| 质粒名称: | pX601-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA;U6::BsaI-sgRNA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 61591 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV |
| 载体出品公司: | Clontech |
| 空载体大小: | 7446bp |
| 完整质粒大小: | 7446bp |
| 载体类型: | 哺乳细胞表达, AAV, CRISPR |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | Stbl3 |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称1: | hSaCas9/hSauCas9 |
| 物种来源: | Staphylococcus aureus |
| 插入大小: | 3345 bp |
| GenBank编号 | CCK74173.1 |
| 标签/融合蛋白: |
NLS (N terminal on insert)
NLS (C terminal on insert)
3xHA (C terminal on insert)
|
| 插入基因名称2: | Chimeric guide for SaCas9 |
| 物种来源： | Staphylococcus aureus |
| 插入大小： | 101bp |
|
文献引用方法 |
|
| 材料和方法部分: | pX601-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA;U6::BsaI-sgRNA was a gift from Feng Zhang (Addgene plasmid # 61591 ; http://n2t.net/addgene:61591 ; RRID:Addgene_61591) |
| 参考文献部分: | In vivo genome editing using Staphylococcus aureus Cas9. Ran FA, Cong L, Yan WX, Scott DA, Gootenberg JS, Kriz AJ, Zetsche B, Shalem O, Wu X, Makarova KS, Koonin EV, Sharp PA, Zhang F. Nature. 2015 Apr 1. doi: 10.1038/nature14299. 10.1038/nature14299 PubMed 25830891 |
pX601-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA;U6::BsaI-sgRNA
买家导航
pX601-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA;U6::BsaI-sgRNA载体质粒基本信息
pX601-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA;U6::BsaI-sgRNA质粒图谱载体图谱和pX601-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA;U6::BsaI-sgRNA载体序列质粒序列多克隆位点信息
pX601-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA;U6::BsaI-sgRNA质粒载体简介
pX601-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA;U6::BsaI-sgRNA质粒序列载体序列
LOCUS pX601-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA;U6::BsaI-sgRNA 7446 bp DNA circular SYN 14-APR-2025
DEFINITION A single vector AAV-Cas9 system containing Cas9 from Staphylococcus
aureus (SaCas9) and its sgRNA..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7446)
AUTHORS Ran FA, Cong L, Yan WX, Scott DA, Gootenberg JS, Kriz AJ, Zetsche B,
Shalem O, Wu X, Makarova KS, Koonin EV, Sharp PA, Zhang F
TITLE In vivo genome editing using Staphylococcus aureus Cas9.
JOURNAL Nature. 2015 Apr 1. doi: 10.1038/nature14299.
PUBMED 25830891
REFERENCE 2 (bases 1 to 7446)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature.
2015 Apr 1. doi: 10.1038/nature14299."
COMMENT Sequence Label:
pX601-AAV-CMV::NLS-SaCas9-NLS-3xHA-bGHpA;U6::BsaI-sgRNA
FEATURES Location/Qualifiers
source 1..7446
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(869..889)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
regulatory 892..901
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter complement(920..1123)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(949..969)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 1124..1503
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
repeat_region 1527..1656
/label=AAV2 ITR (alternate)
/note="Functional equivalent of wild-type AAV2 ITR"
rep_origin complement(1718..2306)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1798..1817)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(2477..3268,3269..3337))
/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 3100..3119
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3338..3442)
/gene="bla"
/label=AmpR promoter
primer_bind 3510..3528
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(3566..3588)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 3688..3707
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
rep_origin complement(3724..4179)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3861..3882)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4073..4092
/label=F1ori-R
/note="F1 origin, reverse primer"
repeat_region 4254..4394
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 4265..4394
/label=AAV2 ITR
misc_RNA complement(4408..4483)
/label=Sa gRNA scaffold
/note="guide RNA scaffold for the Staphylococcus aureus
CRISPR/Cas9 system"
promoter complement(4504..4752)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA
(Domitrovich & Kunkel, 2003)"
primer_bind complement(4562..4581)
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
primer_bind complement(4732..4752)
/label=hU6-F
/note="Human U6 promoter, forward primer"
polyA_signal 4759..4966
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 4955..4972
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
CDS complement(5000..5080)
/codon_start=1
/product="three tandem HA epitope tags"
/label=3xHA
/translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
CDS complement(5087..5134)
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS complement(join(5135..7446,1..844))
/codon_start=1
