| 质粒名称: | pNHEJ-RPG |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 85931 |
| 存储实验室: | Zhiying Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pXL-Bac II |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达；dual-reporter surrogate |
| 筛选标记: | 嘌呤霉素 |
| 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: | pPB-CMV-DsRed-polyA-CAG-NHEJ. Puro-T2A-EGFP,DsRed, CAG-NHEJ. Puro-T2A-EGFP |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | pCAG-F (5'-GCAACGTGCTGGTTATTGTG-3') |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pNHEJ-RPG was a gift from Zhiying Zhang (Addgene plasmid # 85931 ; http://n2t.net/addgene:85931 ; RRID:Addgene_85931) |
| 参考文献部分: | Dual-reporter surrogate systems for efficient enrichment of genetically modified cells. Ren C, Xu K, Liu Z, Shen J, Han F, Chen Z, Zhang Z. Cell Mol Life Sci. 2015 Jul;72(14):2763-72. doi: 10.1007/s00018-015-1874-6. Epub 2015 Mar 1. 10.1007/s00018-015-1874-6 PubMed 25725802 |
买家导航
pNHEJRPG载体基本信息
pNHEJRPG载体图谱质粒图谱和多克隆位点信息
pNHEJRPG载体简介
pNHEJRPG载体序列
LOCUS pNHEJ-RPG 9062 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Constitutively express DsRed and conditionally express Puro and
EGFP. Used to test transfection efficiency and enrich genetically
modified cells.
ACCESSION .
VERSION .
KEYWORDS pNHEJ-RPG
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9062)
AUTHORS Ren C, Xu K, Liu Z, Shen J, Han F, Chen Z, Zhang Z
TITLE Dual-reporter surrogate systems for efficient enrichment of
genetically modified cells.
JOURNAL Cell Mol Life Sci. 2015 Jul;72(14):2763-72. doi:
10.1007/s00018-015-1874-6. Epub 2015 Mar 1.
PUBMED 25725802
REFERENCE 2 (bases 1 to 9062)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9062
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 26..130
/gene="bla"
/label=AmpR promoter
CDS 131..991
/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(349..368)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1162..1750
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1651..1670
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1904..1921
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 2038..2059
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2074..2104
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2112..2128
/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 2117..2139
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2136..2152
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2136..2152
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 2173..2191
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
repeat_region 2340..2374
/label=piggyBac left (5') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
enhancer 2837..3140
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3141..3344
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 3295..3315
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 3378..3387
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3384..4064
/codon_start=1
/product="wild-type DsRed"
/label=DsRed1
/note="mammalian codon-optimized"
/translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
HLFL"
primer_bind complement(3567..3587)
/label=DsRed1-N
/note="DsRed1, reverse primer"
primer_bind 3785..3804
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 3979..4002
/label=DsRed1-C
/note="DsRed1, forward primer"
primer_bind complement(4068..4087)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 4133..4188
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(4187..4206)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
enhancer 4561..4940
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 4942..5219
/label=chicken beta-actin promoter
intron 5220..6236
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 6160..6182
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 6244..6263
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 6292..6301
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 6318..6911
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind 6811..6831
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
CDS 6936..6989
/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 6990..7706
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIDDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(7032..7053)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(7293..7312)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 7640..7661
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 7824..7945
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7861..7880)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 7915..7934
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
repeat_region 8295..8357
/label=piggyBac right (3') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
primer_bind complement(8423..8442)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(8424..8442)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(8449..8466)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(8449..8465)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8458..8480)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 8607..9062
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(8694..8713)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 8904..8925
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
61 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
361 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
661 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
1021 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
1081 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
1141 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
1201 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
1261 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
1321 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
1381 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
1441 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
1501 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
1561 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
1621 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
1681 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
1741 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
1801 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
1861 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
1921 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
1981 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
2041 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
2101 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
2161 cgccaagctc gaaattaacc ctcactaaag ggaacaaaag ctggtacctc gcgcgacttg
2221 gtttgccatt ctttagcgcg cgtcgcgtca cacagcttgg ccacaatgtg gtttttgtca
2281 aacgaagatt ctatgacgtg tttaaagttt aggtcgagta aagcgcaaat cttttttaac
2341 cctagaaaga tagtctgcgt aaaattgacg catgcattct tgaaatattg ctctctcttt
2401 ctaaatagcg cgaatccgtc gctgtgcatt taggacatct cagtcgccgc ttggagctcc
