| 質粒名稱: | XLone-BSD Cas13d P2A mCherry |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 155183 |
| 存儲實驗室: | Xiaoping Bao |
|
載體骨架基本信息 |
|
| 載體名稱: | pN1 |
| 載體出品公司: | Clontech |
| 空載體大小: | 6487 bp |
| 完整質粒大小: | 9385 |
| 篩選標記: | Blasticidin |
| 載體抗性: | 氨芐霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷貝數: | 高拷貝 |
|
插入基因信息 |
|
| 插入基因名稱: | CasRx、Cas13d |
| 物種來源: | H. sapiens (human) |
| 插入大小: | 2898bp |
| 啟動子: | TRE3G |
| 標簽/融合蛋白: |
NLS (C terminal on insert)
P2A mCherry (C terminal on insert)
|
| 克隆方法: | 限制性內切酶 |
| 5’克隆位點: | KpnI (not destroyed) |
| 3’克隆位點: | NheI, SpeI (not destroyed) |
| 5’測序引物 | gcgcctataaaagagtgctga |
| 3’測序引物 |
M13 FWD
(Common Sequencing Primers)
|
|
文獻引用方法 |
|
| 材料和方法部分: | XLone-BSD Cas13d P2A mCherry was a gift from Xiaoping Bao (Addgene plasmid # 155183 ; //n2t.net/addgene:155183 ; RRID:Addgene_155183) |
| 參考文獻部分: | Chemically-defined generation of human hemogenic endothelium and definitive hematopoietic progenitor cells. Chang Y, Syahirah R, Oprescu SN, Wang X, Jung J, Cooper SH, Torregrosa-Allen S, Elzey BD, Hsu AY, Randolph LN, Sun Y, Kuang S, Broxmeyer HE, Deng Q, Lian X, Bao X. Biomaterials. 2022 May 6;285:121569. doi: 10.1016/j.biomaterials.2022.121569. 10.1016/j.biomaterials.2022.121569 PubMed 35567999 |
買家導航
XLone-BSD Cas13d P2A mCherry載體質粒基本信息
XLone-BSD Cas13d P2A mCherry質粒圖譜載體圖譜和XLone-BSD Cas13d P2A mCherry載體序列質粒序列多克隆位點信息
XLone-BSD Cas13d P2A mCherry質粒載體簡介
XLone-BSD Cas13d P2A mCherry質粒序列載體序列
LOCUS XLone-BSD Cas13d P2A mCherry 9385 bp DNA circular SYN 14-APR-2025
DEFINITION PiggybacTransposon-based tunable and temporal expression control of
Cas13d and mCherry.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9385)
AUTHORS Chang Y, Syahirah R, Oprescu SN, Wang X, Jung J, Cooper SH,
Torregrosa-Allen S, Elzey BD, Hsu AY, Randolph LN, Sun Y, Kuang S,
Broxmeyer HE, Deng Q, Lian X, Bao X
TITLE Chemically-defined generation of human hemogenic endothelium and
definitive hematopoietic progenitor cells.
JOURNAL Biomaterials. 2022 May 6;285:121569. doi:
10.1016/j.biomaterials.2022.121569.
