| 质粒名称: | pCpf1b-sp |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 122188 |
| 存储实验室: | Cheng-Cai Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | 未知 |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 14674bp |
| 载体类型: | 细菌表达, CRISPR |
| 载体抗性: | Spectinomycin, 50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 克隆方法: | Ligation Independent Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | pCpf1b-sp was a gift from Cheng-Cai Zhang (Addgene plasmid # 122188 ; http://n2t.net/addgene:122188 ; RRID:Addgene_122188) |
| 参考文献部分: | Expanding the potential of CRISPR-Cpf1 based genome editing technology in the cyanobacterium Anabaena PCC 7120. Niu TC, Lin GM, Xie LR, Wang ZQ, Xing WY, Zhang JY, Zhang CC. ACS Synth Biol. 2018 Dec 10. doi: 10.1021/acssynbio.8b00437. 10.1021/acssynbio.8b00437 PubMed 30525474 |
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pCpf1b-sp载体质粒基本信息
pCpf1b-sp质粒图谱载体图谱和pCpf1b-sp载体序列质粒序列多克隆位点信息

pCpf1b-sp质粒载体简介
pCpf1b-sp质粒序列载体序列
LOCUS pCpf1b-sp 14674 bp DNA circular SYN 25-AUG-2024
DEFINITION Bacterial genome editing plasmid with spectinomycin resistance
marker and using sacB as a counter-selection marker.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14674)
AUTHORS Niu TC, Lin GM, Xie LR, Wang ZQ, Xing WY, Zhang JY, Zhang CC
TITLE Expanding the potential of CRISPR-Cpf1 based genome editing
technology in the cyanobacterium Anabaena PCC 7120.
JOURNAL ACS Synth Biol. 2018 Dec 10. doi: 10.1021/acssynbio.8b00437.
PUBMED 30525474
REFERENCE 2 (bases 1 to 14674)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
Biol. 2018 Dec 10. doi: 10.1021/acssynbio.8b00437."
FEATURES Location/Qualifiers
source 1..14674
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..446
/gene="sacR"
/label=sacB promoter
/note="sacB promoter and control region"
CDS 447..1868
/codon_start=1
/gene="Bacillus subtilis sacB"
/product="secreted levansucrase that renders bacterial
growth sensitive to sucrose"
/label=SacB
/note="negative selection marker"
/translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
VKDSILEQGQLTVNK"
primer_bind 2359..2378
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter 2482..2511
/label=lac promoter
/note="promoter for the E. coli lac operon"
promoter 6435..6469
/label=J23119 promoter
/note="bacterial promoter (Registry of Standard Biological
Parts BBa_J23119)"
CDS complement(6537..6833)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSG"
primer_bind 6683..6705
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 6697..6714
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 6698..6714
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6724..6743
/label=T7
/note="T7 promoter, forward primer"
promoter 6724..6742
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6790..6810)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(6790..6808)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(6829..6845)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6829..6845)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6842..6864)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6853..6869
