| 质粒名称: | p0521s |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 62862 |
| 存储实验室: | Hamilton Smith |
|
载体骨架基本信息 |
|
| 载体名称: | pCC1BAC |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | 16310 bp |
| 载体修饰: | 添加酵母Cen6-ArsH4-His3，P.tricornutum R1R2，从RP4转移的来源，P.tricornutum ShBle盒 |
| 载体类型: | 合成生物学；三角褐指藻的外显体载体 |
| 筛选标记: | Zeocin, URA3, HIS3 |
| 载体抗性: | 氯霉素，25μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | 用于诱导型高拷贝表达的Epi300 |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
|
文献引用方法 |
|
| 材料和方法部分: | p0521s was a gift from Hamilton Smith (Addgene plasmid # 62862 ; http://n2t.net/addgene:62862 ; RRID:Addgene_62862) |
| 参考文献部分: | Designer diatom episomes delivered by bacterial conjugation. Karas BJ, Diner RE, Lefebvre SC, McQuaid J, Phillips AP, Noddings CM, Brunson JK, Valas RE, Deerinck TJ, Jablanovic J, Gillard JT, Beeri K, Ellisman MH, Glass JI, Hutchison CA 3rd, Smith HO, Venter JC, Allen AE, Dupont CL, Weyman PD. Nat Commun. 2015 Apr 21;6:6925. doi: 10.1038/ncomms7925. 10.1038/ncomms7925 PubMed 25897682 |
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 p0521s 16310 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Episome vector for Phaeodactylum tricornutum (pCC1BAC-based).
ACCESSION .
VERSION .
KEYWORDS p0521s
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 16310)
AUTHORS Karas BJ, Diner RE, Lefebvre SC, McQuaid J, Phillips AP, Noddings
CM, Brunson JK, Valas RE, Deerinck TJ, Jablanovic J, Gillard JT,
Beeri K, Ellisman MH, Glass JI, Hutchison CA 3rd, Smith HO, Venter
JC, Allen AE, Dupont CL, Weyman PD
TITLE Designer diatom episomes delivered by bacterial conjugation.
JOURNAL Nat Commun. 2015 Apr 21;6:6925. doi: 10.1038/ncomms7925.
PUBMED 25897682
REFERENCE 2 (bases 1 to 16310)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..16310
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 6..620
/label=oriV
/note="origin of replication for the bacterial F plasmid"
rep_origin 696..915
/label=ori2
/note="secondary origin of replication for the bacterial F
plasmid; also known as oriS"
CDS 1006..1761
/codon_start=1
/gene="repE"
/product="replication initiation protein for the bacterial
F plasmid"
/label=repE
/translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
misc_feature 1764..2014
/gene="incC"
/label=incC
/note="incompatibility region of the bacterial F plasmid"
misc_feature 2176..2679
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 2680..2866
/gene="S. cerevisiae HIS3"
/label=HIS3 promoter
CDS 2867..3529
/codon_start=1
/gene="S. cerevisiae HIS3"
/product="imidazoleglycerol-phosphate dehydratase, required
for histidine biosynthesis"
/label=HIS3
/note="yeast auxotrophic marker"
/translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
CDS 3719..4894
/codon_start=1
/gene="sopA"
/product="partitioning protein for the bacterial F plasmid"
/label=sopA
/translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
CDS 4894..5865
/codon_start=1
/gene="sopB"
/product="partitioning protein for the bacterial F plasmid"
/label=sopB
/translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
misc_feature 5938..6411
/gene="sopC"
/label=sopC
/note="centromere-like partitioning region of the bacterial
F plasmid"
misc_feature 6671..7069
/label=cos
/note="lambda cos site; allows packaging into phage lambda
particles"
protein_bind 7087..7120
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
primer_bind 7188..7207
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
