| 质粒名称: | pC001-huLshC2C2-MBP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 79150 |
| 存储实验室: |
Feng Zhang
|
|
载体骨架基本信息 |
|
| 载体名称: | BPV00356 pET21GG2-His(6)-MBP_Flag-Avi |
| 载体出品公司: | Zhang lab |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 6650 bp |
| 完整质粒大小: | 10955 bp |
| 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: | C2c2 |
| 物种来源: | Leptotrichia shahii |
| 插入大小: | 4305 bp |
| 基因又名: | Cas13a |
| 启动子 | T7 |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BsaI(未知破坏) |
| 3' 克隆位点: | BsaI(未知破坏) |
| 5' 测序引物 | atggggaacctgttcggacaca |
|
文献引用方法 |
|
| 材料和方法部分: | pC001 - huLshC2C2-MBP for bacterial expression was a gift from Feng Zhang (爱迪基因 plasmid # 79150) |
| 参考文献部分: | C2c2 is a single-component programmable RNA-guided RNA-targeting CRISPR effector. Abudayyeh OO, Gootenberg JS, Konermann S, Joung J, Slaymaker IM, Cox DB, Shmakov S, Makarova KS, Semenova E, Minakhin L, Severinov K, Regev A, Lander ES, Koonin EV, Zhang F. Science. 2016 Jun 2. pii: aaf5573. 10.1126/science.aaf5573 PubMed 27256883 |
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 pC001 - huLshC2C2-MBP 10951 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Expresses human codon-optimized LshC2c2 for purification in E. coli.
ACCESSION .
VERSION .
KEYWORDS pC001 - huLshC2C2-MBP for bacterial expression
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10951)
AUTHORS Abudayyeh OO, Gootenberg JS, Konermann S, Joung J, Slaymaker IM, Cox
DB, Shmakov S, Makarova KS, Semenova E, Minakhin L, Severinov K,
Regev A, Lander ES, Koonin EV, Zhang F
TITLE C2c2 is a single-component programmable RNA-guided RNA-targeting
CRISPR effector.
JOURNAL Science. 2016 Jun 2. pii: aaf5573.
PUBMED 27256883
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pC001 - huLshC2C2-MBP for bacterial expression|
FEATURES Location/Qualifiers
source 1..10951
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 1012..1031
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(1051..1128)
/gene="lacI"
/label=lacI promoter
primer_bind 1334..1353
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 1437..1456
/label=T7
/note="T7 promoter, forward primer"
promoter 1437..1455
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1456..1480
/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 1495..1517
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 1531..1548
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
regulatory 2503..2512
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2665..2682
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS 2689..2709
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 2710..6876
/codon_start=1
/product="type VI CRISPR effector from Leptotrichia shahii
with RNA-guided RNAse activity (Abudayyeh et al., 2016)"
/label=LshC2c2
/translation="MGNLFGHKRWYEVRDKKDFKIKRKVKVKRNYDGNKYILNINENNN
KEKIDNNKFIRKYINYKKNDNILKEFTRKFHAGNILFKLKGKEGIIRIENNDDFLETEE
VVLYIEAYGKSEKLKALGITKKKIIDEAIRQGITKDDKKIEIKRQENEEEIEIDIRDEY
TNKTLNDCSIILRIIENDELETKKSIYEIFKNINMSLYKIIEKIIENETEKVFENRYYE
EHLREKLLKDDKIDVILTNFMEIREKIKSNLEILGFVKFYLNVGGDKKKSKNKKMLVEK
ILNINVDLTVEDIADFVIKELEFWNITKRIEKVKKVNNEFLEKRRNRTYIKSYVLLDKH
EKFKIERENKKDKIVKFFVENIKNNSIKEKIEKILAEFKIDELIKKLEKELKKGNCDTE
IFGIFKKHYKVNFDSKKFSKKSDEEKELYKIIYRYLKGRIEKILVNEQKVRLKKMEKIE
IEKILNESILSEKILKRVKQYTLEHIMYLGKLRHNDIDMTTVNTDDFSRLHAKEELDLE
LITFFASTNMELNKIFSRENINNDENIDFFGGDREKNYVLDKKILNSKIKIIRDLDFID
NKNNITNNFIRKFTKIGTNERNRILHAISKERDLQGTQDDYNKVINIIQNLKISDEEVS
KALNLDVVFKDKKNIITKINDIKISEENNNDIKYLPSFSKVLPEILNLYRNNPKNEPFD
TIETEKIVLNALIYVNKELYKKLILEDDLEENESKNIFLQELKKTLGNIDEIDENIIEN
YYKNAQISASKGNNKAIKKYQKKVIECYIGYLRKNYEELFDFSDFKMNIQEIKKQIKDI
NDNKTYERITVKTSDKTIVINDDFEYIISIFALLNSNAVINKIRNRFFATSVWLNTSEY
QNIIDILDEIMQLNTLRNECITENWNLNLEEFIQKMKEIEKDFDDFKIQTKKEIFNNYY
EDIKNNILTEFKDDINGCDVLEKKLEKIVIFDDETKFEIDKKSNILQDEQRKLSNINKK
DLKKKVDQYIKDKDQEIKSKILCRIIFNSDFLKKYKKEIDNLIEDMESENENKFQEIYY
PKERKNELYIYKKNLFLNIGNPNFDKIYGLISNDIKMADAKFLFNIDGKNIRKNKISEI
DAILKNLNDKLNGYSKEYKEKYIKKLKENDDFFAKNIQNKNYKSFEKDYNRVSEYKKIR
DLVEFNYLNKIESYLIDINWKLAIQMARFERDMHYIVNGLRELGIIKLSGYNTGISRAY
PKRNGSDGFYTTTAYYKFFDEESYKKFEKICYGFGIDLSENSEINKPENESIRNYISHF
YIVRNPFADYSIAEQIDRVSNLLSYSTRYNNSTYASVFEVFKKDVNLDYDELKKKFKLI
GNNDILERLMKPKKVSVLELESYNSDYIKNLIIELLTKIENTNDTL"
CDS 6877..6924
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 6931..7011
/codon_start=1
/product="three tandem HA epitope tags"
/label=3xHA
/translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
