| 质粒名称: | pGPTVII-Bar-U-MatryoshCaMP6s |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 100024 |
| 存储实验室: | Wolf Frommer |
|
载体骨架基本信息 |
|
| 载体名称: | pGPTVII-Bar-U |
| 载体出品公司: | Clontech |
| 空载体大小: | 11751bp |
| 完整质粒大小: | 13710bp |
| 载体类型: | 植物表达 |
| 筛选标记: | Basta |
| 载体抗性: | Kanamycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | MatryoshCaMP6s、MCaMP6s、GCaMP6s-LSSmOrange |
| 插入大小: | 1959bp |
| 启动子: | AtUBQ10 |
| 克隆方法: | Ligation Independent Cloning |
| 5’克隆位点: | AgeI(未破坏) |
| 3’克隆位点: | NotI (未破坏) |
| 5’测序引物 | CGATTTGTGATTTCTATCTAGATCTGG |
| 3’测序引物 | CACAAACTTAAGCACACAAGC |
|
文献引用方法 |
|
| 材料和方法部分: | pGPTVII-Bar-U-MatryoshCaMP6s was a gift from Wolf Frommer (Addgene plasmid # 100024 ; http://n2t.net/addgene:100024 ; RRID:Addgene_100024) |
| 参考文献部分: | Ratiometric Matryoshka biosensors from a nested cassette of green- and orange-emitting fluorescent proteins. Ast C, Foret J, Oltrogge LM, De Michele R, Kleist TJ, Ho CH, Frommer WB. Nat Commun. 2017 Sep 5;8(1):431. doi: 10.1038/s41467-017-00400-2. 10.1038/s41467-017-00400-2 [pii] PubMed 28874729 |
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 pGPTVII-Bar-U-MatryoshCaMP6s 13710 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Plant expression of fluorescent reporter for calcium signaling,
based off of GCaMP6s. Contains a stable reference Large Stokes Shift
(LSS) mOrange nested within the reporting cpEGFP..
ACCESSION .
VERSION .
KEYWORDS pGPTVII-Bar-U-MatryoshCaMP6s
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13710)
AUTHORS Ast C, Foret J, Oltrogge LM, De Michele R, Kleist TJ, Ho CH, Frommer
WB
TITLE Ratiometric Matryoshka biosensors from a nested cassette of green-
and orange-emitting fluorescent proteins.
JOURNAL Nat Commun. 2017 Sep 5;8(1):431. doi: 10.1038/s41467-017-00400-2.
PUBMED 28874729
REFERENCE 2 (bases 1 to 13710)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..13710
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 1..57
/codon_start=1
/product="calmodulin-binding peptide from avian smooth
muscle myosin light chain kinase"
/label=calmodulin-binding peptide
/translation="RRKWNKTGHAVRAIGRLSS"
primer_bind 270..291
/label=EGFP-C
/note="EGFP, forward primer"
CDS 343..1050
/codon_start=1
/product="monomeric orange fluorescent protein with a large
Stokes shift (Shcherbakova et al., 2012)"
/label=LSSmOrange
/note="mammalian codon-optimized"
/translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
FQTVKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGMEASSERMYPEDGALK
GEDKLRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind 756..775
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind complement(1105..1126)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1366..1385)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
misc_feature 2374..2398
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 2863..3513
/codon_start=1
/gene="tetR"
/product="tetracycline resistance regulatory protein"
/label=TetR
/translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
primer_bind complement(3701..3718)
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(3952..3971)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
oriT 4194..4303
/note="incP origin of transfer"
CDS 4336..4707
/codon_start=1
/gene="traJ"
/product="oriT-recognizing protein"
/label=traJ
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
rep_origin 4834..5545
/label=oriV
/note="incP origin of replication"
mobile_element 6168..6935
/mobile_element_type="insertion sequence:IS1"
/note="prokaryotic transposable element"
CDS 7127..7921
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
CDS 8220..9368
/codon_start=1
/product="trans-acting replication protein that binds to
and activates oriV"
/label=trfA
/translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
misc_feature 10964..10988
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
primer_bind complement(11235..11256)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 11447..11466
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 11591..11613
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 11605..11622
