| 质粒名称: | AAVS1-Puro cTnT FUCCI |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 136935 |
| 存储实验室: | Xiaoping Bao |
|
载体骨架基本信息 |
|
| 载体名称: | AAVS1-Puro |
| 载体出品公司: | -- |
| 空载体大小: | 11266 bp |
| 完整质粒大小: | 11878 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，合成生物学 |
| 筛选标记: | 嘌呤霉素 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | Troponin T promoter,cTnT |
| 物种来源: | H. sapiens (human), Synthetic |
| 插入大小: | 612 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | cTnT |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | SalI(未破坏) |
| 3’克隆位点: | MluI (未破坏) |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | AAVS1-Puro cTnT FUCCI was a gift from Xiaoping Bao (Addgene plasmid # 136935 ; http://n2t.net/addgene:136935 ; RRID:Addgene_136935) |
| 参考文献部分: | Fluorescent indicators for continuous and lineage-specific reporting of cell-cycle phases in human pluripotent stem cells. Chang Y, Hellwarth PB, Randolph LN, Sun Y, Xing Y, Zhu W, Lian XL, Bao X. Biotechnol Bioeng. 2020 Apr 11. doi: 10.1002/bit.27352. 10.1002/bit.27352 PubMed 32277708 |
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 AAVS1-Puro cTnT FUCCI 11876 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION donor plasmid for cardiac-specific FUCCI expression in human cells.
ACCESSION .
VERSION .
KEYWORDS AAVS1-Puro cTnT FUCCI
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11876)
AUTHORS Chang Y, Hellwarth PB, Randolph LN, Sun Y, Xing Y, Zhu W, Lian XL,
Bao X
TITLE Fluorescent indicators for continuous and lineage-specific reporting
of cell-cycle phases in human pluripotent stem cells.
JOURNAL Biotechnol Bioeng. 2020 Apr 11. doi: 10.1002/bit.27352.
PUBMED 32277708
REFERENCE 2 (bases 1 to 11876)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..11876
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 1..837
/label=HA-R
/note="right homology arm from the adeno-associated virus
integration site (AAVS1) within intron 1 of the human
PPP1R12C gene"
protein_bind 911..932
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 947..977
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 985..1001
/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 1009..1025
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 1368..1387
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
polyA_signal 1386..1441
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind 1487..1506
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 1616..2092
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind 1874..1893
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
misc_feature 2124..2712
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS 2230..2241
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind 2639..2659
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2750..3022)
/codon_start=1
/product="degron consisting of residues 30-120 of human
Cdt1 (Zielke and Edgar, 2015)"
/label=Cdt1 (30-120)
/translation="PSPARPALRAPASATSGSRKRARPPAAPGRDQARPPARRRLRLSV
DEVSSPSTPEAPDIPACPSPGQKIKKSTPAAGQPPHLTSAQDQDTI"
CDS complement(3077..3727)
/codon_start=1
/product="monomeric Kusabira-Orange 2 fluorescent protein"
/label=mKO2
/translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAH"
misc_feature 3742..4314
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(3901..3920)
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
primer_bind 4130..4147
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
CDS complement(4319..4648)
/codon_start=1
/product="degron consisting of residues 1-110 of human
Geminin (Zielke and Edgar, 2015)"
/label=Gem1 (1-110)
/translation="MNPSMKQKQEEIKENIKNSSVPRRTLKMIQPSASGSLVGRENELS
AGLSKRKHRNDHLTSTTSSPGVIVPESSENKNLGGVTQESFDLMIKENPSSQYWKEVAE
KRRKAL"
CDS complement(4679..5395)
/codon_start=1
/product="bright green-yellow fluorescent protein derived
from GFP (Lam et al., 2012)"
/label=Clover
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