CDS complement(7287..7562)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSYLGETIEYSSLHACRSTLEDPRVPSSNSPYSESYYNSL
AVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS"
primer_bind 7400..7422
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 7414..7431
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 7415..7431
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7438..7457
/label=T7
/note="T7 promoter, forward primer"
promoter 7438..7456
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 7459..7515
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(7558..7574)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7558..7574)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7571..7593)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7582..7598
/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(7606..7636)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7651..7672
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(7892..8551)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 8466..8485
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(8552..8654)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
promoter 9530..9548
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
CDS 10252..10626
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
promoter 11354..11569
/gene="S. cerevisiae URA3"
/label=URA3 promoter
CDS 11570..12373
/codon_start=1
/gene="S. cerevisiae URA3"
/product="orotidine-5'-phosphate decarboxylase, required
for uracil biosynthesis"
/label=URA3
/note="yeast auxotrophic marker, counterselectable with
5-fluoroorotic acid (5-FOA)"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
oriT 13131..13240
/note="incP origin of transfer"
CDS 13273..13644
/codon_start=1
/gene="traJ"
/product="oriT-recognizing protein"
/label=traJ
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
ORIGIN
1 tcgagctggt tgccctcgcc gctgggctgg cggccgtcta tggccctgca aacgcgccag
61 aaacgccgtc gaagccgtgt gcgagacacc gcggccggcc gccggcgttg tggatacctc
121 gcggaaaact tggccctcac tgacagatga ggggcggacg ttgacacttg aggggccgac
181 tcacccggcg cggcgttgac agatgagggg caggctcgat ttcggccggc gacgtggagc
241 tggccagcct cgcaaatcgg cgaaaacgcc tgattttacg cgagtttccc acagatgatg
301 tggacaagcc tggggataag tgccctgcgg tattgacact tgaggggcgc gactactgac
361 agatgagggg cgcgatcctt gacacttgag gggcagagtg ctgacagatg aggggcgcac
421 ctattgacat ttgaggggct gtccacaggc agaaaatcca gcatttgcaa gggtttccgc
481 ccgtttttcg gccaccgcta acctgtcttt taacctgctt ttaaaccaat atttataaac
541 cttgttttta accagggctg cgccctgtgc gcgtgaccgc gcacgccgaa ggggggtgcc
601 cccccttctc gaaccctccc ggtcgagtga gcgaggaagc accagggaac agcacttata
661 tattctgctt acacacgatg cctgaaaaaa cttcccttgg ggttatccac ttatccacgg
721 ggatattttt ataattattt tttttatagt ttttagatct tcttttttag agcgccttgt
781 aggcctttat ccatgctggt tctagagaag gtgttgtgac aaattgccct ttcagtgtga
841 caaatcaccc tcaaatgaca gtcctgtctg tgacaaattg cccttaaccc tgtgacaaat
901 tgccctcaga agaagctgtt ttttcacaaa gttatccctg cttattgact cttttttatt
961 tagtgtgaca atctaaaaac ttgtcacact tcacatggat ctgtcatggc ggaaacagcg
1021 gttatcaatc acaagaaacg taaaaatagc ccgcgaatcg tccagtcaaa cgacctcact
1081 gaggcggcat atagtctctc ccgggatcaa aaacgtatgc tgtatctgtt cgttgaccag
1141 atcagaaaat ctgatggcac cctacaggaa catgacggta tctgcgagat ccatgttgct
1201 aaatatgctg aaatattcgg attgacctct gcggaagcca gtaaggatat acggcaggca
1261 ttgaagagtt tcgcggggaa ggaagtggtt ttttatcgcc ctgaagagga tgccggcgat
1321 gaaaaaggct atgaatcttt tccttggttt atcaaacgtg cgcacagtcc atccagaggg
