| 质粒名称: | pXR002: EF1a-dCasRx-2A-EGFP |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 74286 |
| 存储实验室: | Patrick Hsu |
|
载体骨架基本信息 |
|
| 载体名称: | lenti(AMP) |
| 完整质粒大小: | 12990bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | GFP |
| 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: | dCasRx/NLS-dRfxCas13d-NLS |
| 物种来源: | H. sapiens (human); Ruminococcus flavefaciens XPD3002 |
| 插入大小: | 3000 bp |
| 基因突变: | R239A/H244A/R858A/H863A |
| 标签/融合蛋白: |
NLS (N terminal on insert)
NLS (C terminal on insert)
HA (C terminal on insert)
2A-eGFP (C terminal on backbone)
|
|
文献引用方法 |
|
| 材料和方法部分: | pXR002: EF1a-dCasRx-2A-EGFP was a gift from Patrick Hsu (Addgene plasmid # 109050 ; http://n2t.net/addgene:109050 ; RRID:Addgene_109050) |
| 参考文献部分: | Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR Effectors. Konermann S, Lotfy P, Brideau NJ, Oki J, Shokhirev MN, Hsu PD. Cell. 2018 Mar 8. pii: S0092-8674(18)30207-1. doi: 10.1016/j.cell.2018.02.033. 10.1016/j.cell.2018.02.033 PubMed 29551272 |
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 pXR002: EF1a-dCasRx-2A-EGFP 12991 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Catalytically inactive dCasRx with 2A-EGFP.
ACCESSION .
VERSION .
KEYWORDS pXR002: EF1a-dCasRx-2A-EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12991)
AUTHORS Konermann S, Lotfy P, Brideau NJ, Oki J, Shokhirev MN, Hsu PD
TITLE Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR
Effectors.
JOURNAL Cell. 2018 Mar 8. pii: S0092-8674(18)30207-1. doi:
10.1016/j.cell.2018.02.033.
PUBMED 29551272
REFERENCE 2 (bases 1 to 12991)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..12991
/organism="synthetic DNA construct"
/mol_type="other DNA"
polyA_signal 60..284
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 273..290
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
LTR 316..496
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind 1001..1017
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 1002..1018
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 1021..1609
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS 1127..1138
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind 1536..1556
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(1634..2350)
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
CDS complement(1637..2350)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(1679..1700)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 2028..2047
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2287..2308
/label=EGFP-N
/note="EGFP, reverse primer"
CDS complement(2351..2404)
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS complement(2420..2446)
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS complement(2456..2476)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS complement(2486..5383)
/codon_start=1
/product="RNA-targeting Type VI CRISPR protein from
Ruminococcus flavefaciens strain XPD3002 (Konermann et al.,
2018)"
/label=RfxCas13d
/translation="IEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDSI
RSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLEK
RYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFSTVY
TYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYIIN
YGNECYDILALLSGLAHWVVANNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNE
LTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKD
MSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVI
NLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPAG
RDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAFF
KDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDEN
GNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQK
KQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGREN
AEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLEE
KGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKKA
EEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFANKAVALEVARYVHA
YINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVP
FGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS"
CDS complement(5393..5413)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
promoter complement(5432..6610)
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 5441..6379
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind complement(5464..5484)
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
misc_feature 6735..6852
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(7150..7194)
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 7379..7612
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 8105..8230
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
LTR 8277..8457
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(8475..8673)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(8500..8520)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 8675..9054
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind 9226..9245
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 9320..9424
/gene="bla"
/label=AmpR promoter
CDS 9425..10285
/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 complement(9643..9662)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 10456..11044
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 10945..10964
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 11198..11215
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 11332..11353
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 11368..11398
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 11406..11422
/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)."