CDS 7035..7058
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 7059..7103
/codon_start=1
/product="peptide tag that allows for enzymatic
biotinylation"
/label=AviTag(TM)
/translation="GLNDIFEAQKIEWHE"
CDS 7119..7136
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(7189..7207)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 7203..7250
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 7287..7742
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7374..7393)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7584..7605
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 7768..7872
/gene="bla"
/label=AmpR promoter
CDS 7873..8733
/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(8091..8110)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8904..9492
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9393..9412
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 9646..9663
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 9678..9817
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(9743..9762)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 9903..9925
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(9919..10110)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(10919..1050)
/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"
ORIGIN
1 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
61 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
121 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
181 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
241 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
301 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
361 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
421 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
481 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
541 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
601 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
661 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
721 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
781 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
841 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
901 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
961 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
1021 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
1081 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
1141 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
1201 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
1261 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
1321 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
1381 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat
1441 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt
1501 taactttaag aaggagatat acatatgggt catcatcatc accatcacaa aatcgaagaa
1561 ggcaaactgg ttatttggat caatggcgat aaaggctata atggtctggc agaagttggc
1621 aaaaaattcg aaaaagatac cggcattaaa gtgaccgttg aacatccgga taaactggaa
1681 gaaaaatttc cgcaggttgc agcaaccggt gatggtccgg atattatctt ttgggcacat
1741 gatcgttttg gtggttatgc acagagcggt ctgctggcag aaattacacc ggataaagca
1801 tttcaggaca aactgtatcc gtttacctgg gatgcagttc gctataacgg taaactgatt
1861 gcatatccga ttgcagttga agcactgagc ctgatctata acaaagatct gctgccgaat
1921 ccgcctaaaa cctgggaaga aattccggca ctggataaag aactgaaagc aaaaggtaaa
1981 agcgcactga tgtttaatct gcaagaaccg tattttacct ggcctctgat tgcagcagat
2041 ggtggctatg cattcaaata tgaaaacggc aaatatgata tcaaagacgt gggtgttgat
2101 aatgcgggtg caaaagccgg tctgaccttt ctggttgatc tgattaaaaa caaacacatg
2161 aacgccgata ccgattatag cattgcagaa gcagcattta acaaaggtga aaccgcaatg
2221 acaattaatg gtccgtgggc atggtcaaat attgatacca gcaaagtgaa ttatggtgtt
2281 accgttctgc cgacatttaa aggtcagccg agcaaaccgt ttgttggtgt tctgagcgca
2341 ggtattaatg cagcaagccc gaacaaagaa ctggcaaaag aatttctgga aaactatctg
2401 ctgaccgatg aaggtctgga agcagtgaat aaagataaac cgctgggtgc agttgcactg
2461 aaaagctatg aagaagaact ggttaaagat ccgcgtattg cagccaccat ggaaaatgca
2521 cagaaaggcg aaattatgcc gaatattccg cagatgagcg cattttggta tgccgttcgt
2581 accgcagtga ttaatgccgc atcaggtcgt cagaccgttg atgaagccct gaaagatgcc
2641 cagaccggtg caagcaccgg atccctggtt ccgcgtggta gcgctagcga aaatctgtat
2701 tttcagggaa tggggaacct gttcggacac aagcggtggt acgaggtgcg agataagaaa
2761 gacttcaaga tcaagcgaaa ggtgaaagtc aagcggaatt atgacgggaa caagtacatt
2821 ctgaacatca acgaaaacaa caacaaagag aagatcgaca acaacaagtt catcagaaag
