| 质粒名称: | pCMV-PEmax-P2A-BSD |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 174821 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV |
| 载体出品公司: | -- |
| 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: | 杀稻瘟菌素 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | PEmax-P2A-BSD,SV40_bpNLS-SpCas9(R221K,N394K,H840A)-SGGSx2-SV40_bpNLS-SGGSx2-MMLV_RT(D200N, T306K, W313F, T330P, L603W)-SV40_bpNLS-cMyc_NLS,-P2A-BSD |
| 物种来源: | Synthetic; SpCas9 is from Streptococcus pyogenes; MMLV_RT is from the Moloney murine leukemia virus |
| 插入大小: | 6855 bp |
| 基因突变: | Detailed in manuscript |
| GenBank编号 | -- |
| 启动子: | CMV |
| 标签/融合蛋白: |
SV40 bpNLS（插入件上的N端子）
SV40 bpNLS（插入件上的C端子）
c-Myc NLS（插入件上的c端子）
P2A-BSD（插入件上的C端子）
|
| 克隆方法: | Ligation Independent克隆 |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | T7-forward |
| 3’测序引物 | BGH-reverse |
|
文献引用方法 |
|
| 材料和方法部分: | pCMV-PEmax-P2A-BSD was a gift from David Liu (Addgene plasmid # 174821 ; http://n2t.net/addgene:174821 ; RRID:Addgene_174821) |
| 参考文献部分: | Enhanced prime editing systems by manipulating cellular determinants of editing outcomes. Chen PJ, Hussmann JA, Yan J, Knipping F, Ravisankar P, Chen PF, Chen C, Nelson JW, Newby GA, Sahin M, Osborn MJ, Weissman JS, Adamson B, Liu DR. Cell. 2021 Oct 9. pii: S0092-8674(21)01065-5. doi: 10.1016/j.cell.2021.09.018. 10.1016/j.cell.2021.09.018 PubMed 34653350 |
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 pCMV-PEmax-P2A-BSD 10257 bp ds-DNA circular SYN 27-SEP-2021
DEFINITION Mammalian expression of SpCas9 PEmax prime editor with P2A-BSD
marker.
ACCESSION .
VERSION .
KEYWORDS pCMV-PEmax-P2A-BSD
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10257)
TITLE Enhanced Prime Editing Systems Through Identification and
Manipulation of Cellular Determinants of Editing Outcomes
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 10257)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 27, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10257
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 12..391
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 392..595
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 546..566
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 637..656
/label=T7
/note="T7 promoter, forward primer"
promoter 637..655
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 705..725
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 726..4826
/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
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRKLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLKREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 4884..4904
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7005..7025
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7035..7061
/codon_start=1
/product="nuclear localization signal of human c-Myc
proto-oncogene (Dang and Lee, 1988)"
/label=c-myc NLS
/translation="PAAKRVKLD"
CDS 7071..7127
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 7128..7526
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
CDS 7532..7549
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(7572..7589)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 7578..7802
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(7873..7889)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7873..7889)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7886..7908)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7897..7913
/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)."
promoter complement(7921..7951)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7966..7987
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8104..8121)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8275..8863)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8355..8374)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(9034..9894)
/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 9657..9676
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9895..9999)
/gene="bla"
/label=AmpR promoter
primer_bind complement(10078..10097)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
61 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
121 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
181 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
241 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
301 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
361 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
421 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
481 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
541 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
601 agtgaaccgt cagatccgct agagatccgc ggccgctaat acgactcact atagggagag
661 ccgccaccat gaaacggaca gccgacggaa gcgagttcga gtcaccaaag aagaagcgga
721 aagtcgacaa gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg
781 tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc
841 ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg
901 aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct
961 gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca
1021 gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg
1081 gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa
1141 agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc
1201 acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg
1261 acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc
1321 ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca
1381 gaaagctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa
1441 acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg
1501 aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg
1561 cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca
1621 tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct
1681 ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc
1741 ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg
1801 ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc
1861 tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaagagagag gacctgctgc
1921 ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc
1981 acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga
2041 tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca
2101 gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg
2161 aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata
2221 agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg
2281 tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc
2341 tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga
2401 ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa
2461 tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa
2521 ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg
2581 tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg
2641 cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg
2701 gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc
2761 tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg
2821 acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc
2881 tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga
2941 cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg
3001 tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga
3061 gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc
3121 ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc
3181 gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacg
3241 ctatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa
3301 gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga
3361 agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc
3421 tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac
3481 agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga
3541 acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt
3601 ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca
3661 actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa
3721 agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga
3781 agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca
3841 gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc
3901 ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt
3961 ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg
4021 tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga
4081 tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg
4141 cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg
4201 tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg
4261 actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta
4321 agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac
4381 tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca
4441 gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg
4501 aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag
4561 tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata
4621 agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag
4681 cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca
4741 aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga
4801 tcgacctgtc tcagctggga ggtgactccg gcggaagctc tggtggcagc aagcggaccg
4861 ccgacggctc tgaattcgag agccctaaga agaaaagaaa ggtgagcgga ggctctagcg
4921 gcggaagcac cctgaacatt gaagacgagt atagactgca tgaaacaagc aaggaacccg
4981 acgtgtccct gggctccacc tggctgtccg actttcccca ggcctgggcc gagacaggag
5041 gaatgggcct ggccgtgcgg caggcacccc tgatcatccc tctgaaggcc acctctacac
5101 ccgtgagcat caagcagtac cctatgtctc aggaggccag actgggcatc aagcctcaca
5161 tccagaggct gctggaccag ggcatcctgg tgccatgcca gagcccctgg aacacaccac
5221 tgctgcccgt gaagaagcca ggcaccaatg actatagacc cgtgcaggat ctgagagagg
5281 tgaacaagag ggtggaggat atccacccca ccgtgcccaa cccttacaat ctgctgtccg
5341 gcctgccccc ttctcaccag tggtatacag tgctggacct gaaggatgcc ttcttttgtc
5401 tgagactgca ccctaccagc cagccactgt tcgcctttga gtggagggac cctgagatgg
5461 gcatctctgg ccagctgacc tggacacgcc tgcctcaggg cttcaagaat agcccaacac
5521 tgtttaacga ggccctgcac cgcgacctgg cagatttccg gatccagcac ccagatctga
5581 tcctgctgca gtacgtggac gatctgctgc tggccgccac cagcgagctg gattgccagc
5641 agggaacacg cgccctgctg cagaccctgg gaaacctggg atatagggca tccgccaaga
5701 aggcccagat ctgtcagaag caggtgaagt acctgggcta tctgctgaag gagggccaga
5761 gatggctgac agaggccagg aaggagacag tgatgggcca gccaacaccc aagaccccaa
5821 gacagctgag ggagttcctg ggcaaagcag gattttgcag gctgttcatc ccaggattcg
5881 cagagatggc agcacctctg tacccactga ccaagccggg caccctgttt aattggggcc
5941 ctgaccagca gaaggcctat caggagatca agcaggccct gctgacagca ccagccctgg
6001 gcctgccaga cctgaccaag cctttcgagc tgtttgtgga tgagaagcag ggctacgcca
6061 agggcgtgct gacccagaag ctgggaccat ggagacggcc cgtggcctat ctgtccaaga
6121 agctggaccc agtggcagca ggatggccac catgcctgag gatggtggca gcaatcgccg
6181 tgctgacaaa ggatgccggc aagctgacca tgggacagcc actggtcatc ctggcaccac
6241 acgcagtgga ggccctggtg aagcagcctc cagatcgctg gctgtctaac gcccggatga
6301 cacactacca ggccctgctg ctggacaccg atcgcgtgca gtttggccct gtggtggccc
6361 tgaatccagc caccctgctg cctctgccag aggagggcct gcagcacaac tgtctggaca
6421 tcctggcaga ggcacacgga acaaggccag acctgaccga tcagcccctg cctgacgccg
6481 atcacacatg gtataccgat ggaagctccc tgctgcagga gggccagagg aaggcaggag
6541 cagcagtgac cacagagaca gaagtgatct gggccaaggc cctgccagca ggcacatccg
6601 cccagcgggc cgagctgatc gccctgaccc aggccctgaa gatggccgag ggcaagaagc
6661 tgaacgtgta cacagactcc agatatgcct tcgccaccgc acacatccac ggagagatct
6721 acaggcgccg gggctggctg acctctgagg gcaaggagat caagaacaag gatgagatcc
6781 tggccctgct gaaggccctg tttctgccca agcggctgag catcatccac tgtcctggac
6841 accagaaggg acactccgcc gaggcaaggg gcaatcggat ggccgaccag gccgccagaa
6901 aggctgctat tactgaaact cccgacactt ccactctgct gattgaaaac tcctcccctt
6961 ctggcggctc aaaaagaacc gccgacggca gcgaattcga gtctcccaag aagaagagga
7021 aagtcggctc tggccctgcc gctaagagag tgaagctgga cggatccggc gcaacaaact
7081 tctctctgct gaaacaagcc ggagatgtcg aagagaatcc tggaccgatg gccaagcctt
7141 tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca
7201 tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc atcttcactg
7261 gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg
7321 ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg
7381 gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat cctgggatca
7441 aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc
7501 cctctggtta tgtgtgggag ggctaaccgg tcatcatcac catcaccatt gagtttaaac
7561 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
7621 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagaa
7681 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
7741 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
7801 ggcttctgag gcggaaagaa ccagctgggg ctcgataccg tcgacctcta gctagagctt
7861 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
7921 caacatacga gccggaagca taaagtgtaa agcctagggt gcctaatgag tgagctaact
7981 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
8041 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
8101 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
8161 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
8221 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
8281 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
8341 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
8401 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
8461 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
8521 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
8581 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
8641 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
8701 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
8761 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
8821 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
8881 ttctacgggg tctgacactc agtggaacga aaactcacgt taagggattt tggtcatgag
8941 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
9001 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
9061 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
9121 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
9181 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
9241 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
9301 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
9361 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
9421 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
9481 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
9541 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
9601 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
9661 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
9721 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
9781 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
9841 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
9901 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
9961 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
10021 acctgacgtc gacggatcgg gagatcgatc tcccgatccc ctagggtcga ctctcagtac
10081 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
10141 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
10201 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccag
//