primer_bind 11411..11433
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 11430..11446
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 11430..11446
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
polyA_signal 11495..11616
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(11506..11525)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 11560..11579
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(11746..12120)
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
promoter complement(12139..12186)
/label=EM7 promoter
/note="synthetic bacterial promoter "
promoter complement(12234..12563)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 12248..12383
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(12302..12321)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 12548..12568
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin complement(12577..14)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(12687..12708)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 12899..12918
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 gctacagggc gcgtggggat accccctaga gccccagctg gttctttccg cctcagaagc
61 catagagccc accgcatccc cagcatgcct gctattgtct tcccaatcct cccccttgct
121 gtcctgcccc accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat
181 ttcctcattt tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac
241 gggggagggg caaacaacag atggctggca actagaaggc acagtcgagg ctgatcagcg
301 ggtttaaacg ggccctgcta gagattttcc acactgacta aaagggtctg agggatctct
361 agttaccaga gtcacacaac agacgggcac acactacttg aagcactcaa ggcaagcttt
421 attgaggctt aagcagtggg ttccctagtt agccagagag ctcccaggct cagatctggt
481 ctaaccagag agacccagta cagtccggat gcagctctcg ggccatgtga tgaaatgcta
541 ggcggctgtc aaacctccac tctaatactt ctctctccgg gtcatccatc ccatgcaggc
601 tcacagggtg taacaagcgg gtgttctctc cttcattggc ttcttctacc ttctcttgct
661 caactggtac tagcttgtag caccatccaa aggtcagtgg atatctgatc cctggccctg
721 gtgtgtagtt ctgccaatca gggaagtagc cttgtgtgtg gtagatccac agatcaagga
781 tatcttgtct tcgttgggag tgaattagcc cttccagtcc ccccttttct tttaaaaagt
841 ggctaagatc tacagctgcc ttgtaagtca ttggtcttaa aggtacctga ggtgtgactg
901 gaaaacccac ctcctcctcc tcttgtgctt ctagccaggc acaatcagca ttggtagctg
961 ctgtattgct acttgtgatt gctccatgtt tttctaggtc tcgaggtcga cggtatcgat
1021 gcggggaggc ggcccaaagg gagatccgac tcgtctgagg gcgaaggcga agacgcggaa
1081 gaggccgcag agccggcagc aggccgcggg aaggaaggtc cgctggattg agggccgaag
1141 ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc aacacaggcg agcagccaag
1201 gaaaggacga tgatttcccc gacaacacca cggaattgtc agtgcccaac agccgagccc
1261 ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaata gggaggggga
1321 aagcgaaagt cccggaaagg agctgacagg tggtggcaat gccccaacca gtgggggttg
1381 cgtcagcaaa cacagtgcac accacgccac gttgcctgac aacgggccac aactcctcat
1441 aaagagacag caaccaggat ttatacaagg aggagaaaat gaaagccata cgggaagcaa
1501 tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga gcaacatagt
1561 taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgatta tcgataagct
1621 tgatatcgaa ttcttacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
1681 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact
1741 gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
1801 gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt
1861 tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt
1921 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
1981 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
2041 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
2101 agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca
2161 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca
2221 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
2281 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
2341 ccttgctcac tgggccagga ttctcctcga cgtcaccgca tgttagcaga cttcctctgc
2401 cctctccact gccgctagca gcgtaatctg gaacatcgta tgggtaagcg gccgccacct
2461 tcctcttttt cttaggtccg gatccggaat tgccggacac cttctttttc tccttgtcga