1381 ctttacagtg tacatatcaa cccatatctc attcccttct ttatcgggtt acagaaccgg
1441 tttacgcagt ttcggcttag tgaaacaaaa gaaatcacca atccgtatgc catgcgttta
1501 tacgaatccc tgtgtcagta tcgtaagccg gatggctcag gcatcgtctc tctgaaaatc
1561 gactggatca tagagcgtta ccagctgcct caaagttacc agcgtatgcc tgacttccgc
1621 cgccgcttcc tgcaggtctg tgttaatgag atcaacagca gaactccaat gcgcctctca
1681 tacattgaga aaaagaaagg ccgccagacg actcatatcg tattttcctt ccgcgatatc
1741 acttccatga cgacaggata gtctgagggt tatctgtcac agatttgagg gtggttcgtc
1801 acatttgttc tgacctactg agggtaattt gtcacagttt tgctgtttcc ttcagcctgc
1861 atggattttc tcatactttt tgaactgtaa tttttaagga agccaaattt gagggcagtt
1921 tgtcacagtt gatttccttc tctttccctt cgtcatgtga cctgatatcg ggggttagtt
1981 cgtcatcatt gatgagggtt gattatcaca gtttattact ctgaattggc tatccgcgtg
2041 tgtacctcta cctggagttt ttcccacggt ggatatttct tcttgcgctg agcgtaagag
2101 ctatctgaca gaacagttct tctttgcttc ctcgccagtt cgctcgctat gctcggttac
2161 acggctgcgg cgagcatcac gtgctataaa aataattata atttaaattt tttaatataa
2221 atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt
2281 ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta ccttgctctt
2341 cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag accacacacg
2401 aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta tttaatctgc
2461 ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt tttaaacctt
2521 tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac tttctaaaat
2581 ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta ttccatcatt
2641 aaaagatacg aggcgcgtgt aagttacagg caagcgatcc tagtacactc tatatttttt
2701 tatgcctcgg taatgatttt catttttttt ttccacctag cggatgactc tttttttttc
2761 ttagcgattg gcattatcac ataatgaatt atacattata taaagtaatg tgatttcttc
2821 gaagaatata ctaaaaaatg agcaggcaag ataaacgaag gcaaagatga cagagcagaa
2881 agccctagta aagcgtatta caaatgaaac caagattcag attgcgatct ctttaaaggg
2941 tggtccccta gcgatagagc actcgatctt cccagaaaaa gaggcagaag cagtagcaga
3001 acaggccaca caatcgcaag tgattaacgt ccacacaggt atagggtttc tggaccatat
3061 gatacatgct ctggccaagc attccggctg gtcgctaatc gttgagtgca ttggtgactt
3121 acacatagac gaccatcaca ccactgaaga ctgcgggatt gctctcggtc aagcttttaa
3181 agaggcccta ggggccgtgc gtggagtaaa aaggtttgga tcaggatttg cgcctttgga
3241 tgaggcactt tccagagcgg tggtagatct ttcgaacagg ccgtacgcag ttgtcgaact
3301 tggtttgcaa agggagaaag taggagatct ctcttgcgag atgatcccgc attttcttga
3361 aagctttgca gaggctagca gaattaccct ccacgttgat tgtctgcgag gcaagaatga
3421 tcatcaccgt agtgagagtg cgttcaaggc tcttgcggtt gccataagag aagccacctc
3481 gcccaatggt accaacgatg ttccctccac caaaggtgtt cttatgtagt tttacacagg
3541 agtctggact tgacgctagt gataataagt gactgaggta tgtgctcttc ttatctcctt
3601 ttgtagtgtt gctcttattt taaacaactt tgcggttttt tgatgacttt gcgattttgt
3661 tgttgctttg cagtaaattg caagatttaa taaaaaaacg caaagcaatg attaaaggat
3721 gttcagaatg aaactcatgg aaacacttaa ccagtgcata aacgctggtc atgaaatgac
3781 gaaggctatc gccattgcac agtttaatga tgacagcccg gaagcgagga aaataacccg
3841 gcgctggaga ataggtgaag cagcggattt agttggggtt tcttctcagg ctatcagaga
3901 tgccgagaaa gcagggcgac taccgcaccc ggatatggaa attcgaggac gggttgagca
3961 acgtgttggt tatacaattg aacaaattaa tcatatgcgt gatgtgtttg gtacgcgatt
4021 gcgacgtgct gaagacgtat ttccaccggt gatcggggtt gctgcccata aaggtggcgt
4081 ttacaaaacc tcagtttctg ttcatcttgc tcaggatctg gctctgaagg ggctacgtgt
4141 tttgctcgtg gaaggtaacg acccccaggg aacagcctca atgtatcacg gatgggtacc
4201 agatcttcat attcatgcag aagacactct cctgcctttc tatcttgggg aaaaggacga
4261 tgtcacttat gcaataaagc ccacttgctg gccggggctt gacattattc cttcctgtct