2461 cgtgaggcgt gcttgtcaat gcggtaagtg tcactgattt tgaactataa cgaccgcgtg
2521 agtcaaaatg acgcatgatt atcttttacg tgacttttaa gatttaactc atacgataat
2581 tatattgtta tttcatgttc tacttacgtg ataacttatt atatatatat tttcttgtta
2641 tagatatcgt gactaatata taataaaatg ggtagttctt tagacgatga gcatatcctc
2701 tctgctcttc tgcaaagcga tgacgagctt gttggctagc ggtggcggcc tcgacattga
2761 ttattgacta gagcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
2821 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
2881 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
2941 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
3001 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
3061 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
3121 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
3181 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
3241 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
3301 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
3361 agatccgcta gccggtcgcc accatggtgc gctcctccaa gaacgtcatc aaggagttca
3421 tgcgcttcaa ggtgcgcatg gagggcaccg tgaacggcca cgagttcgag atcgagggcg
3481 agggcgaggg ccgcccctac gagggccaca acaccgtgaa gctgaaggtg accaagggcg
3541 gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc tccaaggtgt
3601 acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc gagggcttca
3661 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
3721 ccctgcagga cggctgcttc atctacaagg tgaagttcat cggcgtgaac ttcccctccg
3781 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag cgcctgtacc
3841 cccgcgacgg cgtgctgaag ggcgagatcc acaaggccct gaagctgaag gacggcggcc
3901 actacctggt ggagttcaag tccatctaca tggccaagaa gcccgtgcag ctgcccggct
3961 actactacgt ggactccaag ctggacatca cctcccacaa cgaggactac accatcgtgg
4021 agcagtacga gcgcaccgag ggccgccacc acctgttcct gtaaggatct ttttccctct
4081 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
4141 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
4201 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
4261 tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg
4321 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
4381 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
4441 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
4501 ctcttatgga gatccctcga cctgcagccc aagcttatcg atactagcgg tggcggcctc
4561 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
4621 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
4681 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
4741 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
4801 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
4861 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
4921 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
4981 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
5041 tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg
5101 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
5161 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
5221 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
5281 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
5341 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
5401 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
5461 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
5521 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
5581 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
5641 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
5701 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
5761 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
5821 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
5881 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
5941 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
6001 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
6061 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
6121 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
6181 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
6241 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc accatggcgg
6301 ccgcgatatc ggatcccacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
6361 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
6421 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
6481 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
6541 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
6601 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
6661 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
6721 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
6781 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
6841 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
6901 agcccggtgc cgctcgagct accggtggca gtggagaggg cagaggaagt ctgctaacat
6961 gcggtgacgt cgaggagaat cctggcccag tgagcaaggg cgaggagctg ttcaccgggg
7021 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
7081 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
7141 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
7201 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
7261 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
7321 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
7381 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
7441 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
7501 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcgacgacg
7561 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
7621 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
7681 tcggcatgga cgagctgtac aagtaatcta gataactgat cataatcagc cataccacat
7741 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
7801 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
7861 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
7921 tgtccaaact catcaatgta tcttagtcga gttaattaac gagagcataa tattgatatg
7981 tgccaaagtt gtttctgact gactaataag tataatttgt ttctattatg tataggttaa
8041 gctaattact tattttataa tacaacatga ctgtttttaa agtacaaaat aagtttattt
8101 ttgtaaaaga gagaatgttt aaaagttttg ttactttata gaagaaattt tgagtttttg
8161 ttttttttta ataaataaat aaacataaat aaattgtttg ttgaatttat tattagtatg
8221 taagtgtaaa tataataaaa cttaatatct attcaaatta ataaataaac ctcgatatac
8281 agaccgataa aacacatgcg tcaattttac gcatgattat ctttaacgta cgtcacaata
8341 tgattatctt tctagggtta aataatagtt tctaattttt ttattattca gcctgctgtc
8401 gtgaataccg agctccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt
8461 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
8521 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
8581 gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
8641 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
8701 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
8761 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
8821 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
8881 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
8941 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
9001 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
9061 tt
//
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