/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(6877..6907)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6922..6943
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS 7653..8444
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 9223..9617
/label=RSF1010 oriV
/note="replication origin of the broad-host-range plasmid
RSF1010; requires the RSF1010 RepA/B/C proteins for
replication (Scholz et al., 1989)"
oriT 9959..10046
/label=RSF1010 oriT
/note="origin of transfer of the broad-host-range plasmid
RSF1010 (Scholz et al., 1989)"
CDS 11285..12256
/codon_start=1
/gene="repB"
/product="replication protein B of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
/label=RSF1010 RepB
/translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
CDS 12767..13606
/codon_start=1
/gene="repA"
/product="replication protein A of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
/label=RSF1010 RepA
/translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
CDS 13593..14444
/codon_start=1
/gene="repC"
/product="replication protein C of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
/label=RSF1010 RepC
/translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
AG"
ORIGIN
1 cacatatacc tgccgttcac tattatttag tgaaatgaga tattatgata ttttctgaat
61 tgtgattaaa aaggcaactt tatgcccatg caacagaaac tataaaaaat acagagaatg
121 aaaagaaaca gatagatttt ttagttcttt aggcccgtag tctgcaaatc cttttatgat
181 tttctatcaa acaaaagagg aaaatagacc agttgcaatc caaacgagag tctaatagaa
241 tgaggtcgaa aagtaaatcg cgcgggtttg ttactgataa agcaggcaag acctaaaatg
301 tgtaaagggc aaagtgtata ctttggcgtc accccttaca tattttaggt ctttttttat
361 tgtgcgtaac taacttgcca tcttcaaaca ggagggctgg aagaagcaga ccgctaacac
421 agtacataaa aaaggagaca tgaacgatga acatcaaaaa gtttgcaaaa caagcaacag
481 tattaacctt tactaccgca ctgctggcag gaggcgcaac tcaagcgttt gcgaaagaaa
541 cgaaccaaaa gccatataag gaaacatacg gcatttccca tattacacgc catgatatgc
601 tgcaaatccc tgaacagcaa aaaaatgaaa aatatcaagt tcctgaattc gattcgtcca
661 caattaaaaa tatctcttct gcaaaaggcc tggacgtttg ggacagctgg ccattacaaa
721 acgctgacgg cactgtcgca aactatcacg gctaccacat cgtctttgca ttagccggag
781 atcctaaaaa tgcggatgac acatcgattt acatgttcta tcaaaaagtc ggcgaaactt
841 ctattgacag ctggaaaaac gctggccgcg tctttaaaga cagcgacaaa ttcgatgcaa
901 atgattctat cctaaaagac caaacacaag aatggtcagg ttcagccaca tttacatctg
961 acggaaaaat ccgtttattc tacactgatt tctccggtaa acattacggc aaacaaacac
1021 tgacaactgc acaagttaac gtatcagcat cagacagctc tttgaacatc aacggtgtag
1081 aggattataa atcaatcttt gacggtgacg gaaaaacgta tcaaaatgta cagcagttca
1141 tcgatgaagg caactacagc tcaggcgaca accatacgct gagagatcct cactacgtag
1201 aagataaagg ccacaaatac ttagtatttg aagcaaacac tggaactgaa gatggctacc
1261 aaggcgaaga atctttattt aacaaagcat actatggcaa aagcacatca ttcttccgtc
1321 aagaaagtca aaaacttctg caaagcgata aaaaacgcac ggctgagtta gcaaacggcg
1381 ctctcggtat gattgagcta aacgatgatt acacactgaa aaaagtgatg aaaccgctga
1441 ttgcatctaa cacagtaaca gatgaaattg aacgcgcgaa cgtctttaaa atgaacggca
1501 aatggtacct gttcactgac tcccgcggat caaaaatgac gattgacggc attacgtcta