4321 ggctctgcac cgtattgaaa ctgagttaat gggcaaattt gatgaaggta aactgcccac
4381 cgatccacac ctgatgctcc gactggccat tgaaactgtt gctcatgact atgatgtcat
4441 agttattgac agcgcgccta acctgggtat cggcacgatt aatgtcgtat gtgctgctga
4501 tgtgctgatt gttcccacgc ctgctgagtt gtttgactac acctccgcac tgcagttttt
4561 cgatatgctt cgtgatctgc tcaagaacgt tgatcttaaa gggttcgagc ctgatgtacg
4621 tattttgctt accaaataca gcaatagcaa tggctctcag tccccgtgga tggaggagca
4681 aattcgggat gcctggggaa gcatggttct aaaaaatgtt gtacgtgaaa cggatgaagt
4741 tggtaaaggt cagatccgga tgagaactgt ttttgaacag gccattgatc aacgctcttc
4801 aactggtgcc tggagaaatg ctctttctat ttgggaacct gtctgcaatg aaattttcga
4861 tcgtctgatt aaaccacgct gggagattag ataatgaagc gtgcgcctgt tattccaaaa
4921 catacgctca atactcaacc ggttgaagat acttcgttat cgacaccagc tgccccgatg
4981 gtggattcgt taattgcgcg cgtaggagta atggctcgcg gtaatgccat tactttgcct
5041 gtatgtggtc gggatgtgaa gtttactctt gaagtgctcc ggggtgatag tgttgagaag
5101 acctctcggg tatggtcagg taatgaacgt gaccaggagc tgcttactga ggacgcactg
5161 gatgatctca tcccttcttt tctactgact ggtcaacaga caccggcgtt cggtcgaaga
5221 gtatctggtg tcatagaaat tgccgatggg agtcgccgtc gtaaagctgc tgcacttacc
5281 gaaagtgatt atcgtgttct ggttggcgag ctggatgatg agcagatggc tgcattatcc
5341 agattgggta acgattatcg cccaacaagt gcttatgaac gtggtcagcg ttatgcaagc
5401 cgattgcaga atgaatttgc tggaaatatt tctgcgctgg ctgatgcgga aaatatttca
5461 cgtaagatta ttacccgctg tatcaacacc gccaaattgc ctaaatcagt tgttgctctt
5521 ttttctcacc ccggtgaact atctgcccgg tcaggtgatg cacttcaaaa agcctttaca
5581 gataaagagg aattacttaa gcagcaggca tctaaccttc atgagcagaa aaaagctggg
5641 gtgatatttg aagctgaaga agttatcact cttttaactt ctgtgcttaa aacgtcatct
5701 gcatcaagaa ctagtttaag ctcacgacat cagtttgctc ctggagcgac agtattgtat
5761 aagggcgata aaatggtgct taacctggac aggtctcgtg ttccaactga gtgtatagag
5821 aaaattgagg ccattcttaa ggaacttgaa aagccagcac cctgatgcga cctcgtttta
5881 gtctacgttt atctgtcttt acttaatgtc ctttgttaca ggccagaaag cataactggc
5941 ctgaatattc tctctgggcc cactgttcca cttgtatcgt cggtctgata atcagactgg
6001 gaccacggtc ccactcgtat cgtcggtctg attattagtc tgggaccacg gtcccactcg
6061 tatcgtcggt ctgattatta gtctgggacc acggtcccac tcgtatcgtc ggtctgataa
6121 tcagactggg accacggtcc cactcgtatc gtcggtctga ttattagtct gggaccatgg
6181 tcccactcgt atcgtcggtc tgattattag tctgggacca cggtcccact cgtatcgtcg
6241 gtctgattat tagtctggaa ccacggtccc actcgtatcg tcggtctgat tattagtctg
6301 ggaccacggt cccactcgta tcgtcggtct gattattagt ctgggaccac gatcccactc
6361 gtgttgtcgg tctgattatc ggtctgggac cacggtccca cttgtattgt cgatcagact
6421 atcagcgtga gactacgatt ccatcaatgc ctgtcaaggg caagtattga catgtcgtcg
6481 taacctgtag aacggagtaa cctcggtgtg cggttgtatg cctgctgtgg attgctgctg
6541 tgtcctgctt atccacaaca ttttgcgcac ggttatgtgg acaaaatacc tggttaccca
6601 ggccgtgccg gcacgttaac cgggctgcat ccgatgcaag tgtgtcgctg tcgacgagct
6661 cgcgagctcg gacatgaggt tgccccgtat tcagtgtcgc tgatttgtat tgtctgaagt
6721 tgtttttacg ttaagttgat gcagatcaat taatacgata cctgcgtcat aattgattat
6781 ttgacgtggt ttgatggcct ccacgcacgt tgtgatatgt agatgataat cattatcact
6841 ttacgggtcc tttccggtga tccgacaggt tacggggcgg cgacctcgcg ggttttcgct
6901 atttatgaaa attttccggt ttaaggcgtt tccgttcttc ttcgtcataa cttaatgttt
6961 ttatttaaaa taccctctga aaagaaagga aacgacaggt gctgaaagcg agctttttgg
7021 cctctgtcgt ttcctttctc tgtttttgtc cgtggaatga acaatggaag tccgagctca
7081 tcgctaataa cttcgtatag catacattat acgaagttat attcgatgcg gccgcaaggg
7141 gttcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
7201 gattgtactg agagtgcacc atatgcggtg tgaaatacca cacagatgcg taaggagaaa
