| 質粒名稱: | pAR18_StrepI_mFASm_H8_pET22b |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 122847 |
| 存儲實驗室: | Martin Grininger |
|
載體骨架基本信息 |
|
| 載體名稱: | pET22b |
| 載體出品公司: | Novagen/Merck Millipore |
| 空載體大小: | 5310 bp |
| 完整質粒大小: | 12987bp |
| 載體類型: | 細菌表達 |
| 篩選標記: | 新霉素（G418） |
| 載體抗性: | 氨芐霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷貝數: | 低拷貝 |
|
插入基因信息 |
|
| 插入基因名稱: | mouse FASN/ mouse fatty acid synthase |
| 物種來源: | M. musculus (mouse) |
| 插入大小: | 7647bp |
| 基因突變: | wildtype |
| 啟動子: | T7 promoter |
| 標簽/融合蛋白: |
Twin-Strep tag (N terminal on insert)
His8 tag (C terminal on insert)
|
| 克隆方法: | 限制性內切酶 |
| 5’測序引物 | T7 prom |
| 3’測序引物 | T7 term |
|
文獻引用方法 |
|
| 材料和方法部分: | pAR18_StrepI_mFASm_H8_pET22b was a gift from Martin Grininger (Addgene plasmid # 122847 ; //n2t.net/addgene:122847 ; RRID:Addgene_122847) |
| 參考文獻部分: | Probing the modularity of megasynthases by rational engineering of a fatty acid synthase Type I. Rittner A, Paithankar KS, Drexler DJ, Himmler A, Grininger M. Protein Sci. 2019 Feb;28(2):414-428. doi: 10.1002/pro.3550. Epub 2018 Dec 20. 10.1002/pro.3550 PubMed 30394635 |
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pAR18_StrepI_mFASm_H8_pET22b載體質粒基本信息
pAR18_StrepI_mFASm_H8_pET22b質粒圖譜載體圖譜和 pAR18_StrepI_mFASm_H8_pET22b載體序列質粒序列多克隆位點信息
pAR18_StrepI_mFASm_H8_pET22b質粒載體簡介
pAR18_StrepI_mFASm_H8_pET22b質粒序列載體序列
LOCUS pAR18_StrepI_mFASm_H8_pET22b 12983 bp DNA circular SYN 14-APR-2025
DEFINITION Expresses murine type I fatty acid synthase (FASN) in Escherichia
coli. N-terminal Twin-Strep tag; C-terminal H8-tag; in pET22b
vector..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12983)
AUTHORS Rittner A, Paithankar KS, Drexler DJ, Himmler A, Grininger M
TITLE Probing the modularity of megasynthases by rational engineering of a
fatty acid synthase Type I.
JOURNAL Protein Sci. 2019 Feb;28(2):414-428. doi: 10.1002/pro.3550. Epub
2018 Dec 20.
PUBMED 30394635
REFERENCE 2 (bases 1 to 12983)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
//gt12365.cn
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1002/pro.3550";
journalName: "Protein Sci"; date: "2019-02"; volume: "28"; issue:
"2"; pages: "414-428"
COMMENT Sequence Label: pAR18_StrepI_mFASm_H8_pET22b
FEATURES Location/Qualifiers
source 1..12983
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 154..173
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 257..276
/label=T7
/note="T7 promoter, forward primer"
promoter 257..275
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 276..300
/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)."