1561 acgatattta catgcttggt tatgtttcta attctttaac tggcccatac aagccgctga
1621 acaaaactgg ccttgtgtta aaaatggatc ttgatcctaa cgatgtaacc tttacttact
1681 cacacttcgc tgtacctcaa gcgaaaggaa acaatgtcgt gattacaagc tatatgacaa
1741 acagaggatt ctacgcagac aaacaatcaa cgtttgcgcc aagcttcctg ctgaacatca
1801 aaggcaagaa aacatctgtt gtcaaagaca gcatccttga acaaggacaa ttaacagtta
1861 acaaataaaa acgcaaaaga aaatgccgat atcctattgg cattttcttt tatttcttat
1921 caacataaag gtgaatccca tatgaactat ataaaagcag gcaaatggct aaccgtattc
1981 ctaacctttt ggtaatgact ccaacttatt gatagtgttt tatgttcaga taatgcccga
2041 tgactttgtc atgcagcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
2101 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
2161 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
2221 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
2281 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
2341 gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
2401 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
2461 cgatgtaacc cactcgtgca ctttacactt tatgcttccg gctcgtatgt taggaggtct
2521 ttatcatgtc aatttatcaa gaatttgtta ataaatatag tttaagtaaa actctaagat
2581 ttgagttaat cccacagggt aaaacacttg aaaacataaa agcaagaggt ttgattttag
2641 atgatgagaa aagagctaaa gactacaaaa aggctaaaca aataattgat aaatatcatc
2701 agttttttat agaggagata ttaagttcgg tttgtattag cgaagattta ttacaaaact
2761 attctgatgt ttattttaaa cttaaaaaga gtgatgatga taatctacaa aaagatttta
2821 aaagtgcaaa agatacgata aagaaacaaa tatctgaata tataaaggac tcagagaaat
2881 ttaagaattt gtttaatcaa aaccttatcg atgctaaaaa agggcaagag tcagatttaa
2941 ttctatggct aaagcaatct aaggataatg gtatagaact atttaaagcc aatagtgata
3001 tcacagatat agatgaggcg ttagaaataa tcaaatcttt taaaggttgg acaacttatt
3061 ttaagggttt tcatgaaaat agaaaaaatg tttatagtag caatgatatt cctacatcta
3121 ttatttatag gatagtagat gataatttgc ctaaatttct agaaaataaa gctaagtatg
3181 agagtttaaa agacaaagct ccagaagcta taaactatga acaaattaaa aaagatttgg
3241 cagaagagct aacctttgat attgactaca aaacatctga agttaatcaa agagtttttt
3301 cacttgatga agtttttgag atagcaaact ttaataatta tctaaatcaa agtggtatta
3361 ctaaatttaa tactattatt ggtggtaaat ttgtaaatgg tgaaaataca aagagaaaag
3421 gtataaatga atatataaat ctatactcac agcaaataaa tgataaaaca ctcaaaaaat
3481 ataaaatgag tgttttattt aagcaaattt taagtgatac agaatctaaa tcttttgtaa
3541 ttgataagtt agaagatgat agtgatgtag ttacaacgat gcaaagtttt tatgagcaaa
3601 tagcagcttt taaaacagta gaagaaaaat ctattaaaga aacactatct ttattatttg
3661 atgatttaaa agctcaaaaa cttgatttga gtaaaattta ttttaaaaat gataaatctc
3721 ttactgatct atcacaacaa gtttttgatg attatagtgt tattggtaca gcggtactag
3781 aatatataac tcaacaaata gcacctaaaa atcttgataa ccctagtaag aaagagcaag
3841 aattaatagc caaaaaaact gaaaaagcaa aatacttatc tctagaaact ataaagcttg
3901 ccttagaaga atttaataag catagagata tagataaaca gtgtaggttt gaagaaatac
3961 ttgcaaactt tgcggctatt ccgatgatat ttgatgaaat agctcaaaac aaagacaatt
4021 tggcacagat atctatcaaa tatcaaaatc aaggtaaaaa agacctactt caagctagtg
4081 cggaagatga tgttaaagct atcaaggatc ttttagatca aactaataat ctcttacata
4141 aactaaaaat atttcatatt agtcagtcag aagataaggc aaatatttta gacaaggatg