7261 ataccgcatc aggcgccatt cgccattcag ctgcgcaact gttgggaagg gcgatcggtg
7321 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
7381 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa
7441 tacgactcac tatagggcga attcgagctc ggtacccggg gatcctctag agtcgacctg
7501 caggcatgca agcttgagta ttctatagtc tcacctaaat agcttggcgt aatcatggtc
7561 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
7621 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
7681 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
7741 ccaacgcgaa ccccttgcgg ccgcccgggc cgtcgaccaa ttctcatgtt tgacagctta
7801 tcatcgaatt tctgccattc atccgcttat tatcacttat tcaggcgtag caaccaggcg
7861 tttaagggca ccaataactg ccttaaaaaa attacgcccc gccctgccac tcatcgcagt
7921 actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa
7981 cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga
8041 aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca
8101 cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca
8161 ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt
8221 cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac
8281 aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg
8341 gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat
8401 ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac
8461 attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa
8521 cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg
8581 ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc
8641 ttacgtgccg atcaacgtct cattttcgcc aaaagttggc ccagggcttc ccggtatcaa
8701 cagggacacc aggatttatt tattctgcga agtgatcttc cgtcacaggt atttattcgc
8761 gataagctca tggagcggcg taaccgtcgc acaggaagga cagagaaagc gcggatctgg
8821 gaagtgacgg acagaacggt caggacctgg attggggagg cggttgccgc cgctgctgct
8881 gacggtgtga cgttctctgt tccggtcaca ccacatacgt tccgccattc ctatgcgatg
8941 cacatgctgt atgccggtat accgctgaaa gttctgcaaa gcctgatggg acataagtcc
9001 atcagttcaa cggaagtcta cacgaaggtt tttgcgctgg atgtggctgc ccggcaccgg
9061 gtgcagtttg cgatgccgga gtctgatgcg gttgcgatgc tgaaacaatt atcctgagaa
9121 taaatgcctt ggcctttata tggaaatgtg gaactgagtg gatatgctgt ttttgtctgt
9181 taaacagaga agctggctgt tatccactga gaagcgaacg aaacagtcgg gaaaatctcc
9241 cattatcgta gagatccgca ttattaatct caggagcctg tgtagcgttt ataggaagta
9301 gtgttctgtc atgatgcctg caagcggtaa cgaaaacgat ttgaatatgc cttcaggaac
9361 aatagaaatc ttcgtgcggt gttacgttga agtggagcgg attatgtcag caatggacag
9421 aacaacctaa tgaacacaga accatgatgt ggtctgtcct tttacagcca gtagtgctcg
9481 ccgcagtcga gcgacagggc gaagcccacc atgattacgc caagctcgaa attaaccctc
9541 actaaaggga acaaaagctg gtacctaaca ggattagtgc aattcgagtt gaatcactgg
9601 gaaaaacatt gtcttctttt ttatattatc atttgcatta gtgctgcagt cgtagatact
9661 tgttggttga aagacatcag ctgggaggga ctggactagc gtttggtaag gagacatacc
9721 tgttaacgtt ggttgcaaaa ttccatttcg cgatttatgt tatctgtaaa tcctgatttg
9781 tctggaattc ttgatacttc cgttttttta gaggccaatg attagcatcg gcgattctca
9841 aaatagcatt ttcgacatgc ggtgctgatt tcataaacat agacaacgct tttacatgta
9901 aaagtaactt gcggacttgg aacagtgctc tgtttttggt gtgaacgtaa ctcagcaata
9961 tttctgtgct agcaaggttt tttatgatcg accgaagatc tcaaaactcc gggtctttca
10021 actgtctgac tagaccatgt tcgtaacgtc ggacagcagc tttcgttgta ctggtagaat
10081 ttctacgtgc gaagcacgtg taggcaggtt gaacgacgat ccctgccgat ggatggattg
10141 gcacgcggcg gaacgctttc gtgatctaca ccacctggat cttcacatat cttcgaaatc
10201 gaaaaattaa ccaagtcgac ggtatcgata atattctagc tgagggtacc catggccaag
10261 ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg
10321 accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg
10381 gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg
10441 gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc
10501 acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg
10561 cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag
10621 gactgaccga cgccgaccaa caccgccggt ccgacgcggc ccgacgggtc cgaggcctcg
10681 gagatctggg cccatgcggc cgcaacaact acctcgactt tggctgggac actttcagtg
10741 aggacaagaa gcttcagaag cgtgctatcg aactcaacca gggacgtgcg gcacaaatgg
10801 gcatccttgc tctcatggtg cacgaacagt tgggagtctc tatccttcct taaaaattta
10861 attttcatta gttgcagtca ctccgctttg gtttcacagt caggaataac actagctcgt
10921 cttcacgtaa ctataacggt cctaaggtag cgaacgttgc aggccatgct gtccaggcag
10981 gtagatgacg accatcaggg acagcttcaa ggatcgctcg cggctcttac cagcctaact
11041 tcgatcattg gaccgctgat cgtcacggcg atttatgccg cctcggcgag cacatggaac
11101 gggttggcat ggattgtagg cgccgcccta taccttgtct gcctccccgc gttgcgtcgc
11161 ggtgcatgga gccgggccac ctcgacctga atggaagccg gcggcacctc gctaacggat
11221 tcaccactcc aagaattgga gccaatcaat tcttgcggag aactgtgaat gcgcaaacca
11281 acccttggca gaacatatcc atcgcgtccg ccatctccag cagccgcacg cggcgcatcg
11341 gggggggggg ggtttcaatt catcattttt tttttattct tttttttgat ttcggtttcc
11401 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
11461 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
11521 cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc
11581 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
11641 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
11701 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
11761 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
11821 caatttttta ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca
11881 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
11941 ggtgggccca ggtattgtta gcggtttgaa gcaggcggcg gaagaagtaa caaaggaacc
12001 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctagcta ctggagaata
12061 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
12121 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
12181 gggtttagat gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt
12241 ctctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
12301 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
12361 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
12421 agcttcaatt taattatatc agttattacc cggccgggaa tctcggtcgt aatgattttt
12481 ataatgacga aaaaaaaaaa aattggaaag aaaacccccc cccccccgca gcgttgggtc
12541 ctggccacgg gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg
12601 ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg
12661 ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta
12721 aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg
12781 atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc
12841 ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg
12901 ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt
12961 catcgagtta cgctagggat aacaggtaat atagcatctt ccgctgcata accctgcttc
13021 ggggtcatta tagcgatttt ttcggtatat ccatcctttt tcgcacgata tacaggattt
13081 tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc gggcaggata
13141 ggtgaagtag gcccacccgc gagcgggtgt tccttcttca ctgtccctta ttcgcacctg
13201 gcggtgctca acgggaatcc tgctctgcga ggctggccgg ctaccgccgg cgtaacagat
13261 gagggcaagc ggatggctga tgaaaccaag ccaaccagga agggcagccc acctatcaag
13321 gtgtactgcc ttccagacga acgaagagcg attgaggaaa aggcggcggc ggccggcatg
13381 agcctgtcgg cctacctgct ggccgtcggc cagggctaca aaatcacggg cgtcgtggac