RBS 315..337
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS join(348..353,354..377,378..413,414..437)
/codon_start=1
/product="two Strep-Tag(R)II moieties connected by a linker
for enhanced binding to Strep-Tactin(R), an engineered form
of streptavidin (Schmidt et al., 2013)"
/label=Twin-Strep-tag
/translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
CDS 7965..7988
/codon_start=1
/product="8xHis affinity tag"
/label=8xHis
/translation="HHHHHHHH"
primer_bind complement(8041..8059)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 8055..8102
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 8139..8594
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(8226..8245)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 8436..8457
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 8620..8724
/gene="bla"
/label=AmpR promoter
CDS join(8725..8793,8794..9585)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(8943..8962)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 9756..10344
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 10245..10264
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 10498..10515
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 10530..10669
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(10595..10614)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 10755..10777
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(10771..10962)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(11771..12853)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 12815..12834
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(12854..12931)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
61 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
121 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
181 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat
241 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
301 cctctagaaa taattttgtt taactttaag aaggagatat acatatgagc gcttggagcc
361 atccacaatt tgagaagggt ggaggttctg gcggtggatc gggaggttca gcgtggagcc
421 acccgcagtt cgaaaaaggc gccggatccg aggaggtggt gatagccggt atgtcgggga
481 agttgcccga gtcagagaac ctacaggagt tctgggccaa cctcattggt ggtgtggaca
541 tggtcacaga tgatgacagg agatggaagg ctgggctcta tggattaccc aagcggtctg
601 gaaagctgaa ggatctctcc aagttcgacg cctccttttt tggggtccac cccaagcagg
661 cacacacaat ggacccccag cttcggctgc tgttggaagt cagctatgaa gcaattgtgg
721 atggaggtat caacccagcc tcactccgag gaacgaacac tggcgtctgg gtgggtgtga
781 gtggttcaga ggcatccgag gcccttagca gagatcccga gacgcttctg ggctacagca
841 tggtgggctg ccagcgtgca atgatggcca accggctctc tttcttcttc gacttcaaag
901 gaccaagcat tgccctggac acagcctgct cctccagctt gctggcacta cagaatgcct
961 accaggccat ccgtagtggg gaatgccccg cggcccttgt gggtgggatc aacctgctcc
1021 tgaagccgaa cacctctgtg cagttcatga agctgggcat gctcagcccg gacggcacct
1081 gcagatcctt tgatgattca gggagtggat attgtcgctc tgaggctgtt gtagcagttc
1141 tgctgactaa gaagtccctg gctcggcggg tctatgccac gattctgaat gccggcacca
1201 atacagatgg cagcaaggag caaggtgtaa cattcccctc tggagaagtc caagaacaac
1261 tcatctgctc tctgtatcag ccagctggtc tggccccgga gtcgcttgag tatattgaag
1321 cccatggcac gggcaccaag gtgggtgacc cccaggaact gaatggcatt actcggtccc
1381 tgtgcgcctt ccgccaggcc cctctgttaa ttggctccac caaatccaac atgggacacc
1441 ctgagcctgc ctctgggctt gcagccctga ccaaggtgct gttatccctg gagcatgggg
1501 tctgggcccc taacctgcac ttccacaacc ccaaccctga gatcccagca cttcttgatg
1561 ggcggctgca ggtggtcgat aggcccctgc ctgttcgtgg tggcaacgtg ggcatcaact
1621 catttggctt cggaggctcc aatgttcatg tcatcctcca gcccaacaca cggcaggccc
1681 ctgcgcccac tgcacacgct gcccttcccc atttgctgca cgccagtgga cgcaccttag
1741 aggcagtgca ggacctgctg gaacagggcc gccagcacag ccaggacctg gcctttgtga
1801 gcatgctcaa tgacattgcg gcaaccccta cagcagccat gcccttcagg ggttacactg
1861 tgctaggtgt tgagggccgt gtccaagaag tgcagcaagt gtccaccaac aagcgcccac
1921 tctggttcat ctgctcaggg atgggcacgc agtggcgcgg gatggggctg agcctcatgc
1981 gcctggacag cttccgtgag tctatcctgc gctccgatga ggctgtgaag ccgttgggag
2041 tgaaagtgtc agatctgctg ttgagcacag atgagcgcac ctttgatgac atcgtgcatg
