| 质粒名称: | pCMV-T7-SpRY-HF1-P2A-EGFP (RTW5008) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 139997 |
| 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-HF1 with BPNLS-3xFLAG-P2A-EGFP,RTW5008 |
| 物种来源: | Synthetic |
| 基因突变: | SpRY=A61R/L1111R/D1135L/S1136W/G1218K/E1219Q/N1317R/A1322R/R1333P/R1335Q/T1337R; HF1=N497A/R661A/Q695A/Q926A |
| 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-HF1-P2A-EGFP (RTW5008) was a gift from Benjamin Kleinstiver (Addgene plasmid # 139997 ; http://n2t.net/addgene:139997 ; RRID:Addgene_139997) |
| 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-HF1-P2A-EGFP (RTW5008) 8451 bp ds-DNA circular SYN 20-MAR-2020
DEFINITION CMV and T7 promoter expression plasmid for human codon optimized
SpRY-HF1(SpRY=A61R/L1111R/D1135L/S1136W/G1218K/E1219Q/N1317R/A1322R/
R1333P/R1335Q/T1337R;HF1=N497A/R661A/Q695A/Q926A) with a c-terminal
bi-partite NLS, 3x flag tag, and P2A-EGFP.
ACCESSION .
VERSION .
KEYWORDS pCMV-T7-SpRY-HF1-P2A-EGFP (RTW5008)
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JOURNAL unpublished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"
enhancer 165..544
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 545..748
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 699..719
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 790..809
/label=T7
/note="T7 promoter, forward primer"
promoter 790..808
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 816..825
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 822..2540
/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
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTAFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
CDS 4980..5000
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5013..5078
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 5097..5153
/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 5154..5873
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(5199..5220)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(5460..5479)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 5807..5828
/label=EGFP-C
/note="EGFP, forward primer"
CDS 5879..5896
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(5919..5936)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5925..6149
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(6220..6236)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6220..6236)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6233..6255)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6244..6260
/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(6268..6298)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6313..6334
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6451..6468)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6622..7210)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6702..6721)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7381..8241)
/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 8004..8023
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8242..8346)
/gene="bla"
/label=AmpR promoter
primer_bind complement(8425..8444)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
61 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
121 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
181 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
241 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
301 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
361 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
421 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
481 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
541 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
601 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
661 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
721 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagagatc
781 cgcggccgct aatacgactc actataggga gagccgccac catggacaag aagtacagca
841 tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg
901 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc
961 tgatcggagc cctgctgttc gacagcggcg aaacagccga gagaacccgg ctgaagagaa
1021 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca
1081 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg
1141 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg
1201 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg
1261 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc
1321 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc
1381 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg
1441 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg
1501 cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg
1561 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga
1621 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg
1681 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga
1741 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg
1801 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt
1861 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag
1921 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg
1981 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca
2041 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg
2101 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc
2161 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca
2221 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt
2281 ccgcccagag cttcatcgag cggatgaccg ccttcgataa gaacctgccc aacgagaagg
2341 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag
2401 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg
2461 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg
2521 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt
2581 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc
2641 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg
2701 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag
2761 tgatgaagca gctgaagcgg cggagataca ccggctgggg cgccctgagc cggaagctga
2821 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg
2881 gcttcgccaa cagaaacttc atggccctga tccacgacga cagcctgacc tttaaagagg
2941 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc
3001 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc
3061 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga
3121 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg
3181 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc
3241 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg
3301 aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc
3361 tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga
3421 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc
3481 tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg
3541 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccgggcca
3601 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg
3661 acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc
3721 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg
3781 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg
3841 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc
3901 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca
3961 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg
4021 gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc
4081 tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca
4141 aagagtctat cagacccaag aggaacagcg ataagctgat cgccagaaag aaggactggg
4201 accctaagaa gtacggcggc ttcctgtggc ccaccgtggc ctattctgtg ctggtggtgg
4261 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca
4321 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct
4381 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg
4441 aaaacggccg gaagagaatg ctggcctctg ccaagcagct gcagaaggga aacgaactgg
4501 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg
4561 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg
4621 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc
4681 tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg
4741 agaatatcat ccacctgttt accctgacca gactgggagc ccctagagcc ttcaagtact
4801 ttgacaccac catcgacccc aagcagtaca gaagcaccaa agaggtgctg gacgccaccc
4861 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag
4921 gcgacggatc cggcggaggc ggaagcggga aaagaaccgc cgacggcagc gaattcgagc
4981 ccaagaagaa gaggaaagtc tcgagcggag gcgactacaa agaccatgac ggtgattata
5041 aagatcatga catcgattac aaggatgacg atgacaaggg aggcggtgga agcggcgcaa
5101 caaacttctc tctgctgaaa caagccggag atgtcgaaga gaatcctgga ccgatggtga
5161 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
5221 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
5281 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
5341 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
5401 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
5461 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
5521 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
5581 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
5641 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
5701 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
5761 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
5821 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag tgaccggtca
5881 tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
5941 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
6001 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
6061 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
6121 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
6181 gataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
6241 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
6301 ctagggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
6361 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
6421 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
6481 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
6541 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
6601 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
6661 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
6721 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
6781 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
6841 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
6901 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
6961 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
7021 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
7081 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
7141 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
7201 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
7261 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
7321 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
7381 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
7441 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
7501 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
7561 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
7621 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
7681 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
7741 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
7801 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
7861 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
7921 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
7981 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
8041 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
8101 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
8161 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
8221 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
8281 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
8341 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcgatctcc
8401 cgatccccta gggtcgactc tcagtacaat ctgctctgat gccgcatagt t
//