4201 agcattttta tctagtattt gaggagtgct actttgagct agcgaatata gtgcctcttt
4261 ataacaaaat tagaaactat ataactcaaa agccatatag tgatgagaaa tttaagctca
4321 attttgagaa ctcgactttg gctaatggtt gggataaaaa taaagagcct gacaatacgg
4381 caattttatt tatcaaagat gataaatatt atctgggtgt gatgaataag aaaaataaca
4441 aaatatttga tgataaagct atcaaagaaa ataaaggcga gggttataaa aaaattgttt
4501 ataaactttt acctggcgca aataaaatgt tacctaaggt tttcttttct gctaaatcta
4561 taaaatttta taatcctagt gaagatatac ttagaataag aaatcattcc acacatacaa
4621 aaaatggtag tcctcaaaaa ggatatgaaa aatttgagtt taatattgaa gattgccgaa
4681 aatttataga tttttataaa cagtctataa gtaagcatcc ggagtggaaa gattttggat
4741 ttagattttc tgatactcaa agatataatt ctatagatga attttataga gaagttgaaa
4801 atcaaggcta caaactaact tttgaaaata tatcagagag ctatattgat agcgtagtta
4861 atcagggtaa attgtaccta ttccaaatct ataataaaga tttttcagct tatagcaaag
4921 ggcgaccaaa tctacatact ttatattgga aagcgctgtt tgatgagaga aatcttcaag
4981 atgtggttta taagctaaat ggtgaggcag agctttttta tcgtaaacaa tcaataccta
5041 aaaaaatcac tcacccagct aaagaggcaa tagctaataa aaacaaagat aatcctaaaa
5101 aagagagtgt ttttgaatat gatttaatca aagataaacg ctttactgaa gataagtttt
5161 tctttcactg tcctattaca atcaatttta aatctagtgg agctaataag tttaatgatg
5221 aaatcaattt attgctaaaa gaaaaagcaa atgatgttca tatattaagt atagatagag
5281 gtgaaagaca tttagcttac tatactttgg tagatggtaa aggcaatatc atcaaacaag
5341 atactttcaa catcattggt aatgatagaa tgaaaacaaa ctaccatgat aagcttgctg
5401 caatagagaa agatagggat tcagctagga aagactggaa aaagataaat aacatcaaag
5461 agatgaaaga gggctatcta tctcaggtag ttcatgaaat agctaagcta gttatagagt
5521 ataatgctat tgtggttttt gaggatttaa attttggatt taaaagaggg cgtttcaagg
5581 tagagaagca ggtctatcaa aagttagaaa aaatgctaat tgagaaacta aactatctag
5641 ttttcaaaga taatgagttt gataaaactg ggggagtgct tagagcttat cagctaacag
5701 caccttttga gacttttaaa aagatgggta aacaaacagg tattatctac tatgtaccag
5761 ctggttttac ttcaaaaatt tgtcctgtaa ctggttttgt aaatcagtta tatcctaagt
5821 atgaaagtgt cagcaaatct caagagttct ttagtaagtt tgacaagatt tgttataacc
5881 ttgataaggg ctattttgag tttagttttg attataaaaa ctttggtgac aaggctgcca
5941 aaggcaagtg gactatagct agctttggga gtagattgat taactttaga aattcagata
6001 aaaatcataa ttgggatact cgagaagttt atccaactaa agagttggag aaattgctaa
6061 aagattattc tatcgaatat gggcatggcg aatgtatcaa agcagctatt tgcggtgaga
6121 gcgacaaaaa gttttttgct aagctaacta gtgtcctaaa tactatctta caaatgcgta
6181 actcaaaaac aggtactgag ttagattatc taatttcacc agtagcagat gtaaatggca
6241 atttctttga ttcgcgacag gcgccaaaaa atatgcctca agatgctgat gccaatggtg
6301 cttatcatat tgggctaaaa ggtctgatgc tactaggtag gatcaaaaat aatcaagagg
6361 gcaaaaaact caatttggtt atcaaaaatg aagagtattt tgagttcgtg cagaatagga
6421 ataactaaga attcttgaca gctagctcag tcctaggtat aatgctagcg ctgatttagg
6481 caaaaacggg tctaagaact ttaaataatt tctactgttg tagatgagag caggtgtcag
6541 ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg
6601 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
6661 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc
6721 gcgtaatacg actcactata gggcgaattg ggtaccgggc cccccctcga ggtcctccag
6781 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