EFVTAAGITHGMDELYK"
primer_bind complement(4721..4742)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 5329..5350
/label=EGFP-N
/note="EGFP, reverse primer"
protein_bind complement(5411..5435)
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
promoter complement(5467..5485)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 6129..6353
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 6342..6359
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
CDS complement(6391..6990)
/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(6474..6494)
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
primer_bind 6971..6990
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
CDS complement(7000..7053)
/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"
misc_feature 7077..7102
/label=SA
/note="splice acceptor site"
misc_feature 7109..7912
/label=HA-L
/note="left homology arm from the adeno-associated virus
integration site (AAVS1) within intron 1 of the human
PPP1R12C gene"
promoter complement(7951..7969)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(7990..8006)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 8014..8030
/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(8038..8068)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8083..8104
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8221..8238)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8392..8980)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8472..8491)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS 9104..9964
/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(9322..9341)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
primer_bind complement(10325..10344)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 10939..10958
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
CDS complement(11361..11663)
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind complement(11463..11482)
/label=ccdB-fwd
/note="ccdB gene, forward primer"
primer_bind 11786..11808
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 11800..11817
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 11801..11817
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 11824..11842
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 agagcagagc caggaacccc tgtagggaag gggcaggaga gccaggggca tgagatggtg
61 gacgaggaag ggggacaggg aagcctgagc gcctctcctg ggcttgccaa ggactcaaac
121 ccagaagccc agagcagggc cttagggaag cgggaccctg ctctgggcgg aggaatatgt
181 cccagatagc actggggact ctttaaggaa agaaggatgg agaaagagaa agggagtaga
241 ggcggccacg acctggtgaa cacctaggac gcaccattct cacaaaggga gttttccaca
301 cggacacccc cctcctcacc acagccctgc caggacgggg ctggctactg gccttatctc
361 acaggtaaaa ctgacgcacg gaggaacaat ataaattggg gactagaaag gtgaagagcc
421 aaagttagaa ctcaggacca acttattctg attttgtttt tccaaactgc ttctcctctt
481 gggaagtgta aggaagctgc agcaccagga tcagtgaaac gcaccagaca gccgcgtcag
541 agcagctcag gttctgggag agggtagcgc agggtggcca ctgagaaccg ggcaggtcac
601 gcatcccccc cttccctccc accccctgcc aagctctccc tcccaggatc ctctctggct
661 ccatcgtaag caaaccttag aggttctggc aaggagagag atggctccag gaaatggggg
721 tgtgtcacca gataaggaat ctgcctaaca ggaggtgggg gttagaccca atatcaggag
781 actaggaagg aggaggccta aggatggggc ttttctgtca ccaatcctgt ccctagtaaa
841 gcttagtact gtcgatccgc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
901 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
961 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
1021 atgaccatga ttacgccaag cttgggctgc aggtcgaggg atctccataa gagaagaggg
1081 acagctatga ctgggagtag tcaggagagg aggaaaaatc tggctagtaa aacatgtaag
1141 gaaaatttta gggatgttaa agaaaaaaat aacacaaaac aaaatataaa aaaaatctaa
1201 cctcaagtca aggcttttct atggaataag gaatggacag cagggggctg tttcatatac
1261 tgatgacctc tttatagcca acctttgttc atggcagcca gcatatgggc atatgttgcc
1321 aaactctaaa ccaaatactc attctgatgt tttaaatgat ttgccctccc atatgtcctt
1381 ccgagtgaga gacacaaaaa attccaacac actattgcaa tgaaaataaa tttcctttat
