| 质粒名称: | pT3-EF1A-MYC-IRES-lucOS |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 129776 |
| 存储实验室: | Amaia Lujambio |
|
载体骨架基本信息 |
|
| 载体名称: | pT3-EF1a |
| 空载体大小: | 9597 bp |
| 完整质粒大小: | 9598bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | human MYC / MYC |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 4100 bp |
| 基因突变: | T2A, S13L, A92T, V104T, V114I, E161K, Y193K, F198Y, D202T, I203V, Y258F, A283V, K288T, N290Y relative to RpBphP2 |
| 启动子: | EF1A |
| 标签/融合蛋白: |
luciferase (C terminal on insert)
OS: SIYRYYGL (SIY), SIINFEKL (SIN, OVA257-264), and OVA323-339 (C terminal on insert)
|
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | TCAAGCCTCAGACAGTGGTTC |
|
文献引用方法 |
|
| 材料和方法部分: | pT3-EF1A-MYC-IRES-lucOS was a gift from Amaia Lujambio (Addgene plasmid # 129776 ; http://n2t.net/addgene:129776 ; RRID:Addgene_129776) |
| 参考文献部分: | beta-catenin activation promotes immune escape and resistance to anti-PD-1 therapy in hepatocellular carcinoma. Ruiz de Galarreta M, Bresnahan E, Molina-Sanchez P, Lindblad KE, Maier B, Sia D, Puigvehi M, Miguela V, Casanova-Acebes M, Dhainaut M, Villacorta-Martin C, Singhi AD, Moghe A, von Felden J, Tal Grinspan L, Wang S, Kamphorst AO, Monga SP, Brown BD, Villanueva A, Llovet JM, Merad M, Lujambio A. Cancer Discov. 2019 Jun 11. pii: 2159-8290.CD-19-0074. doi: 10.1158/2159-8290.CD-19-0074. 10.1158/2159-8290.CD-19-0074 PubMed 31186238 |
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 pT3-EF1A-MYC-IRES-lucOS 9598 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Transposon based vector that expresses human MYC followed by IRES
and firefly luciferase and the antigens SIYRYYGL (SIY), SIINFEKL
(SIN, OVA257-264), and OVA323-339.
ACCESSION .
VERSION .
KEYWORDS pT3-EF1A-MYC-IRES-lucOS
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9598)
AUTHORS Ruiz de Galarreta M, Bresnahan E, Molina-Sanchez P, Lindblad KE,
Maier B, Sia D, Puigvehi M, Miguela V, Casanova-Acebes M, Dhainaut
M, Villacorta-Martin C, Singhi AD, Moghe A, von Felden J, Tal
Grinspan L, Wang S, Kamphorst AO, Monga SP, Brown BD, Villanueva A,
Llovet JM, Merad M, Lujambio A
TITLE beta-catenin activation promotes immune escape and resistance to
anti-PD-1 therapy in hepatocellular carcinoma.
JOURNAL Cancer Discov. 2019 Jun 11. pii: 2159-8290.CD-19-0074. doi:
10.1158/2159-8290.CD-19-0074.
PUBMED 31186238
REFERENCE 2 (bases 1 to 9598)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9598
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind complement(34..67)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
promoter 208..1386
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 439..1377
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1334..1354
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
primer_bind 1403..1422
/label=T7
/note="T7 promoter, forward primer"
promoter 1403..1421
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1500..1524
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 1533..2849
/codon_start=1
/product="human c-Myc proto-oncogene"
/label=c-Myc
/translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP
SEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE
LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN
PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS
LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS
PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR
KCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA
TAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA"
protein_bind complement(2851..2875)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 2928..2969
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
misc_feature 2991..3555
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
misc_feature 3005..3557
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(3149..3166)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 3376..3395
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 3569..5218
/codon_start=1
/gene="luc+"
/product="firefly luciferase"
/label=luciferase
/note="enhanced luc+ version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
primer_bind complement(3635..3653)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
CDS 5231..5254
/codon_start=1
/product="antigenic CD8+ T cell epitope from chicken
ovalbumin"
/label=OVA peptide
/translation="SIINFEKL"
CDS 5672..5701
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
primer_bind complement(5727..5744)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5733..5957
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
protein_bind complement(6027..6060)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
primer_bind 6116..6136
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
primer_bind complement(6518..6535)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(6518..6534)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6527..6549)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(6743..6762)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6862..6884
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6922..6940)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 7008..7112
/gene="bla"
/label=AmpR promoter
CDS 7113..7973
/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(7331..7350)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8144..8732
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8633..8652
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8886..8903
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 9020..9041