6841 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
6901 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
6961 gcccgctttc cacccacctg caaaagtcta agaactttaa ataatttcta ctgttgtaga
7021 tgctactatt cctgtgcctt cagataattc agtctaagaa ctttaaataa tttctactgt
7081 tgtagatgtc tagagccttt tgtattagta gccggtctaa gaactttaaa taatttctac
7141 tgttgtagat tagcgattta tgaaggtcat ttttttgtct agctttaatg cggtagttgg
7201 taccgtcgat ggcagaaatt cgatatctag atctcatgga tccgcaatgg caacaacgtt
7261 gcgctcgact ctagaagacc gaaaccttgc gctcgttcgc cagccaggac agaaatgcct
7321 cgacttcgct gctgcccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac
7381 gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct
7441 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
7501 tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc
7561 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc
7621 agggcagtcg ccctaaaaca aagttaaaca tcatgaggga agcggtgatc gccgaagtat
7681 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg
7741 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt
7801 tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc
7861 ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca
7921 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg
7981 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg
8041 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg
8101 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct
8161 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt
8221 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg
8281 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg
8341 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg
8401 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataatgtcta acaattcgtt
8461 caagccgacg ccgcttcgcg gcgcggctta actcaagcgt tagatgcact aagcacataa
8521 ttgctcacag ccaaactatc aggtcaagtc tgcttttatt atttttaagc gtgcataata
8581 agccctacac aaattgggag atatatcatg aaaggctggc tttttcttgt tatcgcaata
8641 gttggcgaag taatcgcaac atccgcatta aaatctagcg agggctttac taagctgatc
8701 cggtggatga ccttttgaat gacctttaat agattatatt actaattaat tggggaccct
8761 agaggtcccc ttttttattt taaaaatttt ttcacaaaac ggtttacaag cataaagctt
8821 gctcaatctg aaagcgacca ggtgctcggc gtggcaagac tcgcagcgaa cccgtagaaa
8881 gccatgctcc agccgcccgc attggagaaa ttcttcaaat tcccgttgca catagcccgg
8941 caattccttt ccctgctctg ccataagcgc agcgaatgcc gggtaatact cgtcaacgat
9001 ctgatagaga agggtttgct cgggtcggtg gctctggtaa cgaccagtat cccgatcccg
9061 gctggccgtc ctggccgcca catgaggcat gttccgcgtc cttgcaatac tgtgtttaca
9121 tacagtctat cgcttagcgg aaagttcttt taccctcagc cgaaatgcct gccgttgcta
9181 gacattgcca gccagtgccc gtcactcccg tactaactgt cacgaacccc tgcaataact
9241 gtcacgcccc cctgcaataa ctgtcacgaa cccctgcaat aactgtcacg cccccaaacc
9301 tgcaaaccca gcaggggcgg gggctggcgg ggtgttggaa aaatccatcc atgattatct
9361 aagaataatc cactaggcgc ggttatcagc gcccttgtgg ggcgctgctg cccttgccca
9421 atatgcccgg ccagaggccg gatagctggt ctattcgctg cgctaggcta cacaccgccc
9481 caccgctgcg cggcaggggg aaaggcgggc aaagcccgct aaaccccaca ccaaaccccg