2881 tacatcaact acaagaaaaa cgataatatt ctgaaggagt tcactaggaa atttcatgca
2941 ggaaatatcc tgttcaaact gaagggcaaa gaagggatca ttagaattga gaataacgac
3001 gacttcctgg agacagagga agtggtgctg tatatcgagg cctacggcaa gagcgagaag
3061 ctgaaagcac tggggatcac aaagaaaaag atcattgacg aggccatcag gcagggcatt
3121 actaaggacg ataaaaagat cgagatcaag cgacaggaga acgaggaaga gatcgaaatt
3181 gacatccggg atgagtacac taataagacc ctgaacgact gctccatcat tctgcgcatc
3241 attgaaaacg atgaactgga gacaaagaag agcatctacg agatcttcaa gaacatcaat
3301 atgagcctgt ataagatcat cgagaagatc atcgaaaacg agacagaaaa ggtgtttgaa
3361 aatagatact acgaagagca cctgagggag aagctgctga aagacgataa gattgacgtg
3421 atcctgacca acttcatgga aatccgggag aagatcaagt ctaatctgga gatcctgggc
3481 ttcgtgaagt tttacctgaa cgtcggcggg gacaaaaaga aaagtaaaaa taagaaaatg
3541 ctggtggaaa agattctgaa catcaatgtg gatctgaccg tcgaggacat tgccgatttc
3601 gtgatcaagg agctggaatt ttggaacatc acaaagcgca ttgagaaagt gaagaaagtc
3661 aataacgagt tcctggagaa gcggagaaat cggacataca tcaagtccta tgtgctgctg
3721 gacaagcacg aaaagtttaa aatcgagaga gaaaacaaga aggataagat cgtgaagttc
3781 tttgtcgaga acattaagaa caactctatc aaggaaaaga ttgagaagat cctggctgag
3841 ttcaagatcg acgagctgat taagaaactg gagaaggaac tgaagaaagg gaactgtgat
3901 accgagatct tcggaatctt taagaagcat tacaaggtga acttcgacag caagaaattt
3961 tccaagaaat ctgatgaaga gaaggagctg tataagatca tctacagata cctgaagggc
4021 agaattgaaa aaatcctggt gaacgagcag aaggtcagac tgaagaaaat ggagaagatc
4081 gagatcgaaa agattctgaa tgaaagtatc ctgtcagaga aaattctgaa gagagtgaaa
4141 cagtatacac tggagcacat tatgtacctg gggaagctga ggcataacga catcgatatg
4201 accacagtga atactgacga tttcagccgc ctgcacgcca aggaagagct ggacctggaa
4261 ctgatcacct tctttgccag cacaaatatg gagctgaaca agatcttttc ccgagaaaac
4321 atcaacaacg acgagaacat cgatttcttt ggaggcgacc gggagaagaa ctatgtgctg
4381 gataagaaaa tcctgaatag taagatcaag atcatccgcg acctggattt catcgataac
4441 aagaacaaca tcacaaacaa cttcattcga aagtttacaa agatcggcac taatgaaagg
4501 aaccgcatcc tgcatgccat ttccaaagag agggacctgc aggggactca ggacgattac
4561 aacaaagtga tcaacatcat tcagaatctg aagatctccg atgaagaggt gagcaaagct
4621 ctgaacctgg acgtggtctt taaggacaag aaaaacatca tcacaaagat caatgacatc
4681 aagatctctg aagagaacaa caacgatatc aagtatctgc ccagcttcag caaagtgctg
4741 cccgaaatcc tgaacctgta ccgcaacaat cccaagaatg agccttttga cacaatcgag
4801 actgaaaaaa ttgtgctgaa cgctctgatc tacgtcaata aggagctgta taagaaactg
4861 atcctggagg acgatctgga agagaacgag tccaagaata tcttcctgca ggaactgaag
4921 aaaaccctgg gcaacattga cgaaatcgat gagaacatca tcgagaacta ctacaagaac
4981 gcacagattt ctgccagtaa ggggaacaac aaggcaatca agaaatatca gaagaaagtg
5041 atcgagtgct acattggata tctgcgcaaa aactacgaag agctgttcga cttttcagac
5101 ttcaagatga acatccagga aatcaagaaa cagattaagg acatcaacga taacaagact
5161 tatgagcgga tcaccgtgaa aaccagcgac aaaaccattg tcatcaacga cgatttcgag
5221 tacatcattt ctatctttgc actgctgaac agtaatgccg tgattaataa gatccgaaac
5281 agattcttcg ccaccagcgt gtggctgaac acctcagaat accagaatat cattgacatc
5341 ctggatgaga ttatgcagct gaataccctg cggaacgaat gcatcacaga gaactggaat
5401 ctgaacctgg aagagttcat tcagaagatg aaagagatcg aaaaggattt cgacgacttc
5461 aagatccaga ctaagaaaga aatcttcaac aactactacg aggacatcaa gaacaacatt
5521 ctgaccgagt ttaaagacga tatcaacggc tgtgatgtgc tggaaaagaa actggagaag
5581 attgtcatct tcgacgatga aaccaagttc gagatcgaca agaaatccaa catcctgcag
5641 gatgaacaga gaaagctgtc taacatcaac aagaaggacc tgaagaaaaa ggtggatcag
5701 tatatcaagg acaaagatca ggagatcaag tctaaaatcc tgtgcaggat cattttcaac
5761 agtgactttc tgaaaaagta caagaaggaa atcgacaatc tgattgagga tatggagtct
5821 gaaaatgaga acaagttcca ggagatctac tatcccaagg aacggaagaa cgagctgtat
5881 atctacaaaa agaatctgtt cctgaacatc ggaaatccta actttgacaa gatctacggc
5941 ctgattagca acgacatcaa gatggccgat gctaaattcc tgtttaatat cgatggaaag
6001 aacatcagaa agaacaaaat cagtgagatc gacgctattc tgaagaatct gaacgataaa
6061 ctgaacggct actcaaagga atacaaggag aagtacatca aaaagctgaa ggagaacgac
6121 gatttctttg caaagaacat ccagaataag aactacaaat ccttcgaaaa ggactataac
6181 cgcgtgtctg agtacaaaaa gattcgagat ctggtcgagt tcaactatct gaacaaaatc
6241 gagtcctacc tgattgacat caactggaag ctggctattc agatggcaag attcgaaagg
6301 gatatgcact atatcgtgaa tggactgagg gagctgggca tcattaagct gtcaggctat
6361 aacaccggga tcagcagggc atacccaaag cgcaatggaa gcgacggctt ttacactacc
6421 acagcctact acaagttctt tgatgaagag tcctacaaga agttcgagaa gatttgctac
6481 gggtttggaa tcgacctgag cgaaaattcc gagatcaaca agcctgaaaa tgagagcatt
6541 cggaactata tctcccattt ctacatcgtg agaaatccat ttgccgacta cagtattgct
6601 gagcagatcg atcgggtgag caacctgctg tcatatagca cacgctacaa caattcaact