2101 cctttgtgag cctcactgcc atccagattg ccctcatcga cctactgact tctgtgggac
2161 tgaaacctga cggcatcatt gggcactcct tgggagaggt tgcctgtggc tatgcagatg
2221 gctgtctctc ccagagagag gctgtgcttg cagcttactg gcgaggccag tgcatcaaag
2281 atgcccacct cccgcctgga tccatggcag ctgttggttt gtcctgggag gaatgtaaac
2341 agcgctgccc cgctggcgtg gtgcctgcct gccacaactc tgaggacacc gtgaccatct
2401 ctggacctca ggctgcagtg aatgaatttg tggagcagct aaagcaagaa ggtgtgtttg
2461 ccaaggaggt acgaacagga ggcctggctt tccactccta cttcatggaa ggaattgccc
2521 ccacattgct gcaggctctc aagaaggtga tccgggaacc acggccgcgc tcggctcgat
2581 ggctcagcac ctctatccct gaggcccagt ggcagagcag cctggcccgc acatcttctg
2641 ccgagtacaa tgtcaacaac ctggtgagcc ctgtgctctt ccaggaagca ctgtggcaca
2701 tccctgagca tgccgtggtg ctggagattg cgccccacgc actgttgcag gctgtcctga
2761 agcgaggcgt gaagtccagc tgcaccatca ttcccttgat gaagagggat cataaagata
2821 acttggagtt ctttctcacc aaccttggca aggtgcacct cacaggcatc aatgtcaacc
2881 ctaacgcctt gttcccacct gtggagttcc cggctccccg agggactcct ctcatctccc
2941 ctcacatcaa gtgggaccac agtcagactt gggatgtccc ggttgctgag gacttcccaa
3001 acggctccag ctcctcctct gctacagtct acagcatcga cgccagtcct gagtcgcccg
3061 accactacct ggtagaccac tgcattgacg gccgggtcat cttccctggc actggctacc
3121 tgtgcctggt gtggaagaca ctggctcgca gcctgggctt gtccctagaa gagacccctg
3181 tggtatttga gaatgtgtcg tttcatcagg ccactatact acccaagaca ggaaccgtgg
3241 cgctggaggt gaggctgcta gaggcctccc atgcctttga ggtgtctgac actggcaatc
3301 tgattgtgag cggaaaagtg tacctgtggg aagacccgaa ctccaagtta ttcgaccacc
3361 cagaagtccc aacaccccct gagtctgcat cggtctcccg cctgacccag ggagaagtat
3421 acaaggagct gcggctgcgt ggctatgatt atggccctca gttccagggc atctgtgagg
3481 ccacccttga aggtgaacaa ggcaagctgc tctggaaaga taactgggtg accttcatgg
3541 acacaatgct gcaggtatcc attctgggtt ctagccagca gagtctacag ctacctaccc
3601 gtgtgaccgc catctatatc gaccctgcca cccaccgtca gaaggtgtac aggctgaagg
3661 aggacactca agtggctgat gtgacaacga gccgctgtct gggcataacg gtctctggtg
3721 gtatccacat ctcaagacta cagacgacag caacctcacg gcggcagcaa gaacagctgg
3781 tccccacctt ggaaaagttc gttttcacac cgcacatgga ggctgagtgc ctgtctgaga
3841 gcactgccct gcagaaggag ctgcaactgt gcaagggtct ggcacgggct ctgcagacca
3901 aggccaccca gcaagggctg aaggcggcaa tgcttgggca agaggaccct ccacagcacg
3961 ggctgcctcg actcctggca gctgcttgcc agttgcagct caacgggaac ctgcagctgg
4021 agctgggaga agcgctggct caagagaggc tcctgctgcc agaagaccct ctgatcagtg
4081 gcctcctcaa ctcccaggcc ctcaaggcct gcgtagacac agccctggag aacttgtcta
4141 ctctcaagat gaaggtggca gaggtgctgg ctggagaagg ccacttgtat tcccgaatcc
4201 cggcactgct caacacccag cccatgctac aactggaata cacagccacc gaccggcacc
4261 cccaggccct gaaggatgtt cagaccaaac tgcagcagca tgatgtggcg cagggccagt
4321 ggaacccttc cgaccctgcg cccagcagcc tgggtgccct tgaccttctg gtgtgcaact
4381 gtgcattagc caccctgggg gatccagcct tggccctgga caacatggta gctgccctca
4441 aggaaggtgg tttcctgcta gtgcacacag tgctcaaagg acatgccctt ggggagaccc
4501 tggcctgcct accctctgag gtgcagcctg cgcccagcct cctaagccag gaggagtggg
4561 agagcctgtt ctcgaggaag gcactacacc tggtgggcct taaaaggtcc ttctacggta
4621 ctgcgctgtt cctgtgccgg cgagccatcc cacaggagaa acctatcttc ctgtctgtgg
4681 aggataccag cttccagtgg gtggactctc tgaagagcac tctggccacg tcctcctccc
4741 agcctgtgtg gctaacggcc atggactgcc ccacctcggg tgtggtgggt ttggtgaatt
4801 gtctccgaaa agagccgggt ggacaccgga ttcggtgtat cctgctgtcc aacctcagca
4861 acacatctca cgcccccaag ttggaccctg gctctccaga gctacagcag gtgctaaagc
4921 atgacctcgt gatgaacgtg taccgggacg gggcctgggg tgccttccgt cacttccagt
4981 tagagcagga caagcccaag gagcagacag cgcatgcctt tgtaaacgtc ctcacccgag
5041 gggacctcgc ctccatccgc tgggtctcct cccccctgaa gcacacgcag ccctcgagct
5101 caggagcaca gctctgcact gtctactacg cctcactgaa cttccgagac atcatgctgg
5161 ccacgggcaa gctgtcccct gatgccattc caggtaaatg ggccagccga gactgcatgc
5221 tcggcatgga gttctcaggc cgggataggt gtggccggcg tgtgatgggg ctggttcctg
5281 cagaaggcct ggccacctca gtcctgctat catctgactt cctctgggat gtaccctcca
5341 gctggaccct ggaggaggcg gcctctgtgc ccgtcgtcta taccactgct tactactcgt
5401 tagtggttcg cgggcgcatc cagcgtgggg agaccgtgct catccactca ggttcaggtg