9541 cagaaatacg ctggagcgct tttagccgct ttagcggcct ttccccctac ccgaagggtg
9601 ggggcgcgtg tgcagccccg cagggcctgt ctcggtcgat cattcagccc ggctcatcct
9661 tctggcgtgg cggcagaccg aacaaggcgc ggtcgtggtc gcgttcaagg tacgcatcca
9721 ttgccgccat gagccgatcc tccggccact cgctgctgtt caccttggcc aaaatcatgg
9781 cccccaccag caccttgcgc cttgtttcgt tcttgcgctc ttgctgctgt tcccttgccc
9841 gcacccgctg aatttcggca ttgattcgcg ctcgttgttc ttcgagcttg gccagccgat
9901 ccgccgcctt gttgctcccc ttaaccatct tgacacccca ttgttaatgt gctgtctcgt
9961 aggctatcat ggaggcacag cggcggcaat cccgacccta ctttgtaggg gagggcgcac
10021 ttaccggttt ctcttcgaga aactggccta acggccaccc ttcgggcggt gcgctctccg
10081 agggccattg catggagccg aaaagcaaaa gcaacagcga ggcagcatgg cgatttatca
10141 ccttacggcg aaaaccggca gcaggtcggg cggccaatcg gccagggcca aggccgacta
10201 catccagcgc gaaggcaagt atgcccgcga catggatgaa gtcttgcacg ccgaatccgg
10261 gcacatgccg gagttcgtcg agcggcccgc cgactactgg gatgctgccg acctgtatga
10321 acgcgccaat gggcggctgt tcaaggaggt cgaatttgcc ctgccggtcg agctgaccct
10381 cgaccagcag aaggcgctgg cgtccgagtt cgcccagcac ctgaccggtg ccgagcgcct
10441 gccgtatacg ctggccatcc atgccggtgg cggcgagaac ccgcactgcc acctgatgat
10501 ctccgagcgg atcaatgacg gcatcgagcg gcccgccgct cagtggttca agcggtacaa
10561 cggcaagacc ccggagaagg gcggggcaca gaagaccgaa gcgctcaagc ccaaggcatg
10621 gcttgagcag acccgcgagg catgggccga ccatgccaac cgggcattag agcgggctgg
10681 ccacgacgcc cgcattgacc acagaacact tgaggcgcag ggcatcgagc gcctgcccgg
10741 tgttcacctg gggccgaacg tggtggagat ggaaggccgg ggcatccgca ccgaccgggc
10801 agacgtggcc ctgaacatcg acaccgccaa cgcccagatc atcgacttac aggaataccg
10861 ggaggcaata gaccatgaac gcaatcgaca gagtgaagaa atccagaggc atcaacgagt
10921 tagcggagca gatcgaaccg ctggcccaga gcatggcgac actggccgac gaagcccggc
10981 aggtcatgag ccagacccag caggccagcg aggcgcaggc ggcggagtgg ctgaaagccc
11041 agcgccagac aggggcggca tgggtggagc tggccaaaga gttgcgggag gtagccgccg
11101 aggtgagcag cgccgcgcag agcgcccgga gcgcgtcgcg ggggtggcac tggaagctat
11161 ggctaaccgt gatgctggct tccatgatgc ctacggtggt gctgctgatc gcatcgttgc
11221 tcttgctcga cctgacgcca ctgacaaccg aggacggctc gatctggctg cgcttggtgg
11281 cccgatgaag aacgacagga ctttgcaggc cataggccga cagctcaagg ccatgggctg
11341 tgagcgcttc gatatcggcg tcagggacgc caccaccggc cagatgatga accgggaatg
11401 gtcagccgcc gaagtgctcc agaacacgcc atggctcaag cggatgaatg cccagggcaa
11461 tgacgtgtat atcaggcccg ccgagcagga gcggcatggt ctggtgctgg tggacgacct
11521 cagcgagttt gacttggatg acatgaaagc cgagggccgg gagcctgccc tggtagtgga
11581 aaccagcccg aagaactatc aggcatgggt caaggtggcc gacgccgcag gcggtgaact
11641 tcgggggcag attgcccgga cgctggccag cgagtacgac gccgacccgg ccagcgccga
11701 cagccgccac tatggccgct tggcgggctt caccaaccgc aaggacaagc acaccacccg
11761 cgccggttat cagccgtggg tgctgctgcg tgaatccaag ggcaagaccg ccaccgctgg
11821 cccggcgctg gtgcagcagg ctggccagca gatcgagcag gcccagcggc agcaggagaa
11881 ggcccgcagg ctggccagcc tcgaactgcc cgagcggcag cttagccgcc accggcgcac
11941 ggcgctggac gagtaccgca gcgagatggc cgggctggtc aagcgcttcg gtgatgacct
12001 cagcaagtgc gactttatcg ccgcgcagaa gctggccagc cggggccgca gtgccgagga
12061 aatcggcaag gccatggccg aggccagccc agcgctggca gagcgcaagc ccggccacga