13441 tatgagcacg tccgcgagct ggcccgcatc aatggcgacc tgggccgcct gggcggcctg
13501 ctgaaactct ggctcaccga cgacccgcgc acggcgcggt tcggtgatgc cacgatcctc
13561 gccctgctgg cgaagatcga agagaagcag gacgagcttg gcaaggtcat gatgggcgtg
13621 gtccgcccga gggcagagcc atgacttttt tagccgctaa aacggccggg gggtgcgcgt
13681 gattgccaag cacgtcccca tgcgctccat caagaagagg cacttcgagc tgtaagtaca
13741 tcaccgacga gcaaggcaag acgatccgcg cctgttttat tgagaacgtt gttcgtgttg
13801 gcctcaatgg tagcgatgcg tcattcagcg aagttctggt gctgatgatg tggttcgctt
13861 tgccactggt caatgtggta agcccgtgta atgtcagtaa cctttttact gatctcagct
13921 tgagcacggt cgctgatgag cttatccatg gccccacggt aacggatatg atcctctagg
13981 gcgttgacaa attctttgtc ggttttcatg gggtagacgt cagtgaccag ggaacgtctc
14041 cctacaaaaa gttgcgcata ctttgctccg ttgtcgacgg caggggtgtc ggaccagatt
14101 gtatctgtgg caacggcctc attgcgtcaa tggaccttta aagccgggaa acgagacttg
14161 aaatgtttac gtagaggtgc attatagacc tcacgcgcat attgagtggt tgcaaaaatg
14221 gttttgcgaa cggtatcact ggagacccat gccaggcaag gacggagggc atcataatca
14281 tgttcattgc gttggacagc atgcttgtta cacgtaagta tacggtcgag agcatcgtga
14341 cgttgagaat tacagatggc aacgtgtcgg tgcatgtatt ggccgagatt gtcaaatcgt
14401 ggctcaatat atggcatgcc aggtaagtca tggatgtcgg tatgccattc agccttcatg
14461 tcgatttcgt tgtcgagaac tgatgggtcc cagtcaacgt cagaagtaag aacaacatgg
14521 ggaagggaat gaagttcatg actggtgggt gcgcgcatat ccatgtacgg aagaccctgt
14581 cgtatgttta gtgggatgat gtaatcatcc agggtaacaa tacgttggtt gccgcccacg
14641 gtacgagacc gatcttgaac acaattgtgg caatgttcaa gttgggcgct ggaatggatg
14701 gttttacctt taccaagatg cgcgtattga tgcatgataa caacaatggg ccctcgttgc
14761 gattcgacaa gaccggcagc cgtgacaatg tctagattgg acaaggtatg ttcatttata
14821 ccggttatgt tggctgaacg tccagttttg tgcagtacgg taacatcact ccccgcaagt
14881 ccaccgttgg caccgcgatc gacaagtgca gatgtggtat tttgtactgt gtgtcgggag
14941 acttggtact gtaacacatt tgactgaagt gacttggtgg taccatgagg acggagggaa
15001 cccgttgata cggtgttgcg tgacgctgca agtaccttac gtatgtcgcc tggatccata
15061 cgtccaaccc gatcagaaag atgtgctagg agttccgttt ctggtgcaca atcatgaaag
15121 gtatccacgg gagttccatt gttgtcgtcc ggggctgcgt cgcgactgcc agttgcatgt
15181 acatgcgcca tcaagggacg tgccaatcca ttactaaccg ggagaacctt agctggggca
15241 gccaatccct gaagtatagc cttagcatca tctgagagct ggttccacat atctctaggg
15301 atataaggtc gttcacggtt agtgggcgta ctatgatgtc ggttattagt acaattctcc
15361 ctgcgggcat gcgcattggc ttcatagagt acggaaggtg acaagtcaat attgtagtta
15421 acatccggat cagtgtcaaa gtctgtagga tggtaagaaa gatcagtaga atgaatacta
15481 cgcttttcct tgggactacg ggaattagag aagttgtttc ctttattgta gagtgacgct
15541 gaagcaagta gaagactaag gtaactctcg tagctaatag gattacctcc tttggctagg
15601 tcaagagtgg ctgtgatctt cacttgacaa agttccggta cattgtggac agcattctcc
15661 aaaagactaa gacacagttg ctttgggagt tgctcagcca ttggtacagt attgtggtag
15721 atacaaaggt ggttcttcca atgaaggata aatccttctg ctgtgcctgt ccatgaggat
15781 ccatatttcg ccgtagttag gtaaccaagc gtggtggctg aactgatctt cgcacttgct
15841 gattccgtat agtgtttgac aactttacaa aacacttctt gcgcagttcg ctctagcctg
15901 gacagagaag ggacaatctt cgtcgttaag actcgtgcga atagcaaaag atcacaaaat
15961 agcacatcgg caccgaccaa cgattatttc caaggaaaaa aagaatgctt cactacaaga
16021 aattgtgtca tccctataca gagtcttgtt actgtgacag aaaattgatg gaagatgtgg
16081 cggattgcct ttacactagc caacttgttc gactaattgc agcttcttct gagaggcttc
16141 accgagtaac gcgaagaaca ccggtgtctc gtacatgctc gtcggtgaac gctcgtccaa
16201 tgacaccccc cactttgtat caatatccca acttggtagt gaactggaat gatacatgca
16261 atttcgcgcc gcatcaacag ccacgggcca ccatcgacga atagactcgg
//
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