| 质粒名称: | pCMV-T7-SpRY-P2A-EGFP (RTW4830) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 139989 |
| 存储实验室: | Benjamin Kleinstiver |
|
载体骨架基本信息 |
|
| 载体名称: | RTW3027 |
| 载体出品公司: | Kleinstiver Lab (爱迪基因 ID 139987) |
| 载体抗性: | 氨苄青霉素 |
| 载体类型: | 哺乳动物表达，CRISPR；体外转录；T7启动子 |
| 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: | human codon optimized SpCas9 variant named SpRY with BPNLS-3xFLAG-P2A-EGFP,RTW4830 |
| 物种来源: | Synthetic |
| 基因突变: | SpRY=A61R/L1111R/D1135L/S1136W/G1218K/ E1219Q/N1317R/A1322R/R1333P/R1335Q/T1337R |
| 启动子 | CMV and T7 |
| 标签/融合蛋白: | BPNLS-3XLAGG-P2A-EGFP（插入件上的C端子） |
| 克隆方法: | Gibson 克隆 |
| 5' 测序引物 | oBK6928-CCAAGTCTCCACCCCATTGACG |
| 3' 测序引物 | oBK219-GGGAGTGGCACCTTCCAGGGTC |
|
文献引用方法 |
|
| 材料和方法部分: | pCMV-T7-SpRY-P2A-EGFP (RTW4830) was a gift from Benjamin Kleinstiver (Addgene plasmid # 139989 ; http://n2t.net/addgene:139989 ; RRID:Addgene_139989) |
| 参考文献部分: | Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants. Walton , Russell T., Christie, Kathleen A., Whittaker, Madelynn N., Kleinstiver, Benjamin P.. Science , 26 Mar 2020 10.1126/science.aba8853 |
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
质粒图谱.png)
载体简介
载体序列
LOCUS pCMV-T7-SpRY-P2A-EGFP (RTW4830) 8451 bp ds-DNA circular SYN 20-MAR-2020
DEFINITION CMV and T7 promoter expression plasmid for human codon optimized
SpRY(A61R/L1111R/D1135L/S1136W/G1218K/E1219Q/N1317R/A1322R/R1333P/R1
335Q/T1337R) with a c-terminal bi-partite NLS, 3x flag tag, and
P2A-EGFP.
ACCESSION .
VERSION .
KEYWORDS pCMV-T7-SpRY-P2A-EGFP (RTW4830)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8451)
AUTHORS Kleinstiver
TITLE Kleinstiver lab CRISPR plasmids
JOURNAL unpublished
REFERENCE 2 (bases 1 to 8451)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 20, 2020 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8451
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(250..1110)
/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 873..892
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1111..1215)
/gene="bla"
/label=AmpR promoter
primer_bind complement(1294..1313)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 1485..1864
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1865..2068
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2019..2039
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 2110..2129
/label=T7
/note="T7 promoter, forward primer"
promoter 2110..2128
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 2136..2145
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2142..3860
/codon_start=1
/product="N-terminal portion of Streptococcus pyogenes Cas9
(Zetsche et al., 2015)"
/label=Cas9(N)
/translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAERTRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
CDS 6300..6320
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6333..6398
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 6417..6473
/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 6474..7193
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(6519..6540)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(6780..6799)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 7127..7148
/label=EGFP-C
/note="EGFP, forward primer"
CDS 7199..7216
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(7239..7256)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 7245..7469
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(7540..7556)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7540..7556)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7553..7575)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7564..7580
/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(7588..7618)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7633..7654
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7771..7788)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7942..79)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8022..8041)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
61 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
121 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
181 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
241 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
301 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
361 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
421 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
481 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
541 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
601 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
661 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
721 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
781 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
841 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
901 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
961 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
1021 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
1081 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
1141 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
1201 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
1261 tcgatctccc gatcccctag ggtcgactct cagtacaatc tgctctgatg ccgcatagtt
1321 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
1381 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
1441 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
1501 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
1561 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
1621 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
1681 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
1741 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
1801 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
1861 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
1921 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
1981 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
2041 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagagatc
2101 cgcggccgct aatacgactc actataggga gagccgccac catggacaag aagtacagca
2161 tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg
2221 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc
2281 tgatcggagc cctgctgttc gacagcggcg aaacagccga gagaacccgg ctgaagagaa
2341 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca
2401 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg
2461 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg
2521 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg
2581 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc
2641 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc
2701 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg
2761 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg
2821 cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg
2881 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga
2941 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg
3001 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga
3061 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg
3121 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt
3181 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag
3241 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg
3301 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca
3361 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg
3421 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc
3481 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca
3541 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt
3601 ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg
3661 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag
3721 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg
3781 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg
3841 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt
3901 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc
3961 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg
4021 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag
4081 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga
4141 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg
4201 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg
4261 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc
4321 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc
4381 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga
4441 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg
4501 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc
4561 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg
4621 aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc
4681 tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga
4741 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc
4801 tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg
4861 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga
4921 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg
4981 acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc
5041 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg
5101 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg
5161 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc
5221 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca
5281 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg
5341 gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc
5401 tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca
5461 aagagtctat cagacccaag aggaacagcg ataagctgat cgccagaaag aaggactggg
5521 accctaagaa gtacggcggc ttcctgtggc ccaccgtggc ctattctgtg ctggtggtgg
5581 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca
5641 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct
5701 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg
5761 aaaacggccg gaagagaatg ctggcctctg ccaagcagct gcagaaggga aacgaactgg
5821 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg
5881 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg
5941 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc
6001 tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg
6061 agaatatcat ccacctgttt accctgacca gactgggagc ccctagagcc ttcaagtact
6121 ttgacaccac catcgacccc aagcagtaca gaagcaccaa agaggtgctg gacgccaccc
6181 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag
6241 gcgacggatc cggcggaggc ggaagcggga aaagaaccgc cgacggcagc gaattcgagc
6301 ccaagaagaa gaggaaagtc tcgagcggag gcgactacaa agaccatgac ggtgattata
6361 aagatcatga catcgattac aaggatgacg atgacaaggg aggcggtgga agcggcgcaa
6421 caaacttctc tctgctgaaa caagccggag atgtcgaaga gaatcctgga ccgatggtga
6481 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
6541 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
6601 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
6661 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
6721 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
6781 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
6841 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
6901 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
6961 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
7021 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
7081 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
7141 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag tgaccggtca
7201 tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
7261 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
7321 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
7381 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
7441 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
7501 gataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
7561 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
7621 ctagggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
7681 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
7741 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
7801 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
7861 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
7921 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
7981 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
8041 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
8101 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
8161 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
8221 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
8281 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
8341 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
8401 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat c
//