12121 agcggattac atcgagcgca ccgtcagcaa ggtcatgggt ctgcccagcg tccagcttgc
12181 gcgggccgag ctggcacggg caccggcacc ccgccagcga ggcatggaca ggggcgggcc
12241 agatttcagc atgtagtgct tgcgttggta ctcacgcctg ttatactatg agtactcacg
12301 cacagaaggg ggttttatgg aatacgaaaa aagcgcttca gggtcggtct acctgatcaa
12361 aagtgacaag ggctattggt tgcccggtgg ctttggttat acgtcaaaca aggccgaggc
12421 tggccgcttt tcagtcgctg atatggccag ccttaacctt gacggctgca ccttgtcctt
12481 gttccgcgaa gacaagcctt tcggccccgg caagtttctc ggtgactgat atgaaagacc
12541 aaaaggacaa gcagaccggc gacctgctgg ccagccctga cgctgtacgc caagcgcgat
12601 atgccgagcg catgaaggcc aaagggatgc gtcagcgcaa gttctggctg accgacgacg
12661 aatacgaggc gctgcgcgag tgcctggaag aactcagagc ggcgcagggc gggggtagtg
12721 accccgccag cgcctaacca ccaactgcct gcaaaggagg caatcaatgg ctacccataa
12781 gcctatcaat attctggagg cgttcgcagc agcgccgcca ccgctggact acgttttgcc
12841 caacatggtg gccggtacgg tcggggcgct ggtgtcgccc ggtggtgccg gtaaatccat
12901 gctggccctg caactggccg cacagattgc aggcgggccg gatctgctgg aggtgggcga
12961 actgcccacc ggcccggtga tctacctgcc cgccgaagac ccgcccaccg ccattcatca
13021 ccgcctgcac gcccttgggg cgcacctcag cgccgaggaa cggcaagccg tggctgacgg
13081 cctgctgatc cagccgctga tcggcagcct gcccaacatc atggccccgg agtggttcga
13141 cggcctcaag cgcgccgccg agggccgccg cctgatggtg ctggacacgc tgcgccggtt
13201 ccacatcgag gaagaaaacg ccagcggccc catggcccag gtcatcggtc gcatggaggc
13261 catcgccgcc gataccgggt gctctatcgt gttcctgcac catgccagca agggcgcggc
13321 catgatgggc gcaggcgacc agcagcaggc cagccggggc agctcggtac tggtcgataa
13381 catccgctgg cagtcctacc tgtcgagcat gaccagcgcc gaggccgagg aatggggtgt
13441 ggacgacgac cagcgccggt tcttcgtccg cttcggtgtg agcaaggcca actatggcgc
13501 accgttcgct gatcggtggt tcaggcggca tgacggcggg gtgctcaagc ccgccgtgct
13561 ggagaggcag cgcaagagca agggggtgcc ccgtggtgaa gcctaagaac aagcacagcc
13621 tcagccacgt ccggcacgac ccggcgcact gtctggcccc cggcctgttc cgtgccctca
13681 agcggggcga gcgcaagcgc agcaagctgg acgtgacgta tgactacggc gacggcaagc
13741 ggatcgagtt cagcggcccg gagccgctgg gcgctgatga tctgcgcatc ctgcaagggc
13801 tggtggccat ggctgggcct aatggcctag tgcttggccc ggaacccaag accgaaggcg
13861 gacggcagct ccggctgttc ctggaaccca agtgggaggc cgtcaccgct gatgccatgg
13921 tggtcaaagg tagctatcgg gcgctggcaa aggaaatcgg ggcagaggtc gatagtggtg
13981 gggcgctcaa gcacatacag gactgcatcg agcgcctttg gaaggtatcc atcatcgccc
14041 agaatggccg caagcggcag gggtttcggc tgctgtcgga gtacgccagc gacgaggcgg
14101 acgggcgcct gtacgtggcc ctgaacccct tgatcgcgca ggccgtcatg ggtggcggcc
14161 agcatgtgcg catcagcatg gacgaggtgc gggcgctgga cagcgaaacc gcccgcctgc
14221 tgcaccagcg gctgtgtggc tggatcgacc ccggcaaaac cggcaaggct tccatagata
14281 ccttgtgcgg ctatgtctgg ccgtcagagg ccagtggttc gaccatgcgc aagcgccgcc
14341 agcgggtgcg cgaggcgttg ccggagctgg tcgcgctggg ctggacggta accgagttcg
14401 cggcgggcaa gtacgacatc acccggccca aggcggcagg ctgacccccc ccactctatt
14461 gtaaacaaga catttttatc ttttatattc aatggcttat tttcctgcta attggtaata
14521 ccatgaaaaa taccatgctc agaaaaggct taacaatatt ttgaaaaatt gcctactgag
14581 cgctgccgca cagctccata ggccgctttc ctggctttgc ttccagatgt atgctcttct
14641 gctctagcta gaggatcgat cctttttaac ccat
//