2521 acttggcggc ctcgttccta tcgaacaggg cctcgatgct caggttctta aacctgggga
2581 tacagtagcc gaaaggcaca cacagcagtt tcaggagcct atcgttgtac ttcttctcgt
2641 cattcacagc gtcgaagtac tcggacacct ttccgctgct tttctcgtac ctctcattca
2701 tgataattct ctgcatgatg taatggtaca gttggaagta ggaattgacc tcggcaatgt
2761 cgttgatata ggcgtggaca tacctggcca cttccagggc gacggccttg tttgcgaaca
2821 gggtacatgt cttgttgtca attctcagga ggtcctccct gatgatcaca ttccacttgg
2881 tgttcctcag gtaggcgttc agggcggttt tggccttctc cctgttaatc tgcctggtga
2941 actcctcggc tttcttctcg tcgctgtatt tgatgtaatt ggcatacagc ttggggttgg
3001 cgctctcgag gctgtcaatg ctctccttgg ctctttcagc catctccttt tccacgtcct
3061 ttctcttatc gggggcagtt tcatcaatgc cagcgcagag cttggtgacg gagctgaatc
3121 ccttctcttc cagtttcttg agattgatgt cgtagccttt ctccttgtac agttgagcat
3181 cacgctcgac gcaatggaat ccgatgacgt acctggcgtt gatattgaca atattcttga
3241 ggatgtggta gatcacggtg aggtacaggc tgatgatctt tttaaacttc tccctctcgg
3301 cgttttccct gccggtgtcc tcaatgacgc tccttttctt gtcgaattgg tcgtagttca
3361 ttccggtgat gatctttgtg agagcgtcca ccttttcgct cacgctcttg cccttatcct
3421 ttccgataca agtttcgtag tacctgtcga tctggttctt gccgttctgg ccctgttttt
3481 tctggatgtc agcgatcctg ccgagcacga acttcaccac ggcctcgttt ttggcgatct
3541 catggaggtg ggcaggatca ccgtatctga tcaggtagtg gaacctttta ttgctgatca
3601 cgttattaat aatgaaattt ctcatgccgt gcttgccttt cttgagcttg tttccgttct
3661 cgtccaggga aaaggtgtcg gcgagggcct tgagctcatc ataggacagg ttggttccta
3721 aaatacggat ggcgtcgata tacatggccc tcctggcatc ggcaataggt tctcccattc
3781 tagcgaagga cttgatcagc ctcagctcat cggcgatctt ggcggagtct ttgaaaaagg
3841 cgtattcctc cacgaactta gcgttgactc cgatgagagg catcaccttc aggaagctct
3901 ggatgttatc gaatttatta atcagggtgg tcaggaggtc gttgatctcc ttgccatcca
3961 ggaacatggt cagggcatac atgagtttgc tgaaggcgga aacatcacgg ccagcgggca
4021 ggattctggg cagtctaggg gcgtccttct tcttatactc tcttgtcttg tttcccctaa
4081 attccttgat gttgtgcatg atattttcga gctttctcca aattctatta gcttcatcgt
4141 agtagagggc atccttctgg tcgtcgttga aggagcccct caggttaatc acaaagatat
4201 ccttctcgct cagggacttc tcattatcgg ggagggactt attggcggca gccaccttgg
4261 catcctcttc gatgtaatac ctataaatca caaagtccat catggtgtag accttggtcc
4321 tgatggagtc gaacacctta tgatttttcc tgatctcgga catatccttc ctgtccagca
4381 tcacttccct gagcttggtg atattgaatc cgaggttttt ctgctctttc ataatgctga
4441 atctgaaata ttgttcggcg aattcggcag ggttgattcc cagagtttcg gcaatatagt
4501 tcacgttggc ggcggagttc ttggagaagg agttggtcag ctcattggtg atcctgtcgt
4561 agaggtagtt gagggtggag atgtattcgt tgtcgaggtt cttatcgagg ttgtagagcc
4621 aggtcctgga gatcctggac tcttcttcgt tgttagcgac cacccagtgc gccagtccgc
4681 tcaggagggc cagaatgtca tagcattcgt tgccgtaatt gatgatgtaa tttctgccct
4741 ccttgctgaa aaaggcctgg ccgaaatagc cgagtctggg gttatcgagg aagttgtcga
4801 actcgtcata ctgggccttg atggcgttga tgagcttatc gttattgttg aaagcggccc
4861 tatggtgctc ggggtctttg aattcgtcgt aggtatacac tgtggagaac ttgccgaatc
4921 caataatgtc cttatccagg ccggagatat tgttgacggc gtaggcggcg ttggtaatgt
4981 attcggcgag gattttttca atgtccagga tgttatggat cacctggata caaatattgt
5041 cattgccatc agcgctctcg ccgaagtacc tcttttccag agtttccttc aggccgagca
5101 tatcctgctg gacgggtcct gtatacagag ggttgttagc caccacggcg tagcccttag
5161 gatggctgaa tttggcgttg ccgatcttat agccggcgtt tttatcggcc atttcagcgc
5221 tgaaggcctc gccctcattc acgctcctga tgctgtcgcc ctccacgatc ttttccagcc
5281 tggcgtcgct gccttcggcg aaggttgtca tgtacacttt ggagccggac acgagtgtgg
5341 acttcacgcc catgcccttg gcgaaggact tttttttttc gatgctggcc tccacctttc
5401 tcttcttctt ggggctcatg gtggccgtac gtcacgacac ctgaaatgga agaaaaaaac
5461 tttgaaccac tgtctgaggc ttgagaatga accaagatcc aaactcaaaa agggcaaatt
5521 ccaaggagaa ttacatcaag tgccaagctg gcctaacttc agtctccacc cactcagtgt
5581 ggggaaactc catcgcataa aacccctccc cccaacctaa agacgacgta ctccaaaagc
5641 tcgagaacta atcgaggtgc ctggacggcg cccggtactc cgtggagtca catgaagcga
5701 cggctgagga cggaaaggcc cttttccttt gtgtgggtga ctcacccgcc cgctctcccg
5761 agcgccgcgt cctccatttt gagctccctg cagcagggcc gggaagcggc catctttccg