1441 tagccagaag tcagatgctc aaggggcttc atgatgtccc cataattttt ggcagaggga
1501 aaaagatctc agtggtattt gtgagccagg gcattggcca caccagccac caccttctga
1561 taggcagcct gcacctgagg agtgaattca cgcgcctggt tacctacaaa atcagaagga
1621 cagggaaggg agcagtggtt cacgcctgta atcccagcaa tttgggaggc caaggtgggt
1681 agatcacctg agattaggag ttggagacca gcctggccaa tatggtgaaa ccccgtctct
1741 accaaaaaaa caaaaattag ctgagcctgg tcatgcatgc ctggaatccc aacaactcgg
1801 gaggctgagg caggagaatc gcttgaaccc aggaggcgga gattgcagtg agccaagatt
1861 gtgccactgc actccagctt ggttcccaat agaccccgca ggccctacag gttgtcttcc
1921 caacttgccc cttgctccat accacccccc tccaccccat aatattatag aaggacacct
1981 agtcagacaa aatgatgcaa cttaatttta ttaggacaag gctggtgggc actggagtgg
2041 caacttccag ggccaggaga ggcactgggg aggggtcaca gggatgccac ccgtagatct
2101 ctcgagcagc gctcggtatc gatgcgggga ggcggcccaa agggagatcc gactcgtctg
2161 agggcgaagg cgaagacgcg gaagaggccg cagagccggc agcaggccgc gggaaggaag
2221 gtccgctgga ttgagggccg aagggacgta gcagaaggac gtcccgcgca gaatccaggt
2281 ggcaacatag gcgagcagcc aaggaaagga cgatgatttc cccgacaaca ccacggaatt
2341 gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg cgatgagttc
2401 cgccgtggca atagggaggg ggaaagcgaa agtcccggaa aggagctgac aggtggtggc
2461 aatgccccaa ccagtggggg ttgcgtcagc aaacacagtg cacaccacgc cacgttgcct
2521 gacaacgggc cacaactcct cataaagaga cagcaaccag gatttataca aggaggagaa
2581 aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag cgtatccaca
2641 tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa ttttgtaatc
2701 cagaggttga ttatcgataa gcttgatatc gaattcacgc gtaaactcag atggtgtcct
2761 ggtcctgcgc ggatgtcagg tggggcggct gacctgctgc cggggtggat ttctttatct
2821 tctggcccgg agaagggcag gctgggatgt ctggggcctc gggggtactg gggctggaaa
2881 cctcgtccac cgacagccgc agtctcctgc gggccggtgg cctggcctgg tcgcgtccgg
2941 gggcggcggg cgggcgggcg cgcttgcggc tgccactggt agcggaggcc ggggcgcgga
3001 gtgcgggcct ggcggggctg ggctcgagcg gccgccagtg tgcatggata tcgcccaggg
3061 aaggcaacat tgagtagtgg gccacggcgt cctccacctg ctcggtgatg ttgccctcgg
3121 tcttcctcac cagcctgtgg ccgatgtagt ggtcgccggg catctccagg atctccttgg
3181 cggccttgta ggtggtcttc atctggcact tgtggttgcc gccgccctcc agcttcaggt
3241 acatggtcac gtcgcccttc agcacgccgt cgctggcggt gatcttctcg gtggagggct
3301 cccagtccac gctctggttc tgcatgatgg ggccgtcggc ggggaagttc acgccggtga
3361 acttggactt gtggtagaag gtgttgcccc tcaggctgat gtgggcgctc acggaggcgg
3421 agccgccgtc ctcgaactcc agggacctct cccaggacag gccctcagga aaggcctgct
3481 tgaaatagtc tgggatctct tctgggtact tggtaaaaac tctgtggccg taacagaaca
3541 cgtgggacac caggtcgaag gcgaaaggca ttggcccgcc ctcggccatt gtgacgcgca
3601 gtgtcatctc ctgatgtccc tcgtaaggtc tgcctgtgcc ctcaccctcg attgtgaact
3661 catgcccatt gacggagccg tccatgtagt acctcatctt catctcgggc ttgatcacgc
3721 tcaccatgaa ttctgtggcc attatcatcg tgtttttcaa aggaaaacca cgtccccgtg
3781 gttcgggggg cctagacgtt ttttaacctc gactaaacac atgtaaagca tgtgcaccga
3841 ggccccagat cagatcccat acaatggggt accttctggg catccttcag ccccttgttg
3901 aatacgcttg aggagagcca tttgactctt tccacaacta tccaactcac aacgtggcac
3961 tggggttgtg ccgcctttgc aggtgtatct tatacacgtg gcttttggcc gcagaggcac
4021 ctgtcgccag gtggggggtt ccgctgcctg caaagggtcg ctacagacgt tgtttgtctt
4081 caagaagctt ccagaggaac tgcttccttc acgacattca acagaccttg cattcctttg
4141 gcgagagggg aaagacccct aggaatgctc gtcaagaaga cagggccagg tttccgggcc
4201 ctcacattgc caaaagacgg caatatggtg gaaaataaca tatagacaaa cgcacaccgg
4261 ccttattcca agcggcttcg gccagtaacg ttaggggggg gggagggaga ggggcttaca
4321 gcgcctttct ccgtttttct gccacttcct tccaatactg agaggatgga ttttctttaa
4381 tcataagatc aaatgactcc tgggtgactc ctccaagatt tttattttca ctagattctg
4441 ggacaataac cccagggctg gaagttgtag atgttaagtg gtcattccga tgtttccttt
4501 tggacaagcc tgcggacagc tcattttctc ttccaacaag agatccagat gcagaaggct
4561 gaatcatctt cagagttctt cttgggacag aactattctt tatattctct ttgatttctt
4621 cttgtttctg cttcatactg ggattcatct cgagcggccg ccagtgtgca tggatatcct
4681 tgtacagctc gtccatgcca tgtgtaatcc cggcggcggt cacgaactcc agcaggacca
