| 质粒名称: | pX330S-2-PITCh |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 63670 |
| 存储实验室: | Takashi Yamamoto |
|
载体骨架基本信息 |
|
| 载体名称: | pUC ori vector |
| 载体出品公司: | -- |
| 载体抗性: | 大观霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | humanized S. pyogenes Cas9 |
| 物种来源: | 化脓链球菌 |
| 插入大小: | 4272 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CBh |
| 标签/融合蛋白: | 3xFLAG (N terminal on insert) |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | T7 (TAATACGACTCACTATAGGG) |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pX330S-2-PITCh was a gift from Takashi Yamamoto (Addgene plasmid # 63670 ; http://n2t.net/addgene:63670 ; RRID:Addgene_63670) |
| 参考文献部分: | MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the PITCh systems. Sakuma T, Nakade S, Sakane Y, Suzuki KT, Yamamoto T. Nat Protoc. 2016 Jan;11(1):118-33. doi: 10.1038/nprot.2015.140. Epub 2015 Dec 17. 10.1038/nprot.2015.140 PubMed 26678082 |
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 pX330S-2-PITCh 8700 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses Cas9 nuclease and the PITCh-gRNA.
ACCESSION .
VERSION .
KEYWORDS pX330S-2-PITCh
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8700)
AUTHORS Sakuma T, Nakade S, Sakane Y, Suzuki KT, Yamamoto T
TITLE MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the
PITCh systems.
JOURNAL Nat Protoc. 2016 Jan;11(1):118-33. doi: 10.1038/nprot.2015.140. Epub
2015 Dec 17.
PUBMED 26678082
REFERENCE 2 (bases 1 to 8700)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8700
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 8..293
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
promoter 295..572
/label=chicken beta-actin promoter
intron 573..801
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
regulatory 813..822
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 822..887
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 894..914
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 939..5039
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 5040..5087
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
primer_bind complement(5115..5132)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5121..5328
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 5337..5477
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 5337..5466
/label=AAV2 ITR
rep_origin 5552..6007
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5639..5658)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5849..5870
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6024..6043)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6143..6165
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6203..6221)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
CDS 6628..7419
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 7602..8190
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8091..8110
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 8268..8508
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 8268..8288
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 8439..8458
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 8537..8612
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
ORIGIN
1 tgagacccgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
61 gcccattgac gtcaatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
121 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
181 ctattgacgt caatgacggt aaatggcccg cctggcattg tgcccagtac atgaccttat
241 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg
301 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt
361 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggggcgcg
421 cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg
481 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc
541 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgacgctg ccttcgcccc
601 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
661 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagct gagcaagagg
721 taagggttta agggatggtt ggttggtggg gtattaatgt ttaattacct ggagcacctg
781 cctgaaatca ctttttttca ggttggaccg gtgccaccat ggactataag gaccacgacg
841 gagactacaa ggatcatgat attgattaca aagacgatga cgataagatg gccccaaaga
901 agaagcggaa ggtcggtatc cacggagtcc cagcagccga caagaagtac agcatcggcc
961 tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
1021 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
1081 gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca
1141 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
1201 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
1261 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
1321 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
1381 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
1441 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
1501 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
1561 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc
1621 tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga
1681 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg
1741 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
1801 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
1861 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc
1921 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
1981 agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc
2041 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac
2101 tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca
2161 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt
2221 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc
2281 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga
2341 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc
2401 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc
2461 ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat
2521 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg
2581 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
2641 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
2701 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
2761 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
2821 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
2881 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
2941 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
3001 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
3061 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
3121 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
3181 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
3241 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
3301 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
3361 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
3421 acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg
3481 acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca
3541 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg
3601 ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga
3661 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
3721 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc
3781 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
3841 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
3901 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg
3961 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
4021 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
4081 agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa
4141 ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca
4201 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt
4261 ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta
4321 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag
4381 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca
4441 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag
4501 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg
4561 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc
4621 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc
4681 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga
4741 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca
4801 aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata
4861 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca
4921 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc
4981 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca
5041 aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa gaaaaagtaa gaattcctag
5101 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
5161 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
5221 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
5281 ggacagcaag ggggaggatt gggaagagaa tagcaggcat gctggggagc ggccgcagga
5341 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
5401 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
5461 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
5521 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
5581 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
5641 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
5701 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
5761 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
5821 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
5881 aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg
5941 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
6001 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
6061 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
6121 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
6181 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
6241 ataataatgg tttcttagac gtcaggtggc acttccagcc aggacagaaa tgcctcgact
6301 tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc
6361 cagtggacat aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc
6421 aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg
6481 gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa gcagcaagcg
6541 cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac gcagcagggc
6601 agtcgcccta aaacaaagtt aaacattatg agggaagcgg tgatcgccga agtatcgact
6661 caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt gctggccgta
6721 catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat tgatttgctg
6781 gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg
6841 gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt caccattgtt
6901 gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca atttggagaa
6961 tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga cattgatctg
7021 gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc agcggcggag
7081 gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga aaccttaacg
7141 ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc
7201 cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg
7261 gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag acaggcttat
7321 cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga atttgtccac
7381 tacgtgaaag gcgagatcac caaggtagtc ggcaaataaa agcattggta actgtcagac
7441 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
7501 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
7561 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
7621 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
7681 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
7741 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
7801 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
7861 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
7921 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
7981 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
8041 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
8101 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
8161 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
8221 ctggcctttt gctggccttt tgctcaggtc tcactctaga ggcatgtgag ggcctatttc
8281 ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata attggaatta
8341 atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc
8401 ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa
8461 cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccgcat
8521 cgtacgcgta cgtgttgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
8581 atcaacttga aaaagtggca ccgagtcggt gcttttttgt tttagagcta gaaatagcaa
8641 gttaaaataa ggctagtccg tttttagcgc gtgcgccaat tctgcagaca aatggctcta
//