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 9056..9086
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9094..9110
/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 9099..9121
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 9118..9134
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 9118..9134
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(9560..9579)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
ORIGIN
1 ctagctcgag atcttaatta aactagtatc gatataactt cgtataatgt atgctatacg
61 aagttatatg tacgggccag atatacgcgt tgacattgat tattgactag gcttttgcaa
121 aaagctttgc aaagatggat aaagttttaa acagagagga atctttgcag ctaatggacc
181 ttctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca
241 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga
301 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
361 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
421 tttgccgcca gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg
481 ggttatggcc cttgcgtgcc ttgaattact tccacctggc tgcagtacgt gattcttgat
541 cccgagcttc gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct
601 tcgcctcgtg cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg
661 tggcaccttc gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga
721 tgacctgctg cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg
781 cacactggta tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc
841 acatgttcgg cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct
901 caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg
961 gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc
1021 cctgctgcag ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca
1081 cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag
1141 taccgggcgc cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta
1201 ggttgggggg aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa
1261 gttaggccag cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga
1321 tcttggttca ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg
1381 tcgtgaggaa ttagcttggt actaatacga ctcactatag ggagacccaa gctggctagg
1441 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattctg cagatatcaa
1501 caagtttgta caaaaaagca ggcttcgcca ccatgcccct caacgttagc ttcaccaaca
1561 ggaactatga cctcgactac gactcggtgc agccgtattt ctactgcgac gaggaggaga
1621 acttctacca gcagcagcag cagagcgagc tgcagccccc ggcgcccagc gaggatatct
1681 ggaagaaatt cgagctgctg cccaccccgc ccctgtcccc tagccgccgc tccgggctct
1741 gctcgccctc ctacgttgcg gtcacaccct tctcccttcg gggagacaac gacggcggtg
1801 gcgggagctt ctccacggcc gaccagctgg agatggtgac cgagctgctg ggaggagaca
1861 tggtgaacca gagtttcatc tgcgacccgg acgacgagac cttcatcaaa aacatcatca
1921 tccaggactg tatgtggagc ggcttctcgg ccgccgccaa gctcgtctca gagaagctgg
1981 cctcctacca ggctgcgcgc aaagacagcg gcagcccgaa ccccgcccgc ggccacagcg
2041 tctgctccac ctccagcttg tacctgcagg atctgagcgc cgccgcctca gagtgcatcg
2101 acccctcggt ggtcttcccc taccctctca acgacagcag ctcgcccaag tcctgcgcct
2161 cgcaagactc cagcgccttc tctccgtcct cggattctct gctctcctcg acggagtcct
2221 ccccgcaggg cagccccgag cccctggtgc tccatgagga gacaccgccc accaccagca
2281 gcgactctga ggaggaacaa gaagatgagg aagaaatcga tgttgtttct gtggaaaaga
2341 ggcaggctcc tggcaaaagg tcagagtctg gatcaccttc tgctggaggc cacagcaaac
2401 ctcctcacag cccactggtc ctcaagaggt gccacgtctc cacacatcag cacaactacg
2461 cagcgcctcc ctccactcgg aaggactatc ctgctgccaa gagggtcaag ttggacagtg
2521 tcagagtcct gagacagatc agcaacaacc gaaaatgcac cagccccagg tcctcggaca
2581 ccgaggagaa tgtcaagagg cgaacacaca acgtcttgga gcgccagagg aggaacgagc
2641 taaaacggag cttttttgcc ctgcgtgacc agatcccgga gttggaaaac aatgaaaagg
2701 cccccaaggt agttatcctt aaaaaagcca cagcatacat cctgtccgtc caagcagagg
2761 agcaaaagct catttctgaa gaggacttgt tgcggaaacg acgagaacag ttgaaacaca
2821 aacttgaaca gctacggaac tcttgtgcgg acccagcttt cttgtacaaa gtggttgata
2881 tccagcacag tggcggccgc tcgagtctag agggcccgcg gttcgaaggt aagcctatcc
2941 ctaaccctct cctcggtctc gattctacgc gtaccggtta gtaatgagtt tccccccccc
3001 cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta
3061 ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc
3121 ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat
3181 gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc
3241 ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt
3301 gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt
3361 gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag
3421 aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt
3481 tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga
3541 aaaacacgat gataatatgg ccacaaccat ggaagacgcc aaaaacataa agaaaggccc
3601 ggcgccattc tatccgctgg aagatggaac cgctggagag caactgcata aggctatgaa
3661 gagatacgcc ctggttcctg gaacaattgc ttttacagat gcacatatcg aggtggacat
3721 cacttacgct gagtacttcg aaatgtccgt tcggttggca gaagctatga aacgatatgg
3781 gctgaataca aatcacagaa tcgtcgtatg cagtgaaaac tctcttcaat tctttatgcc
3841 ggtgttgggc gcgttattta tcggagttgc agttgcgccc gcgaacgaca tttataatga
3901 acgtgaattg ctcaacagta tgggcatttc gcagcctacc gtggtgttcg tttccaaaaa
3961 ggggttgcaa aaaattttga acgtgcaaaa aaagctccca atcatccaaa aaattattat
4021 catggattct aaaacggatt accagggatt tcagtcgatg tacacgttcg tcacatctca
4081 tctacctccc ggttttaatg aatacgattt tgtgccagag tccttcgata gggacaagac
4141 aattgcactg atcatgaact cctctggatc tactggtctg cctaaaggtg tcgctctgcc
4201 tcatagaact gcctgcgtga gattctcgca tgccagagat cctatttttg gcaatcaaat
4261 cattccggat actgcgattt taagtgttgt tccattccat cacggttttg gaatgtttac
4321 tacactcgga tatttgatat gtggatttcg agtcgtctta atgtatagat ttgaagaaga
4381 gctgtttctg aggagccttc aggattacaa gattcaaagt gcgctgctgg tgccaaccct