5821 ctcacgcaac tggtgccgac cgggccagcc ttgccgccca gggcggggcg atacacggcg
5881 gcgcgaggcc aggcaccaga gcaggccggc cagcttgaga ctacccccgt ccgattctcg
5941 gtggccgcgc tcgcaggccc cgcctcgccg aacatgtgcg ctgggacgca cgggccccgt
6001 cgccgcccgc ggccccaaaa accgaaatac cagtgtgcag atcttggccc gcatttacaa
6061 gactatcttg ccagaaaaaa agcgtcgcag caggtcatca aaaattttaa atggctagag
6121 acttatcgaa agcagcgaga caggcgcgaa ggtgccacca gattcgcacg cggcggcccc
6181 agcgcccagg ccaggcctca actcaagcac gaggcgaagg ggctccttaa gcgcaaggcc
6241 tcgaactctc ccacccactt ccaacccgaa gctcgggatc aagaatcacg tactgcagcc
6301 aggtggaagt aattcaaggc acgcaagggc cataacccgt aaagaggcca ggcccgcggg
6361 aaccacacac ggcacttacc tgtgttctgg cggcaaaccc gttgcgaaaa agaacgttca
6421 cggcgactac tgcacttata tacggttctc ccccaccctc gggaaaaagg cggagccagt
6481 acacgacatc actttcccag tttaccccgc gccaccttct ctaggcaccg gttcaattgc
6541 cgacccctcc ccccaacttc tcggggactg tgggcgatgt gcgctctgcc cactgacggg
6601 caccggagcc aattcccact cctttcaaga cctagaaggt ccattagctg caaagattcc
6661 tctctgttta aaactttatc catctttgca ttaattaacc aaactggatc tctgctgtcc
6721 ctgtaataaa cccgaaaatt ttgaattttt gtaatttgtt tttgtaattc tttagtttgt
6781 atgtctgttg ctattatgtc tactattctt tcccctgcac tgtacccccc aatcccccct
6841 tttcttttaa aattgtggat gaatactgcc atttgtctgc agaattggcg cacgcagtgc
6901 cgatccgttc actaatcgaa tggatctgtc tctgtctctc tctccacctt cttcttctat
6961 tccttcgggc ctgtcgggtc ccctcggggt tgggaggtgg gtctgaaacg ataatggtga
7021 atatccctgc ctaactctat tcactataga aagtacagca aaaactattc ttaaacctac
7081 caagcctcct actatcatta tgaataattt tatataccac agccaatttg ttatgttaaa
7141 ccaattccac aaacttgccc atttatctaa ttccaataat tcttgttcat tcttttcttg
7201 ctggttttgc gattcttcaa ttaaggagtg tattaagctt gtgtaattgt taatttctct
7261 gtcccactcc atccaggtcg tgtgattcca aatctgttcc agagatttat tactccaact
7321 agcattccaa ggcacagcag tggtgcaaat gagttttcca gagcaacccc aaatccccag
7381 gagctgttga tcctttaggt atctttccac agccaggatt cttgcctgga gctgcttgat
7441 gccccagact gtgagttgca acagatgctg ttgcgcctca atagccctca gcaaattgtt
7501 ctgctgctgc actataccag acaataattg tctggcctgt accgtcagcg tcattgacgc
7561 tgcgcccata gtgcttcctg ctgctcccaa gaacccaagg aacaaagctc ctattcccac
7621 tgctcttttt tctctctgca ccactcttct ctttgccttg gtgggtgcta ctcctaatgg
7681 ttcaattttt actactttat atttatataa ttcacttctc caattgtccc tcatatctcc
7741 tcctccaggt ctgaagatca gcggccgctt gctgtgcggt ggtcttactt ttgttttgct
7801 cttcctctat cttgtctaaa gcttccttgg tgtcttttat ctctatcctt tgatgcacac
7861 aatagagggt tgctactgta ttatataatg atctaagttc ttctgatcct gtctgaaggg
7921 atggttgtag ctgtcccagt atttgtctac agccttctga tgtttctaac aggccaggat
7981 taactgcgaa tcgttctagc tccctgcttg cccatactat atgttttaat ttatattttt
8041 tctttccccc tggccttaac cgaatttttt cccatcgcga tctaattctc ccccgcttaa
8101 tactgacgct ctcgcaccca tctctctcct tctagcctcc gctagtcaaa atttttggcg
8161 tactcaccag tcgccgcccc tcgcctcttg ccgtgcgcgc ttcagcaagc cgagtcctgc
8221 gtcgagagag ctcctctggt ttccctttcg ctttcaagtc cctgttcggg cgccactgct
8281 agagattttc cacactgact aaaagggtct gagggatctc tagttaccag agtcacacaa
8341 cagacgggca cacactactt gaagcactca aggcaagctt tattgaggct taagcagtgg
8401 gttccctagt tagccagaga gctcccaggc tcagatctgg tctaaccaga gagacccagt
8461 acaggcaaaa cgcgctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg
8521 tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca
8581 aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc
8641 cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt
8701 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg
8761 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta
8821 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
8881 ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg
8941 gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat
9001 gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa tgtcaacgcg
9061 tatatctggc ccgtacatcg cgaagcagcg caaaacgcct aaccctaagc agattcttca
9121 tgcaattgtc ggtcaagcct tgccttgttg tagcttaaat tttgctcgcg cactactcag
9181 cgacctccaa cacacaagca gggagcagat actggcttaa ctatgcggca tcagagcaga