PUBMED 35567999
REFERENCE 2 (bases 1 to 9385)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
//gt12365.cn
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Biomaterials."; date: "2022-05-6"; volume: "285:121569. doi";
pages: " 10.1016/j.biomaterials.2022.121569"
COMMENT Sequence Label: XLone-BSD Cas13d P2A mCherry
FEATURES Location/Qualifiers
source 1..9385
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 9..376
/label=TRE3GS promoter
/note="3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On(R) 3G, modified to eliminate
binding sites for endogenous mammalian transcription
factors"
protein_bind 17..35
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 53..71
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 89..107
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 125..143
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 161..179
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 197..215
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 233..251
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
CDS 389..3286
/codon_start=1
/product="RNA-targeting Type VI CRISPR protein from
Ruminococcus flavefaciens strain XPD3002 (Konermann et al.,
2018)"
/label=RfxCas13d
/translation="MIEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDS
IRSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLE
KRYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFSTV
YTYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYII
NYGNECYDILALLSGLRHWVVHNNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITN
ELTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRK
DMSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFV
INLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPA
GRDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAF
FKDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDE
NGNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQ
KKQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGRE
NAEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLE
EKGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKK
AEEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFRNKAVHLEVARYVH
AYINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCV
PFGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGN"
CDS 3287..3334
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 3350..3406
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 3410..3433
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 3434..4141
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
GRHSTGGMDELYK"
primer_bind complement(3576..3594)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 3844..3863
/label=mCherry-F
/note="mCherry, forward primer"
polyA_signal 4347..4481
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
repeat_region 4757..4791
/label=piggyBac left (5') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
primer_bind complement(4802..4819)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4802..4818)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4811..4833)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(5027..5046)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5146..5168
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5206..5224)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5292..5396
/gene="bla"
/label=AmpR promoter
CDS join(5397..5465,5466..6257)
/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(5615..5634)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6428..7016
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6917..6936
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7170..7187
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 7304..7325
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter join(7340..7345,7346..7363,7364..7370)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7378..7394
/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 7383..7405
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 7402..7418
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7402..7418
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
repeat_region 7444..7506
/label=piggyBac right (3') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
polyA_signal 7682..7763
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
CDS complement(7880..8278)
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
CDS complement(8279..8335)
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS complement(8345..9088)
/codon_start=1
/product="modified rtTA protein that binds tightly to
promoters containing the tet operator in the presence of
doxycycline"
/label=Tet-On(R) 3G
/translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
DALDDFDLDMLPADALDDFDLDMLPG"
CDS complement(9089..9112)
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
regulatory 9112..9121
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter complement(9141..9352)
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
ORIGIN
1 atcacctcga gtttactccc tatcagtgat agagaacgta tgaagagttt actccctatc
61 agtgatagag aacgtatgca gactttactc cctatcagtg atagagaacg tataaggagt