/product="Cas9 endonuclease from the Staphylococcus aureus
Type II CRISPR/Cas system"
/label=SaCas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="KRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEG
RRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEF
SAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEV
RGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGW
KDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQ
IIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEI
IENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAI
NLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIK
VINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAK
YLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQ
EENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFS
VQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERN
KGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIF
ITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKDN
DKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKD
NGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNL
DVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLLN
RIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQI
IKKG"
ORIGIN
1 tctggaactt ctcgtaatat tccagcttct cgttctcgtc cctggtgatc acgagattgt
61 tcaggtcgtt cagggcgttg tacaggtcgg cgttgtaggc gtacttcacg ctccgcagtt
121 cctcggggaa gtaggtgcag tggcccatca gcatctcgta ccattctttg atgtccttcc
181 agccgaaggg gctgccctcg ccaggtccct catagtaggt ccgccgggtt tccagcaggt
241 cgatgtaggt gtcgatgaag ctctggtcca gctggtggta ggccttctgc accttcagca
301 gctgtttggc ttctttcacg tagtcgctgg tcttgaatct gttgatgctg ccccgcactt
361 cgccgtcttt cttcagccgt tccagctgca gttcggccac gtatttctct tccagggcct
421 tgctgttccg gctgatctgc tctttggtgg acagctcgtt gccggtgtcc tcttccacct
481 cgttcacgtt gtgcacgcct cttctcttgg ccaggtgcag cagggcggca gagaactctt
541 cctcgctcag cttctggctc aggcccttca ctctggcctc gtaggggttg atgccgctca
601 gctcgctgtg gtcggtcagc aggttgtagt cgaacagcag cttcttcact ctctggattc
661 tatgccgcct ccgccgcttc agccttctgg cgcctctctt gctccgcctg ccctcgttgt
721 tttccacgtt ggcctctttg aacagccgca cgccggcatc gatcacgtcc cgtgtctcgt
781 agtcgatgat gccgtagccc acgctggtga tgccgatgtc caggcccagg atgtagttcc
841 gcttggctgc tgggactccg tggataccga ccttccgctt cttctttggg gccatggtgg
901 caccggtagt tagccagaga gctctgctta tatagacctc ccaccgtaca cgcctaccgc
961 ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt
1021 gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa
1081 accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga
1141 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
1201 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca
1261 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
1321 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt
1381 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat
1441 atgggctatg aactaatgac cccgtaattg attactatta ataactagtc aataatcaat
1501 gtcaacgcct cgagtctaga ggccgcagga acccctagtg atggagttgg ccactccctc
1561 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg
1621 cctcagtgag cgagcgagcg cgcagctgcc tgcaggacat gtgagcaaaa ggccagcaaa
1681 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
1741 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
1801 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
1861 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
1921 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
1981 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
2041 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
2101 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
2161 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
2221 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
2281 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
2341 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
2401 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
2461 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
2521 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
2581 gcttaccatc tggccccagt gctgcaatga taccgcggct tccacgctca ccggctccag
2641 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
2701 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
2761 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
2821 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
2881 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
2941 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
3001 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
3061 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
3121 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
3181 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
3241 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
3301 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
