| 质粒名称: | D. vulgaris Cascade/I-C (Cas5c-Cas8c-Cas7)/pHMGWA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 81185 |
| 存储实验室: | Jennifer Doudna |
|
载体骨架基本信息 |
|
| 载体名称: | pHMGWA |
| 载体出品公司: | ThermoFisher |
| 空载体大小: | 8236 bp |
| 完整质粒大小: | 10041 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达，CRISPR |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称1: | Cas5c |
| 物种来源1: | 希尔登堡普通脱硫弧菌街 |
| 插入大小1: | 680 bp |
| GenBank编号1 | 2781521 YP_009170 |
| 启动子1: | T7 |
| 标签/融合蛋白1: | 6xHis-MBP-TEV蛋白酶位点（插入物上的N末端） |
| 克隆方法1: | Gateway Cloning |
| 5’测序引物1 | MBP forward |
| 3’测序引物1 | T7 terminator |
| 插入基因名称2: | Cas8c |
| 物种来源2: | 希尔登堡普通脱硫弧菌街 |
| 插入大小2: | 1839 bp |
| GenBank编号2 | 2781582 YP_009171.1 |
| 启动子2: | T7 |
| 标签/融合蛋白2: | -- |
| 克隆方法2: | Gateway Cloning |
| 5’测序引物2 | MBP forward |
| 3’测序引物2 | T7 terminator |
| 插入基因名称3: | Cas7 |
| 物种来源3: | 希尔登堡普通脱硫弧菌街 |
| 插入大小3: | 873 bp |
| GenBank编号3 | 2781595 YP_009172.1 |
| 启动子3: | T7 |
| 克隆方法3: | Gateway Cloning |
| 5’测序引物3 | MBP forward |
| 3’测序引物3 | T7 terminator |
|
文献引用方法 |
|
| 材料和方法部分: | D. vulgaris Cascade/I-C (Cas5c-Cas8c-Cas7)/pHMGWA was a gift from Jennifer Doudna (Addgene plasmid # 81185 ; http://n2t.net/addgene:81185 ; RRID:Addgene_81185) |
| 参考文献部分: | DNA Targeting by a Minimal CRISPR RNA-Guided Cascade. Hochstrasser ML, Taylor DW, Kornfeld JE, Nogales E, Doudna JA. Mol Cell. 2016 Sep 1;63(5):840-51. doi: 10.1016/j.molcel.2016.07.027. 10.1016/j.molcel.2016.07.027 PubMed 27588603 |
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
 pHMGWA质粒图谱.png)
载体简介
载体序列
LOCUS D. vulgaris Cascade/I-C (Cas5c-Cas8c-Cas7)/pHMGWA 10037 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses Desulfovibrio vulgaris Cascade/I-C complex subunits in E.
coli with TEV-cleavable N-terminal His-MBP tag on Cas5c.
ACCESSION .
VERSION .
KEYWORDS D. vulgaris Cascade/I-C (Cas5c-Cas8c-Cas7)/pHMGWA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10037)
AUTHORS Hochstrasser ML, Taylor DW, Kornfeld JE, Nogales E, Doudna JA
TITLE DNA Targeting by a Minimal CRISPR RNA-Guided Cascade.
JOURNAL Mol Cell. 2016 Sep 1;63(5):840-51. doi:
10.1016/j.molcel.2016.07.027.
PUBMED 27588603
REFERENCE 2 (bases 1 to 10037)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..10037
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 27..46
/label=T7
/note="T7 promoter, forward primer"
promoter 27..45
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 46..70
/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 85..107
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 126..143
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 150..1247
/codon_start=1
/gene="malE (mutated)"
/product="maltose binding protein from E. coli"
/label=MBP
/note="This version of the gene does not encode a signal
sequence, so MBP will remain in the cytosol."