4741 tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctgatggctc aggtagtggt
4801 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
4861 gctgcacgct gccgtcctca acgttgtggc ggatcttgaa gttagccttg atgccgttct
4921 tctgcttgtc ggccgtgata tagacgttgt ggctgttgaa gttgtactcc agcttgtgcc
4981 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
5041 tgtcgccctc gaacttcacc tcggcgcggg tcttgtaggt accgtcgtcc ttgaaagaga
5101 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
5161 tgtggtcggg gtagcggctg aagcaggcca cgccgtagcc gaaggtggtc acgagggtgg
5221 gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgttgg
5281 tggcatcgcc ctcgccctcg ccgcggacgc tgaacttgtg gccgtttacg tcgccgtcca
5341 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgc
5401 tagcgtcgac agcctgcttt tttgtacaaa cttgttgatc aagcttacct agccagcttg
5461 ggtctcccta tagtgagtcg tattactcga gggtggcgga tcctggaggc gtctgctcag
5521 tctcagcggg gactgggtga ggcagaggat ggagagggct ttaagcaggc atgtgggctg
5581 gggcctggtg agccagccct gcggagggag gaatgtgcga caggggacgg gtggggcagg
5641 gggatggcgg tgggggtggg gggtgttggc tgctattttg gcaggtgcca gggacaaggc
5701 tacaggaaca tgtaccccac gccatataag cccatgtggt cctccagctg ctcagataag
5761 ctatttaaaa ccagagcaga tatgcaggga acagtcatgc aacataaacc agctgtccct
5821 cttgagaatc ctgataaagc agaggccagc aacccaggcc tgggagggcc agctgggagc
5881 agggttgggg ggcagaaggc aacctccaag acactccata agtctcagca ccagaatctt
5941 ggaaggcaga gggcaagagt tatgtgctgc tccacttgaa ctgatgctgg gggtaaagac
6001 atcttccagg ctactggctc ctaatggact gagcagcctt gggcaggttg ccggctctgc
6061 cagccccagt gaggacatct gcaaggtggg tcttctccat gacactagtc aataatcaat
6121 gtcgagaccc atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc
6181 ccccttgctg tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg
6241 cgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa
6301 ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaggc
6361 tgatcagcgg gtttaaacgg gccctctaga tcaggcaccg ggcttgcggg tcatgcacca
6421 ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta
6481 gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg cgcgctcggc
6541 cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga
6601 gacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc
6661 ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc gcgggccgat
6721 ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc
6781 gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc
6841 ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg acggtgtggc gcgtggcggg
6901 gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt cgcgggtggc
6961 gaggcgcacc gtgggcttgt actcggtcat ctcgagccta gggccgggat tctcctccac
7021 gtcaccgcat gttagaagac ttcctctgcc ctctccgctg ccagatctct cgaggccctg
7081 tgggaggaag agaagaggtc agaagcttgc cccactgtgg ggtggagggg acagataaaa
7141 gtacccagaa ccagagccac attaaccggc cctgggaata taaggtggtc ccagctcggg
7201 gacacaggat ccctggaggc agcaaacatg ctgtcctgaa gtggacatag gggcccgggt
7261 tggaggaaga agactagctg agctctcgga cccctggaag atgccatgac agggggctgg
7321 aagagctagc acagactaga gaggtaaggg gggtagggga gctgcccaaa tgaaaggagt
7381 gagaggtgac ccgaatccac aggagaacgg ggtgtccagg caaagaaagc aagaggatgg
7441 agaggtggct aaagccaggg agacggggta ctttggggtt gtccagaaaa acggtgatga
7501 tgcaggccta caagaagggg aggcgggacg caagggagac atccgtcgga gaaggccatc
7561 ctaagaaacg agagatggca caggccccag aaggagaagg aaaagggaac ccagcgagtg
7621 aagacggcat ggggttgggt gagggaggag agatgcccgg agaggaccca gacacgggga
7681 ggatccgctc agaggacatc acgtggtgca gcgccgagaa ggaagtgctc cggaaagagc
7741 atccttgggc agcaacacag cagagagcaa ggggaagagg gagtggagga agacggaacc
7801 tgaaggaggc ggcagggaag gatctgggcc agccgtagag gtgacccagg ccacaagctg
7861 cagacagaaa gcggcacagg cccaggggag agaatgctgg tcagagaaag caaagggcga
7921 attcgtttaa acctgcagga ctagctagtc cctttagtga gggttaattc tgagcttggc
7981 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
8041 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
8101 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