3361 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
3421 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
3481 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
3541 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
3601 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
3661 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
3721 ataaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
3781 cattttttaa ccaatagacc gaaatcggca aaatccctta taaatcaaaa gaatagcccg
3841 agatagagtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
3901 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
3961 ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
4021 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
4081 aagcgaaagg agcgggcgct aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
4141 ccacacccgc cgcgcttaat gcgccgctac agggcgcgta ctatggttgc tttgacgtat
4201 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gcccctgcag
4261 gcagctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac
4321 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
4381 cactaggggt tcctgcggcc gcaaaaatct cgccaacaag ttgacgagat aaacacggca
4441 ttttgccttg ttttagtaga ttctgtttcc agagtactaa aactgagacc tgccgtggtc
4501 tccggtgttt cgtcctttcc acaagatata taaagccaag aaatcgaaat actttcaagt
4561 tacggtaagc atatgatagt ccattttaaa acataatttt aaaactgcaa actacccaag
4621 aaattattac tttctacgtc acgtattttg tactaatatc tttgtgttta cagtcaaatt
4681 aattccaatt atctctctaa cagccttgta tcgtatatgc aaatatgaag gaatcatggg
4741 aaataggccc tcggtacctc cccagcatgc ctgctattct cttcccaatc ctcccccttg
4801 ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca atgcgatgca
4861 atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt caaggaaggc
4921 acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga ggctgatcag
4981 cgagctctag gaattcttaa gcgtaatctg gaacatcgta tgggtaagcg taatctggaa
5041 catcgtatgg gtaagcgtaa tctggaacat cgtatgggta ggatcccttt ttcttttttg
5101 cctggccggc ctttttcgtg gccgccggcc ttttgccctt tttgatgatc tgagggtgct
5161 tcttagattt cacttcatac aggttgccca gaatgtctgt gctgtacttc ttaatgctct
5221 gggtcttgga ggcgattgtc ttaatgatcc tggggggcct cttgtcgttc atgttttcca
5281 ggtactcgcg gtaggtgatg tcgatcatgt tcacttcgat ccggttcagc aggtcgttgt
5341 tcacgccgat cactctatac agctcgccgt tgatcttgat cagatcgttg ttgtagaagg
5401 aggcgataaa ctcggcctgg ttgctgatct tcttcagctt cttagcttcc tcatagcact
5461 tgctattcac ttcgtagtag ttttcttttt tgatcacatc cagattcttc acggtcacga
5521 acttgtacac gccattgtcc aggtacacgt cgaatctgta gggcttcagg gacagcttca
5581 cgaccttgtt tctgctgttg gggtagtcgt cggtgatgtc cagatgggcg ttcagtttgt
5641 tgccgtaata cttaatcttc ttgatcacgg ggccgttgtc ctttttggag tacttggtca
5701 ggtagttccc ggtttcctcg tagtacttgt acaggggatt cttctcgtcg ccgtactgtt
5761 ccataatcag cttcagtttc tggtaggtct gggggtcgtg gtggtacatc agcagctttt
5821 cggggctctt gttgatcagc tttttcagct tgtcattgtc cttgtcgtac aggccgttca
5881 gattgttcac gatcagggtg ttgcccttgt cgtccttccg ggtggagtac agggtgtcgt
5941 taatcagctc tctattaggc ttcttgtcca cccggtggct gtacttgtag tccttgaagt
6001 ccttaatgtg cttgatctgg tggggggtga tgaagatctc tttgtactcc tgctcggttt
6061 cgatctcggg catgctctcg gcctgctttt cctcgaacat ctggttttcc atcacttttt
6121 tggccttgtc cagtttcttc cactctttga agatgaaatc ggcgttggca atgatcaggg
6181 cgtcctcggc gtggtgcttg taccccttgt tccgctcttt cttaaacttc cacttccgcc
6241 gcagaaagct ggtgaagccg ccattgatgg acttcacttt cacgtccagg ttgttcactc
6301 tgaagtagct ccgcagcagg ttcatcaggc ctctggtggc gtatctggta tccaccaggt
6361 tccggttgat gaagtctttc tgcacggaga acctgttgat gtcccgttct tccagcagat
6421 actctttctt ggtcttgctg attctgccct tgcccttggc cagattcagg atgtgcttct
6481 tgaaggtttc gtagctgatc ttgctgtcgc tgctgctcag gtactggaat ggggtccggt
6541 tgcccttctt gctgttttct tcctgcttca cgagcacctt gttgttgaag ctgttgtcga
6601 aggacacgct tctggggatg atgtggtcca cctcatagtt gaaggggttg ttcagcagat
6661 cttccagagg gatggcttcc aggctgtaca ggcacttgcc ttcctgcatg tcgtgcagct
6721 tgatcttctc gatcaggtac ttggcgttct ctttgccggt ggtccggatg atttcctcga
6781 tccgctcgtt ggtctgccgg ttccgcttct gcatctcgtt gatcattttc tgggcgtcct
6841 tggagttctt ctcgcgggcc agctcgataa tgatgtcgtt gggcaggccg tacttcttga
6901 tgatggcgtt gatcactttg atgctctgga tgaagcttct cttcacgacg gggctcagga
6961 tgaagtcgtc caccagggtg gtggggatct ctttctgctg ggacaggtcc accttcttgg
7021 gcaccagctt cagccggttg aagatagcga tctggttgtc gttggtgtgc cacagctcgt
7081 ccaggatcag gttgatggcc ttcaggctca ggttgtgggt gccggtatag cccttcagat
7141 tagagatctg ctcgatctct tcctgggtca gctcggagtt cagattggtc agttcttcct
7201 ggatgtcctc gctgctctgg tagatggtca ggatcttggc aatctgatcc agcagctcgg
7261 cgttctcaat aatctctttc cgggcggtaa tgtccttgat gtcgtggtac accttcaggt
7321 tggtgaactc gggcttgccg gtgctggtca ctctgtagcc cttaatatcc tcttcgttca
7381 cgaggatttc tttggcgatc tgcttcaggg tgggcttctt cttctgcttg aacacgttct
7441 cgatga
//