/translation="KTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
YAVRTAVINAASGRQTVDEALKDAQT"
regulatory 1104..1113
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind 1221..1244
/label=MBP-F
/note="Maltose binding protein, forward primer"
protein_bind 1266..1290
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 1311..1331
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
protein_bind complement(4751..4775)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 4789..4806
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(4859..4877)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 4873..4920
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 4957..5412
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5044..5063)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5254..5275
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 5438..5542
/gene="bla"
/label=AmpR promoter
CDS 5543..6403
/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(5761..5780)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6574..7162
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7063..7082
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7316..7333
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 7348..7487
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(7413..7432)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7573..7595
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(7589..7780)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(8589..9671)
/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 9633..9652
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(9672..9749)
/gene="lacI"
/label=lacI promoter
primer_bind 9955..9974
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 agatccagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
61 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accatgggca
121 gcagccatca tcatcatcat cacggtacca aaactgaaga aggtaaactg gtaatctgga
181 ttaacggcga taaaggctat aacggtctcg ctgaagtcgg taagaaattc gagaaagata
241 ccggaattaa agtcaccgtt gagcatccgg ataaactgga agagaaattc ccacaggttg
301 cggcaactgg cgatggccct gacattatct tctgggcaca cgaccgcttt ggtggctacg
361 ctcaatctgg cctgttggct gaaatcaccc cggacaaagc gttccaggac aagctgtatc
421 cgtttacctg ggatgccgta cgttacaacg gcaagctgat tgcttacccg atcgctgttg
481 aagcgttatc gctgatttat aacaaagatc tgctgccgaa cccgccaaaa acctgggaag
541 agatcccggc gctggataaa gaactgaaag cgaaaggtaa gagcgcgctg atgttcaacc
601 tgcaagaacc gtacttcacc tggccgctga ttgctgctga cgggggttat gcgttcaagt
661 atgaaaacgg caagtacgac attaaagacg tgggcgtgga taacgctggc gcgaaagcgg
721 gtctgacctt cctggttgac ctgattaaaa acaaacacat gaatgcagac accgattact
781 ccatcgcaga agctgccttt aataaaggcg aaacagcgat gaccatcaac ggcccgtggg
841 catggtccaa catcgacacc agcaaagtga attatggtgt aacggtactg ccgaccttca
901 agggtcaacc atccaaaccg ttcgttggcg tgctgagcgc aggtattaac gccgccagtc
961 cgaacaaaga gctggcaaaa gagttcctcg aaaactatct gctgactgat gaaggtctgg
1021 aagcggttaa taaagacaaa ccgctgggtg ccgtagcgct gaagtcttac gaggaagagt
1081 tggcgaaaga tccacgtatt gccgccacca tggaaaacgc ccagaaaggt gaaatcatgc
1141 cgaacatccc gcagatgtcc gctttctggt atgccgtgcg tactgcggtg atcaacgccg
1201 ccagcggtcg tcagactgtc gatgaagccc tgaaagacgc gcagactggt accggatctt
1261 acatcacaag tttgtacaaa aaagcaggct ccgcggccgc cgtactcgag gaaaacctgt
1321 attttcaggg ctccttcacc atgacacatg gggctgttaa gacatacggc atcagactgc
1381 gggtatgggg cgactacgcc tgcttcacgc ggccggagat gaaggtggaa cgggtgtcat
1441 acgatgtcat gcccccttcc gccgcacggg gcatccttga ggccatccac tggaagccgg
1501 ccatcaggtg gattgtggac aggattcatg tgctgcggcc catcgtcttc gacaacgtgc
1561 ggcgcaacga ggtcagttcg aagataccca agcccaatcc ggcaacggcc atgcgcgaca
1621 ggaagccgct ctactttctg gtcgatgacg gcagcaaccg ccagcaacgg gcggcgaccc
1681 tgctgcgcaa cgtcgactat gtcatcgaag cccacttcga actgacagac aaggccggag
1741 cggaagacaa cgcgggcaag catctggaca tcttcaggcg tcgtgcgcgc gccgggcagt
1801 cgttccagca gccatgcctc ggctgccgcg agtttccggc gtcgttcgaa ctgctggagg
1861 gggatgtgcc gctctcgtgc tacgcgggcg agaagcgcga ccttggctac atgctgctcg
1921 acatcgactt cgaacgcgac atgacgccgc tgttcttcaa ggccgtcatg gaagacggag
1981 tgataacgcc cccgtcgcgc acgtcaccgg aggtgagggc atgatcctgc aggcattgca
2041 tggctattac cagcgcatga gcgcagaccc cgatgcgggg atgccgccct acgggaccag
2101 catggagaac atctcgttcg cgctggtgct ggatgcgaaa gggacgctac ggggcatcga
2161 ggacttgcgg gaacaggaag gcaagaagct gcggccccgc aagatgctgg ttcctatcgc
2221 ggaaaagaag gggaacggca tcaaaccgaa cttcctgtgg gagaacacca gttatattct
2281 gggggtggac gccaagggca agcaggaacg tacagacaag tgccacgccg cgttcattgc
2341 ccacatcaag gcctactgtg acactgcaga ccaagacctt gccgccgttc tgcagttcct
2401 tgaacacggc gaaaaagacc tctccgcttt tccggtttcg gaagaggtca tcggctccaa
2461 tatcgtcttt cgcatcgagg gcgaaccggg cttcgtccat gagcgtccgg ctgcgcggca
2521 ggcttgggcc aactgcctga accggcggga gcaagggcta tgtggtcaat gcctcatcac
2581 cggggagcgc cagaagccga ttgcccaact ccatccgtcg atcaaaggcg ggcgagacgg
2641 cgttcgcggt gcacaggctg ttgcctcgat agtgtccttt aacaacaccg cctttgaatc
2701 ttacgggaaa gaacagtcga tcaatgcacc cgtcagtcaa gaagcggcat tctcttatgt
2761 cacagccctg aattacctgc tcaacccgtc aaatcgacag aaggtcacca tcgctgacgc
2821 caccgtggtc ttctgggccg aacgcagcag cccggcagag gacatctttg caggcatgtt
2881 cgacccgcct tccaccacgg cgaaaccgga gtcctcgaac gggacgccgc cggaagacag
2941 tgaagaaggc agccagcctg acacagccag agacgacccc catgccgctg ccagaatgca
3001 cgacctgctt gtggccatcc gttccggcaa gcgcgccacg gacatcatgc ccgacatgga
3061 cgagtccgtg cgcttccatg tgcttggcct gtcgcccaac gccgcccgcc tgtccgtccg
3121 cttctgggag gtcgataccg tcggtcacat gctggacaag gtggggcggc actaccgtga
3181 actcgagatc attccgcaat tcaacaacga gcaggagttc ccgtctcttt caaccctgct
3241 gcggcagact gccgtcttga acaagaccga gaacatctcg cctgtacttg caggcgggct
3301 attcagggcc atgctcaccg gcgggcccta tccgcaaagc ctgttgcccg ccgtgcttgg
3361 acgcatccgc gccgagcacg ccagaccaga agataaaagc cgctataggc ttgaagttgt
3421 gacgtattat cgagccgcct tgatcaaggc gtatctcata agaaacagaa agctggaggt
3481 gcccgtgtct ctcgaccccg caagaacaga ccgcccgtac ctgctcggac gactcttcgc
3541 cgttctggaa aaagcgcagg aagatgccgt gcccggtgcg aacgccacca tcaaggaccg
3601 ctatctggcc tccgcatcgg ccaatccggg gcaggtattc cacatgctgc tcaagaatgc
3661 gtccaaccac acggccaaac tccgcaagga ccctgaaaga aaggggtcgg ccatccacta
3721 cgaaatcatg atgcaagaga ttatcgacaa catcagcgat tttccggtga ccatgtcgtc
3781 cgacgaacaa ggcctgttca tgatcggcta ctaccaccag cgcaaggccc ttttcaccaa
3841 gaagaacaag gagaactagc catgaccgcc attgccaaca gatacgagtt cgtgctgctt