9241 ttgtactgag agtgcaccat aggggatcgg gagatctccc gatccgtcga cgtcaggtgg
9301 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
9361 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
9421 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
9481 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
9541 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
9601 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
9661 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
9721 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
9781 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
9841 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
9901 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
9961 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
10021 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
10081 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
10141 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
10201 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
10261 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
10321 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
10381 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
10441 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
10501 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
10561 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
10621 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
10681 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
10741 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
10801 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
10861 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
10921 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
10981 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
11041 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
11101 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
11161 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
11221 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
11281 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
11341 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
11401 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
11461 gctctagcta gaggtcgacg gtatacagac atgataagat acattgatga gtttggacaa
11521 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
11581 ttatttgtaa ccattataag ctgcaataaa caagttgggg tgggcgaaga actccagcat
11641 gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa
11701 cctttcatag aaggcggcgg tggaatcgaa atctcgtagc acgtgtcagt cctgctcctc
11761 ggccacgaag tgcacgcagt tgccggccgg gtcgcgcagg gcgaactccc gcccccacgg
11821 ctgctcgccg atctcggtca tggccggccc ggaggcgtcc cggaagttcg tggacacgac
11881 ctccgaccac tcggcgtaca gctcgtccag gccgcgcacc cacacccagg ccagggtgtt
11941 gtccggcacc acctggtcct ggaccgcgct gatgaacagg gtcacgtcgt cccggaccac
12001 accggcgaag tcgtcctcca cgaagtcccg ggagaacccg agccggtcgg tccagaactc
12061 gaccgctccg gcgacgtcgc gcgcggtgag caccggaacg gcactggtca acttggccat
12121 ggtttagttc ctcaccttgt cgtattatac tatgccgata tactatgccg atgattaatt
12181 gtcaacacgt gctgatcaga tccgaaaatg gatatacaag ctcccgggag ctttttgcaa
12241 aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggcagaggcg
12301 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
12361 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
12421 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
12481 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
12541 tttccacacc ctaactgaca cacattccac agaattaatt cgcgttaaat ttttgttaaa
12601 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
12661 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
12721 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
12781 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
12841 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
12901 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
12961 taaccaccac acccgccgcg cttaatgcgc c
//