4441 attctccttc ttcgccaaaa gcactctgat tgacaaatac gatttatcta atttacacga
4501 aattgcttct ggtggcgctc ccctctctaa ggaagtcggg gaagcggttg ccaagaggtt
4561 ccatctgcca ggtatcaggc aaggatatgg gctcactgag actacatcag ctattctgat
4621 tacacccgag ggggatgata aaccgggcgc ggtcggtaaa gttgttccat tttttgaagc
4681 gaaggttgtg gatctggata ccgggaaaac gctgggcgtt aatcaaagag gcgaactgtg
4741 tgtgagaggt cctatgatta tgtccggtta tgtaaacaat ccggaagcga ccaacgcctt
4801 gattgacaag gatggatggc tacattctgg agacatagct tactgggacg aagacgaaca
4861 cttcttcatc gttgaccgcc tgaagtctct gattaagtac aaaggctatc aggtggctcc
4921 cgctgaattg gaatccatct tgctccaaca ccccaacatc ttcgacgcag gtgtcgcagg
4981 tcttcccgac gatgacgccg gtgaacttcc cgccgccgtt gttgttttgg agcacggaaa
5041 gacgatgacg gaaaaagaga tcgtggatta cgtcgccagt caagtaacaa ccgcgaaaaa
5101 gttgcgcgga ggagttgtgt ttgtggacga agtaccgaaa ggtcttaccg gaaaactcga
5161 cgcaagaaaa atcagagaga tcctcataaa ggccaagaag ggcggaaaga tcgccgtgga
5221 gcagctcgag agtataatca actttgaaaa actgactgaa tggaccagtt ctaatgttat
5281 ggaagagagg aagatcaaag tgtacttacc tcgcatgaag atggaggaaa aatacaacct
5341 cacatctgtc ttaatggcta tgggcattac tgacgtgttt agctcttcag ccaatctgtc
5401 tggcatctcc tcagcagaga gcctgaagat atctcaagct gtccatgcag cacatgcaga
5461 aatcaatgaa gcaggcagag aggtggtagg gtcagcagag gctggagtgg atgctgcaag
5521 cgtctctgaa gaatttaggg ctgaccatcc attcctcttc tgtatcaaac acatcgcaac
5581 caacgccgtt ctcttctttg gcagatgtgt tggatccgag cagctggagt ccatctacag
5641 gtactacggc ctgctgctga aggaaagatc cgaacaaaag cttatttctg aagaggactt
5701 gtaagcggcc gaaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
5761 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
5821 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
5881 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
5941 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
6001 ccacgcgccc tgtagcggcg cattaaataa cttcgtataa tgtatgctat acgaagttat
6061 atcgtatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct
6121 aactgacaca cattccacag aattccttga aatacatcca caggtacacc tccaattgac
6181 tcaaatgatg tcaattagtc tatcagaagc ttctaaagcc atgacatcat tttctggaat
6241 tttccaagct gtttaaaggc acagtcaact tagtgtatgt aaacttctga cccactggaa
6301 ttgtgataca gtgaattata agtgaaataa tctgtctgta aacaattgtt ggaaaaatga
6361 cttgtgtcat gcacaaagta gatgtcctaa ctgacttgcc aaaactattg tttgttaaca
6421 agaaatttgt ggagtagttg aaaaacgagt tttaatgact ccaacttaag tgtatgtaaa
6481 cttccgactt caactgtagg gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc
6541 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
6601 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
6661 tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
6721 accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
6781 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
6841 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
6901 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
6961 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
7021 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
7081 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
7141 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
7201 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
7261 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
7321 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
7381 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
7441 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
7501 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
7561 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
7621 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
7681 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
7741 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
7801 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
7861 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
7921 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
7981 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
8041 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
8101 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
8161 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
8221 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
8281 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
8341 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
8401 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
8461 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
8521 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
8581 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
8641 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
8701 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
8761 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
8821 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
8881 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
8941 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
9001 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
9061 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
9121 gaaacagcta tgaccatgat tacgccaagc ttgcatgcct gcaggtcgac tctagaccta
9181 tacagttgaa gtcggaagtt tacatacact taagttggag tcattaaaac tcgtttttca
9241 actactccac aaatttcttg ttaacaaaca atagttttgg caagtcagtt aggacatcta
9301 ctttgtgcat gacacaagtc atttttccaa caattgttta cagacagatt atttcactta
9361 taattcactg tatcacaatt ccagtgggtc agaagtttac atacactaag ttgactgtgc
9421 ctttaaacag cttggaaaat tccagaaaat gatgtcatgg ctttagaagc ttctgataga
9481 ctaattgaca tcatttgagt caattggagg tgtacctgtg gatgtatttc aaggggatcc
9541 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agcggccg
//