6661 tatgccagcg tgttcgaagt ctttaaaaag gacgtgaatc tggactacga tgagctgaaa
6721 aagaaattca aactgatcgg caacaatgat attctggagc gcctgatgaa gcccaagaaa
6781 gtgagcgtcc tggaactgga gtcctacaac agtgactaca ttaagaatct gatcattgaa
6841 ctgctgacca aaatcgagaa tactaacgat accctgaaaa ggccggcggc cacgaaaaag
6901 gccggccagg caaaaaagaa aaagggatcc tacccatacg atgttccaga ttacgcttat
6961 ccctacgacg tgcctgatta tgcataccca tatgatgtcc ccgactatgc ctaagcctcg
7021 agggtggtgg tagcgattat aaagatgatg atgataaagg cctgaacgat atctttgaag
7081 cccagaaaat tgaatggcac gagtaaaagc ttgtcgagca ccaccaccac caccactgag
7141 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
7201 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
7261 gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
7321 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
7381 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
7441 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
7501 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
7561 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
7621 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
7681 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
7741 ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
7801 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
7861 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
7921 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
7981 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
8041 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
8101 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
8161 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
8221 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
8281 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
8341 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
8401 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
8461 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
8521 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
8581 gagcgtggta gtcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
8641 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
8701 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
8761 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
8821 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
8881 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
8941 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
9001 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
9061 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
9121 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
9181 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
9241 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
9301 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
9361 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
9421 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
9481 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
9541 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
9601 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
9661 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
9721 caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
9781 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
9841 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
9901 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct
9961 gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc
10021 gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat
10081 gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt
10141 catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga
10201 tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg
10261 ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt
10321 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
10381 tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc
10441 tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc
10501 attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac
10561 gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
10621 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
10681 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
10741 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
10801 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
10861 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
10921 actgcccgct ttccagtcgg gaaacctgtc g
//