5461 gtgtgggcca agcggccatt tccattgccc tcagtctggg ctgccgcgtc ttcaccactg
5521 tgggctctgc agagaagcga gcatacctcc aggccaggtt ccctcagctt gatgacacca
5581 gctttgccaa ctcgagggac acatcatttg agcagcacgt gttactgcac acaggtggca
5641 aaggggtcga cctggtcctc aactcactgg cagaagagaa gctgcaggcc agtgtgcggt
5701 gcttggctca gcatggtcgc ttcttagaga ttggcaaatt tgatctttct aacaaccacc
5761 ctctgggcat ggctatcttc ttgaagaacg tcactttcca tgggatcctg ctggacgccc
5821 tttttgagga ggccaatgac agctggcggg aggtggcggc actcctgaag gctggcattc
5881 gtgatggagt cgtgaagccc ctcaagtgca cagtgtttcc caaggcccag gtggaagatg
5941 ccttccgcta catggctcag gggaaacaca ttggcaaagt ccttgtccag gtacgggagg
6001 aggagcctga ggctgtgctg ccaggggctc agcccaccct gatttctgcc atctccaaga
6061 ccttctgccc agcccataag agttacatca tcactggtgg cctaggtggc tttggcctgg
6121 agctggcccg gtggctcgtg cttcgcggag cccagaggct tgtgctgact tcccgatctg
6181 gaatccgcac cggctaccaa gccaagcaca ttcgggagtg gagacgccag ggcatccaag
6241 tgctcgtgtc aacaagcaac gtgagctcac tggagggggc ccgtgctctc atcgccgaag
6301 ccacaaagct ggggcccgtt gggggtgtct tcaacctggc catggttttg agggatgcca
6361 tgctggagaa ccagacccca gagctcttcc aggatgtcaa caagcccaaa tacaatggca
6421 ccctgaacct tgacagggca acccgggaag cctgccctga gctggactac tttgtggcct
6481 tctcctctgt aagctgcggg cgtggtaatg ctggccaaac taactacggc ttcgccaact
6541 ctaccatgga gcgtatatgt gaacagcgca ggcacgatgg cctcccaggc cttgccgtgc
6601 agtggggtgc cattggtgac gtgggcattg tcctggaagc gatgggcacc aatgacacag
6661 tcatcggagg tacgctgcct cagcgcatct cctcctgcat ggaggtactg gacctcttcc
6721 tgaatcagcc ccacgcagtc ctgagcagct ttgtgctggc agagaagaaa gctgtggccc
6781 atggggacgg ggacacccag agggatctgg tgaaagctgt agcacacatc ctaggcatcc
6841 gagacctcgc aggtattaac ctggacagca cgctggcaga cctcggcctg gactcgctca
6901 tgggtgtgga agttcgtcag atcctggaac gagaacacga tctggtgctg cccatgcgtg
6961 aggtgcggca gctcacgctg cggaaacttc aggaaatgtc ctccaagact gactcggcta
7021 ctgacacgac agcccccaag tccaggagtg acacgtctct gaagcagaac caactgaacc
7081 tgagcacact gctggtgaac cctgagggtc ctaccctaac ccagctcaac tcggtgcaga
7141 gctctgagcg gcctctgttc cttgtgcacc ccattgaggg ttccaccacc gtgttccaca
7201 gtctggctgc caagctcagt gtgcccacct acggcctgca gtgcacccaa gctgcccccc
7261 tggatagcat tccgaacctg gctgcctact acatagattg catcaagcaa gtgcagcctg
7321 agggacccta ccgcatagct gggtactcat ttggagcctg tgtagccttc gagatgtgct
7381 cccagctgca ggcccagcag ggcccagccc cgacccacaa caacctcttc ctgtttgacg
7441 gctcacacac ctacgtgttg gcctacaccc agagctaccg ggcaaagatg accccaggct
7501 gtgaagccga ggccgaggct gaggccttat gcttcttcat aaagcagttt cttgatgtgg
7561 aacacagcaa ggtgctggag gccctgctgc cactgaagag cctggaagat cgggtggctg
7621 cctccgtgga ccttatcact aagagtcacc acagcctgga ccgccgagag ctgagctttg
7681 ctgccgtgtc cttctaccac aagctccggg cagctgatca gtataagccc aaggccaagt
7741 accatggcaa cgtgacactg ctgcgtgcca agacaggcgg cacctatggc gaggacttgg
7801 gtgctgacta caacctctcc caggtgtgtg acgggaaggt gtctgtgcac atcattgagg
7861 gtgaccaccg cacactgctg gagggcagtg gcctggaatc catcatcaac atcatccata
7921 gctccctggc tgagccacga gtgagtgtac gggagggcct cgagcatcat caccaccacc
7981 accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
8041 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
8101 tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt
8161 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
8221 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
8281 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
8341 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
8401 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
8461 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
8521 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
8581 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
8641 tttttctaaa tacattcaaa tatgtattcg ctcatgagac aataaccctg ataaatgctt
8701 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
8761 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
8821 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
8881 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
8941 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
9001 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
9061 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
9121 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
9181 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
9241 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
9301 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
9361 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
9421 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
9481 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
9541 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
9601 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
9661 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
9721 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
9781 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
9841 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
9901 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
9961 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
10021 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
10081 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
10141 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
10201 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
10261 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
10321 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
10381 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
10441 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
10501 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
10561 tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt
10621 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
10681 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
10741 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
10801 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
10861 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
10921 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg
10981 gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg
11041 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt
11101 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac
11161 agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat
11221 ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat
11281 tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg
11341 tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa
11401 cgacaggagc acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt
11461 ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat
11521 tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc
11581 gaaaatgacc cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt
11641 cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
11701 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
11761 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
11821 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
11881 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
11941 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
12001 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac
12061 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
12121 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
12181 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
12241 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
12301 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
12361 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
12421 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
12481 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
12541 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
12601 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
12661 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
12721 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
12781 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
12841 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
12901 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
12961 tcctgcatta ggaagcagcc cag
//