| 质粒名称: | pCW-eGFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 162823 |
| 存储实验室: | Matthew Steinhauser |
|
载体骨架基本信息 |
|
| 载体名称: | Addgene plasmid pCW-Cas9 (plasmid#50661) |
| 载体出品公司: | Clontech |
| 空载体大小: | 7600bp |
| 完整质粒大小: | 8329bp |
| 载体类型: | 哺乳细胞表达、 Lentiviral |
| 筛选标记: | Puromycin |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 30 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | eGFP |
| 插入大小: | 720 bp |
| 基因突变: | V163A |
| 启动子: | Tet ON |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NheI(未破坏) |
| 3’克隆位点: | BamHI(未破坏) |
| 5’测序引物 | pCW Seq F: AGCTCGTTTAGTGAACCGTCAGATC |
| 3’测序引物 | pCW Seq R: GAACGGACGTGAAGAATGTG |
|
文献引用方法 |
|
| 材料和方法部分: | pCW-eGFP was a gift from Matthew Steinhauser (Addgene plasmid # 162823 ; http://n2t.net/addgene:162823 ; RRID:Addgene_162823) |
| 参考文献部分: | Prolonged fasting drives a program of metabolic inflammation in human adipose tissue. Fazeli PK, Zhang Y, O'Keefe J, Pesaresi T, Lun M, Lawney B, Steinhauser ML. Mol Metab. 2020 Sep 28;42:101082. doi: 10.1016/j.molmet.2020.101082. 10.1016/j.molmet.2020.101082 PubMed 32992039 |
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 pCW-eGFP 8329 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Doxycycline-inducible overexpression of eGFP.
ACCESSION .
VERSION .
KEYWORDS pCW-eGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8329)
AUTHORS Fazeli PK, Zhang Y, O'Keefe J, Pesaresi T, Lun M, Lawney B,
Steinhauser ML
TITLE Prolonged fasting drives a program of metabolic inflammation in
human adipose tissue.
JOURNAL Mol Metab. 2020 Sep 28;42:101082. doi: 10.1016/j.molmet.2020.101082.
PUBMED 32992039
REFERENCE 2 (bases 1 to 8329)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8329
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 1..118
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 172..486
/label=tight TRE promoter
/note="Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter"
protein_bind 180..198
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 216..234
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 251..269
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 287..305
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 323..341
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 358..376
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 394..412
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 459..483
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 505..1224
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(550..571)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(811..830)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1158..1179
/label=EGFP-C
/note="EGFP, forward primer"
promoter 1231..1741
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 1660..1678
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 1751..2347
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(1751..1770)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 2247..2267
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
CDS 2348..2401
/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 2402..3148
/codon_start=1
/product="improved tetracycline-controlled transactivator"
/label=rtTA-Advanced
/note="In the Tet-On(R) system, rtTA-Advanced binds to the
Tet-responsive element and stimulates transcription only in
the presence of tetracycline or doxycycline."
/translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
DALDDFDLDMLPADALDDFDLDMLPG"
primer_bind complement(3188..3208)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(3606..3617)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 3822..4055
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4127..4248
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4164..4183)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4218..4237
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 4288..4423
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4350..4369
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 4423..4494
/gene="bla"
/label=AmpR promoter
CDS 4495..5355
/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(4713..4732)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5526..6114
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6015..6034
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6268..6285
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 6402..6423
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6438..6468
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6476..6492
/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 6481..6503
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 6500..6516
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6500..6516
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 6535..6555
/label=T3
/note="T3 promoter, forward primer"
promoter 6537..6555
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 6583..6809
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 6810..6990
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 7037..7162
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 7655..7888
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 8073..8117
/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"
ORIGIN
1 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
61 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
121 ggtttattac agggacagca gagatccact ttggccgcga atcgatatgt cgagtttact
181 ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgatg
241 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
301 agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttatcc
361 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
421 cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt gaaccgtcag
481 atcgcctgga gaattggcta gcccatggtg agcaagggcg aggagctgtt caccggggtg
541 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtctggc
601 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
661 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
721 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
781 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
841 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
901 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
961 atcatggccg acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacatc
1021 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
1081 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
1141 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
1201 ggcatggacg agctgtacaa gtaaggatcc ggggttgggg ttgcgccttt tccaaggcag
1261 ccctgggttt gcgcagggac gcggctgctc tgggcgtggt tccgggaaac gcagcggcgc
1321 cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc gtcacccgga tcttcgccgc
1381 tacccttgtg ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg aaggttcctt
1441 gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta gtaccctcgc
1501 agacggacag cgccagggag caatggcagc gcgccgaccg cgatgggctg tggccaatag
1561 cggctgctca gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc gggaggcggg
1621 gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat tctgcaagcc
1681 tccggagcgc acgtcggcag tcggctccct cgttgaccga atcaccgacc tctctcccca
1741 gcaattcacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
1801 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
1861 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
1921 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
1981 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
2041 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
2101 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
2161 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
2221 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
2281 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
2341 cggtgccgaa ggtagaggtt ctctcctcac ttgtggtgat gttgaagaaa accctggtcc
2401 aatgtctaga ctggacaaga gcaaagtcat aaacggagct ctggaattac tcaatggtgt
2461 cggtatcgaa ggcctgacga caaggaaact cgctcaaaag ctgggagttg agcagcctac
2521 cctgtactgg cacgtgaaga acaagcgggc cctgctcgat gccctgccaa tcgagatgct
2581 ggacaggcat catacccact tctgccccct ggaaggcgag tcatggcaag actttctgcg
2641 gaacaacgcc aagtcatacc gctgtgctct cctctcacat cgcgacgggg ctaaagtgca
2701 tctcggcacc cgcccaacag agaaacagta cgaaaccctg gaaaatcagc tcgcgttcct
2761 gtgtcagcaa ggcttctccc tggagaacgc actgtacgct ctgtccgccg tgggccactt
2821 tacactgggc tgcgtattgg aggaacagga gcatcaagta gcaaaagagg aaagagagac
2881 acctaccacc gattctatgc ccccacttct gagacaagca attgagctgt tcgaccggca
2941 gggagccgaa cctgccttcc ttttcggcct ggaactaatc atatgtggcc tggagaaaca
3001 gctaaagtgc gaaagcggcg ggccgaccga cgcccttgac gattttgact tagacatgct
3061 cccagccgat gcccttgacg actttgacct tgatatgctg cctgctgacg ctcttgacga
3121 ttttgacctt gacatgctcc ccgggtaact aagtaaggat cgatccaaga tatcgtattc
3181 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
3241 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
3301 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
3361 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
3421 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
3481 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
3541 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
3601 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
3661 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
3721 cgcctgtttc gcctcggcgt ccggactaga ggtaccttta agaccaatga cttacaaggc
3781 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
3841 ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg
3901 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
3961 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
4021 cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat
4081 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
4141 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
4201 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
4261 ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
4321 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
4381 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctttcaaa tatgtatccg
4441 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
4501 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
4561 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
4621 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
4681 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
4741 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
4801 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
4861 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
4921 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
4981 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
5041 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
5101 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
5161 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
5221 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
5281 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
5341 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
5401 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
5461 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
5521 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
5581 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
5641 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
5701 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
5761 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
5821 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
5881 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
5941 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
6001 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
6061 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
6121 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
6181 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
6241 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
6301 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
6361 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
6421 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
6481 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
6541 aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac
6601 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
6661 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
6721 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
6781 tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct
6841 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
6901 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
6961 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
7021 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
7081 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
7141 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
7201 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
7261 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
7321 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
7381 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
7441 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
7501 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
7561 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
7621 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
7681 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
7741 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
7801 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
7861 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
7921 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
7981 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
8041 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
8101 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
8161 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
8221 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
8281 cgaggggaca attctcgacc tcgagacaaa tggcagtatt catccacaa
//
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