121 ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa
181 cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc
241 agtgatagag aacgtataag ctttgcttat gtaaaccagg gcgcctataa aagagtgctg
301 attttttgag taaacttcaa ttccacaaca cttttgtctt ataccaactt tccgtaccac
361 ttcctaccct cgtaaaggta ccgccaccat gatcgaaaaa aaaaagtcct tcgccaaggg
421 catgggcgtg aagtccacac tcgtgtccgg ctccaaagtg tacatgacaa ccttcgccga
481 aggcagcgac gccaggctgg aaaagatcgt ggagggcgac agcatcagga gcgtgaatga
541 gggcgaggcc ttcagcgctg aaatggccga taaaaacgcc ggctataaga tcggcaacgc
601 caaattcagc catcctaagg gctacgccgt ggtggctaac aaccctctgt atacaggacc
661 cgtccagcag gatatgctcg gcctgaagga aactctggaa aagaggtact tcggcgagag
721 cgctgatggc aatgacaata tttgtatcca ggtgatccat aacatcctgg acattgaaaa
781 aatcctcgcc gaatacatta ccaacgccgc ctacgccgtc aacaatatct ccggcctgga
841 taaggacatt attggattcg gcaagttctc cacagtgtat acctacgacg aattcaaaga
901 ccccgagcac catagggccg ctttcaacaa taacgataag ctcatcaacg ccatcaaggc
961 ccagtatgac gagttcgaca acttcctcga taaccccaga ctcggctatt tcggccaggc
1021 ctttttcagc aaggagggca gaaattacat catcaattac ggcaacgaat gctatgacat
1081 tctggccctc ctgagcggac tgaggcactg ggtggtccat aacaacgaag aagagtccag
1141 gatctccagg acctggctct acaacctcga taagaacctc gacaacgaat acatctccac
1201 cctcaactac ctctacgaca ggatcaccaa tgagctgacc aactccttct ccaagaactc
1261 cgccgccaac gtgaactata ttgccgaaac tctgggaatc aaccctgccg aattcgccga
1321 acaatatttc agattcagca ttatgaaaga gcagaaaaac ctcggattca atatcaccaa
1381 gctcagggaa gtgatgctgg acaggaagga tatgtccgag atcaggaaaa atcataaggt
1441 gttcgactcc atcaggacca aggtctacac catgatggac tttgtgattt ataggtatta
1501 catcgaagag gatgccaagg tggctgccgc caataagtcc ctccccgata atgagaagtc
1561 cctgagcgag aaggatatct ttgtgattaa cctgaggggc tccttcaacg acgaccagaa
1621 ggatgccctc tactacgatg aagctaatag aatttggaga aagctcgaaa atatcatgca
1681 caacatcaag gaatttaggg gaaacaagac aagagagtat aagaagaagg acgcccctag
1741 actgcccaga atcctgcccg ctggccgtga tgtttccgcc ttcagcaaac tcatgtatgc
1801 cctgaccatg ttcctggatg gcaaggagat caacgacctc ctgaccaccc tgattaataa
1861 attcgataac atccagagct tcctgaaggt gatgcctctc atcggagtca acgctaagtt
1921 cgtggaggaa tacgcctttt tcaaagactc cgccaagatc gccgatgagc tgaggctgat
1981 caagtccttc gctagaatgg gagaacctat tgccgatgcc aggagggcca tgtatatcga
2041 cgccatccgt attttaggaa ccaacctgtc ctatgatgag ctcaaggccc tcgccgacac
2101 cttttccctg gacgagaacg gaaacaagct caagaaaggc aagcacggca tgagaaattt
2161 cattattaat aacgtgatca gcaataaaag gttccactac ctgatcagat acggtgatcc
2221 tgcccacctc catgagatcg ccaaaaacga ggccgtggtg aagttcgtgc tcggcaggat
2281 cgctgacatc cagaaaaaac agggccagaa cggcaagaac cagatcgaca ggtactacga
2341 aacttgtatc ggaaaggata agggcaagag cgtgagcgaa aaggtggacg ctctcacaaa
2401 gatcatcacc ggaatgaact acgaccaatt cgacaagaaa aggagcgtca ttgaggacac
2461 cggcagggaa aacgccgaga gggagaagtt taaaaagatc atcagcctgt acctcaccgt
2521 gatctaccac atcctcaaga atattgtcaa tatcaacgcc aggtacgtca tcggattcca
2581 ttgcgtcgag cgtgatgctc aactgtacaa ggagaaaggc tacgacatca atctcaagaa
2641 actggaagag aagggattca gctccgtcac caagctctgc gctggcattg atgaaactgc
2701 ccccgataag agaaaggacg tggaaaagga gatggctgaa agagccaagg agagcattga
2761 cagcctcgag agcgccaacc ccaagctgta tgccaattac atcaaataca gcgacgagaa
2821 gaaagccgag gagttcacca ggcagattaa cagggagaag gccaaaaccg ccctgaacgc
2881 ctacctgagg aacaccaagt ggaatgtgat catcagggag gacctcctga gaattgacaa
2941 caagacatgt accctgttca gaaacaaggc cgtccacctg gaagtggcca ggtatgtcca
3001 cgcctatatc aacgacattg ccgaggtcaa ttcctacttc caactgtacc attacatcat
3061 gcagagaatt atcatgaatg agaggtacga gaaaagcagc ggaaaggtgt ccgagtactt
3121 cgacgctgtg aatgacgaga agaagtacaa cgataggctc ctgaaactgc tgtgtgtgcc
3181 tttcggctac tgtatcccca ggtttaagaa cctgagcatc gaggccctgt tcgataggaa
3241 cgaggccgcc aagttcgaca aggagaaaaa gaaggtgtcc ggcaataagc gacctgccgc
3301 cacaaagaag gctggacagg ctaagaagaa gaaagctagc ggcagcggcg ccactaactt
3361 ctccctgttg aaacaagcag gggatgtcga agagaatccc gggccaatgg actacaagga
3421 cgacgatgac aaggtgagca agggcgagga ggataacatg gccatcatca aggagttcat
3481 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
3541 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
3601 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta
3661 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa
3721 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
3781 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga
3841 cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc
3901 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca
3961 ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
4021 ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga
4081 acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta
4141 agaattccga tcatattcaa taacccacta gtagaccacc tcccctgcga gctaagctgg
4201 acagccaatg acgggtaaga gagtgacatt tttcactaac ctaagacagg agggccgtca
4261 gagctactgc ctaatccaaa gacgggtaaa agtgataaaa atgtatcact ccaacctaag
4321 acaggcgcag cttccgaggg atttgagatc cagacatgat aagatacatt gatgagtttg
4381 gacaaaccaa aactagaatg cagtgaaaaa aatgccttat ttgtgaaatt tgtgatgcta
4441 ttgccttatt tgtaaccatt ataagctgca ataaacaagt ttgatatcta taacaagaaa
4501 atatatatat aataagttat cacgtaagta gaacatgaaa taacaatata attatcgtat
4561 gagttaaatc ttaaaagtca cgtaaaagat aatcatgcgt cattttgact cacgcggtcg
