tatccccgga tgtgccccca cccagcgctg
15061 gctccgatgc ctcgcacccc tccacctcct ccagccagct ggatgggctg agcgtgagcc
15121 tgggccggct ggggacaagg tgtgagcctg agccctcccg cctccaagcc gtgttcgagg
15181 ccctgctcac ccagtttgac cgactcaacc aggccacaga ggacgtctac cagctggagc
15241 agcagctgca cagcctgcaa ggccgcagga gcagccgggc gcccgccgga tcttcccgtg
15301 gcccatcccc gggcctgcgg ccagcactgc ccagccgcct tgcccgggcc agtcggggtg
15361 tggacctggc cactggcccc agcaggacac cccttcgggc caagaacaag gtccacccca
15421 gcagcactaa gcttatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
15481 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
15541 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
15601 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
15661 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
15721 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
15781 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
15841 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
15901 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
15961 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
16021 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
16081 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
16141 agctgtacaa ggagaatctt tattttcagg gcaaagaaac cgctgctgct aaattcgaac
16201 gccagcacat ggacagctga taatagattt aaatcaactt tgttgtacaa agttgcccca
16261 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtggt taacaacaac
16321 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag
16381 taaaacctct acaaatgtgg taaaatcgat aaggatcaat tgcctttttt tttaattttt
16441 attttatttt atttttgaga tggccagatc cgggctggcg taatagcgaa gaggcccgca
16501 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg
16561 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
16621 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
16681 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
16741 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
16801 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
16861 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
16921 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
16981 aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
17041 tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
17101 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
17161 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
17221 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
17281 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
17341 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
17401 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
17461 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
17521 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
17581 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
17641 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
17701 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
17761 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
17821 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
17881 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
17941 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
18001 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
18061 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
18121 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
18181 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
18241 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
18301 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
18361 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
18421 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
18481 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
18541 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
18601 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
18661 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
18721 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
18781 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
18841 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
18901 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
18961 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
19021 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
19081 cggttcctgg ccttttgctg gccttttgct cacatggctc gacagatcga tctggccaaa
19141 aaggccggac cggatct
//