3901 ttcgatgtgg agaacggcaa ccccaacggc gaccccgacg cgggcaacat gccgcgcatc
3961 gaccccgaga ccgggcatgg tctggtcacc gacgtgtgcc tgaagcgcaa gatacgcaac
4021 catgtggccc tgacaaaaga aggtgccgag cggttcaata tctatattca ggaaaaggcc
4081 attctcaacg aaacccacga acgcgcctac acggcctgcg acctgaaacc cgaacccaag
4141 aaactgccca agaaggtcga agacgccaag agggtcaccg actggatgtg cacgaacttc
4201 tacgacatcc gcacgttcgg cgccgtcatg accacagagg tcaactgcgg gcaggtgcgc
4261 ggccccgtgc agatggcctt tgccagaagt gtggaacccg tcgtgccgca agaggtgagc
4321 atcacccgca tggcagtcac cacgaaggca gaggccgaaa agcagcaggg agacaaccgc
4381 accatgggtc gcaagcacat cgtgccctac ggcctgtatg tggcgcacgg cttcatctcc
4441 gcaccgctgg cagagaagac aggcttctcc gatgaagacc tgacgctgtt ctgggacgcc
4501 ctcgtcaaca tgttcgagca cgaccgttcg gcggcacgcg gcctcatgag cagccgcaaa
4561 ctcatcgtct tcaagcacca gaacaggctc ggcaacgccc ccgcccacaa gttgttcgac
4621 ctcgtgaagg tcagccgtgc agagggttcg tccggccccg cccggtcgtt tgccgactat
4681 gcggtgacag tggggcaggc ccccgaaggc gtagaggtca aagagatgct gtagaagggt
4741 gggcgcgccg acccagcttt cttgtacaaa gtggtgatgt acctcgagca ccaccaccac
4801 caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
4861 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
4921 ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa
4981 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
5041 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
5101 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
5161 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
5221 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
5281 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
5341 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
5401 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
5461 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
5521 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
5581 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
5641 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
5701 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
5761 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
5821 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
5881 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
5941 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
6001 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
6061 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
6121 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
6181 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
6241 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
6301 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
6361 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
6421 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
6481 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
6541 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
6601 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
6661 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
6721 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
6781 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
6841 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
6901 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
6961 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
7021 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
7081 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
7141 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
7201 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
7261 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
7321 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
7381 cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
7441 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
7501 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
7561 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
7621 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
7681 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
7741 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
7801 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
7861 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
7921 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
7981 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg
8041 tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc
8101 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta
8161 tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg
8221 acaggagcac gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg gcctgcttct
8281 cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc
8341 cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga
8401 aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca
8461 taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg
8521 ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg
8581 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
8641 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc
8701 accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc
8761 aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc
8821 gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca
8881 acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca
8941 accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg
9001 gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga
9061 tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac
9121 agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct
9181 tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc
9241 ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta
9301 atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg
9361 acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat
9421 ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca
9481 atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc
9541 tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc
9601 accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt
9661 actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg
9721 cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc
9781 ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa
9841 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
9901 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
9961 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
10021 tccggcgtag aggatcg
//