8161 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
8221 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
8281 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
8341 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
8401 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
8461 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
8521 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
8581 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
8641 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
8701 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
8761 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
8821 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
8881 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
8941 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
9001 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagaca
9061 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
9121 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
9181 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
9241 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
9301 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
9361 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt
9421 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
9481 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
9541 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
9601 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
9661 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
9721 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
9781 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
9841 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
9901 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
9961 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
10021 atttaaaagg atctaggtga agatcctttt tgataatctc atgagcggat acatatttga
10081 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
10141 tgatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt
10201 aagcgttaat aattcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat
10261 cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt
10321 cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc
10381 cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat
10441 cgccatgcat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg
10501 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
10561 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
10621 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
10681 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
10741 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
10801 gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg
10861 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
10921 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
10981 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga
11041 tcagagcttg atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact
11101 ttgcagggct tcccaacctt accagagggc gccccagctg gcaattccgg ttcgcttgct
11161 gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc tacctgcttt
11221 ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt catccggggt
11281 cagcaccgtt tctgcggact ggctttctac gtgaaaagga tctaggtgaa gatccttttt
11341 gataatctca tgcctgacat ttatattccc cagaacatca ggttaatggc gtttttgatg
11401 tcattttcgc ggtggctgag atcagccact tcttccccga taacggagac cggcacactg
11461 gccatatcgg tggtcatcat gcgccagctt tcatccccga tatgcaccac cgggtaaagt
11521 tcacgggaga ctttatctga cagcagacgt gcactggcca gggggatcac catccgtcgc
11581 cccggcgtgt caataatatc actctgtaca tccacaaaca gacgataacg gctctctctt
11641 ttataggtgt aaaccttaaa ctgccgtacg tataggctgc gcaactgttg ggaagggcga
11701 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
11761 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
11821 ttgtaatacg actcactata gggcgaattg aatttagcgg ccgcgaattc gccctt
//