4621 ttatagttca aaatcagtga cacttaccgc attgacaagc acgcctcacg ggagctccaa
4681 gcggcgactg agatgtccta aatgcacagc gacggattcg cgctatttag aaagagagag
4741 caatatttca agaatgcatg cgtcaatttt acgcagacta tctttctagg gttaagaatt
4801 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
4861 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
4921 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
4981 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
5041 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
5101 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
5161 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
5221 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
5281 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
5341 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
5401 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
5461 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
5521 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
5581 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
5641 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
5701 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
5761 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
5821 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
5881 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
5941 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
6001 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
6061 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
6121 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
6181 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
6241 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
6301 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
6361 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
6421 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
6481 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
6541 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
6601 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
6661 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
6721 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
6781 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
6841 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
6901 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
6961 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
7021 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
7081 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
7141 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
7201 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
7261 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
7321 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
7381 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aaggtcgact
7441 taaccctaga aagataatca tattgtgacg tacgttaaag ataatcatgc gtaaaattga
7501 cgcatgtgtt ttatcggtct gtatatcgag gtttatttat taatttgaat agatattaag
7561 ttttattata tttacactta catactaata ataaattcaa caaacaattt atttatgttt
7621 atttatttat taaaaaaaaa caaaaactca aaatttcttc tataaagtaa caaaactttt
7681 agcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
7741 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
7801 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtatggctg
7861 attatgatcc tctggagatt tagccctccc acacataacc agagggcagc aattcacgaa
7921 tcccaactgc cgtcggctgt ccatcactgt ccttcactat ggctttgatc ccaggatgca
7981 gatcgagaag cacctgtcgg caccgtccgc aggggctcaa gatgcccctg ttctcatttc
8041 cgatcgcgac gatacaagtc aggttgccag ctgccgcagc agcagcagtg cccagcacca
8101 cgagttctgc acaaggtccc ccagtaaaat gatatacatt gacaccagtg aagatgcggc
8161 cgtcgctaga gagagctgcg ctggcgacgc tgtagtcttc agagatgggg atgctgttga
8221 ttgtagccgt tgctctttca atgagggtgg attcttcttg agacaaaggc ttggccatgg
8281 ggccggggtt ctcctccacg tcgccggcct gcttcagcag gctgaagttg gtggcgccgc
8341 tgcccccggg gagcatgtca aggtcaaaat cgtcaagagc gtcagcaggc agcatatcaa
8401 ggtcaaagtc gtcaagggca tcggctggga gcatgtctaa gtcaaaatcg tcaagggcgt
8461 cggtcggccc gccgctttcg cactttagct gtttctccag gccacatatg attagttcca
8521 ggccgaaaag gaaggcaggt tcggctccct gccggtcgaa cagctcaatt gcttgtttca
8581 gaagtggggg catagaatcg gtggtaggtg tctctctttc ctcttttgct acttgatgct
8641 cctgttcctc caatacgcag cccagtgtaa agtggcccac ggcggacaga gcgtacagtg
8701 cgttctccag ggagaagcct tgctgacaca ggaacgcgag ctgattttcc agggtttcgt
8761 actgtttctc tgttgggcgg gtgccgagat gcactttagc cccgtcgcga tgtgagagga
8821 gagcacagcg gtatgacttg gcgttgttcc gcagaaagtc ttgccatgac tcgccttcca
8881 gggggcagga gtgggtatga tgcctgtcca gcatctcgat tggcagggca tcgagcaggg
8941 cccgcttgtt cttcacgtgc cagtacaggg taggctgctc aactcccagc ttttgagcga
9001 gtttccttgt cgtcaggcct tcgataccga ctccattgag taattccaga gcagagttta
9061 tgactttgct cttgtccagt ctagacatct tatcgtcatc gtctttgtaa tccatggtgg
9121 cggatcccgc gtcacgacac ctgtgttctg gcggcaaacc cgttgcgaaa aagaacgttc
9181 acggcgacta ctgcacttat atacggttct cccccaccct cgggaaaaag gcggagccag
9241 tacacgacat cactttccca gtttaccccg cgccaccttc tctaggcacc ggttcaattg
9301 ccgacccctc cccccaactt ctcggggact gtgggcgatg tgcgctctgc ccactgacgg
9361 gcaccggagc cactcgagtg gaatt
//