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 11606..11622
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
terminator 11635..11835
/label=gene 7 terminator
/note="transcription terminator for octopine-type Ti
plasmid gene 7"
CDS complement(11870..12421)
/codon_start=1
/gene="Streptomyces hygroscopicus bar"
/product="phosphinothricin acetyltransferase"
/label=BlpR
/note="confers resistance to bialophos or phosphinothricin"
/translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
WQLDFSLPVPPRPVLPVTEI"
promoter complement(12472..12651)
/label=NOS promoter
/note="nopaline synthase promoter"
ORIGIN
1 cgtcgtaagt ggaataagac aggtcacgca gtcagagcta taggtcggct gagctcactc
61 gagaacgtct atatcaaggc cgacaagcag aagaacggca tcaaggcgaa cttccacatc
121 cgccacaaca tcgaggacgg cggcgtgcag ctcgcctacc actaccagca gaacaccccc
181 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcgtgca gtccaaactt
241 tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
301 gggatcactc tcggcatgga cgagctgtac aagggcggta ccatggtgag caagggcgag
361 gagaataaca tggccatcat caaggagttc atgcgcttca aggtgcgcat ggagggctcc
421 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcttt
481 cagaccgtta agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcttg
541 tcccctcagt tcacctacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
601 tacctcaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
661 ggcggcgtgg tgaccgtgac tcaggactcc tccctgcagg acggcgagtt catctacaag
721 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
781 ggcatggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgaggac
841 aagctcaggc tgaagctgaa ggacggcggc cactacacct ccgaggtcaa gaccacctac
901 aaggccaaga agcccgtgca gttgcccggc gcctacatcg tcgacatcaa gttggacatc
961 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
1021 tccaccggcg gcatggacga gctgtacaag ggagggagca tggtgagcaa gggcgaggag
1081 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
1141 ttcagcgtgt ccggcgaggg tgagggcgat gccacctacg gcaagctgac cctgaagttc
1201 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
1261 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
1321 gccatgcccg aaggctacat ccaggagcgc accatcttct tcaaggacga cggcaactac
1381 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
1441 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caacctgccg
1501 gaccaactga ctgaagagca gatcgcagaa tttaaagagg ctttctccct atttgacaag
1561 gacggggatg ggacaataac aaccaaggag ctggggacgg tgatgcggtc tctggggcag
1621 aaccccacag aagcagagct gcaggacatg atcaatgaag tagatgccga cggtgacggc
1681 acaatcgact tccctgagtt cctgacaatg atggcaagaa aaatgaaata cagggacacg
1741 gaagaagaaa ttagagaagc gttcggtgtg tttgataagg atggcaatgg ctacatcagt
1801 gcagcagagc ttcgccacgt gatgacaaac cttggagaga agttaacaga tgaagaggtt
1861 gatgaaatga tcagggaagc agacatcgat ggggatggtc aggtaaacta cgaagagttt
1921 gtacaaatga tgacagcgaa gtgacccggg taggagctca tatgaagatg aagatgaaat
1981 atttggtgtg tcaaataaat agcttgtgtg cttaagtttg tgtttttttc ttggcttgtt
2041 gtgttatgaa tttgtggctt tttctaatat taaatgaatg taagatctca ttataatgaa
2101 taaacaaatg tttctataat ccattgtgaa tgttttgttg gatctcttct gcagcatata
2161 actactgtat gtgctatggt atggactatg gaatatgatt aaagataagg aattcagatc
2221 ggctgagtgg ctccttcaac gttgcggttc tgtcagttcc aaacgtaaaa cggcttgtcc
2281 cgcgtcatcg gcgggggtca taacgtgact cccttaattc tccgctcatg atcagattgt
2341 cgtttcccgc cttcagttta aactatcagt gtttgacagg atatattggc gggtaaacct
2401 aagagaaaag agcgtttatt agaataatcg gatatttaaa agggcgtgaa aaggtttatc
2461 cgttcgtcca tttgtatgtg catgccaacc acagggttcc ccagatctgg cgccggccag
2521 cgagacgagc aagattggcc gccgcccgaa acgatccgac agcgcgccca gcacaggtgc
2581 gcaggcaaat tgcaccaacg catacagcgc cagcagaatg ccatagtggg cggtgacgtc
2641 gttcgagtga accagatcgc gcaggaggcc cggcagcacc ggcataatca ggccgatgcc
2701 gacagcgtcg agcgcgacag tgctcagaat tacgatcagg ggtatgttgg gtttcacgtc
2761 tggcctccgg accagcctcc gctggtccga ttgaacgcgc ggattcttta tcactgataa
2821 gttggtggac atattatgtt tatcagtgat aaagtgtcaa gcatgacaaa gttgcagccg
2881 aatacagtga tccgtgccgc cctggacctg ttgaacgagg tcggcgtaga cggtctgacg
2941 acacgcaaac tggcggaacg gttgggggtt cagcagccgg cgctttactg gcacttcagg
3001 aacaagcggg cgctgctcga cgcactggcc gaagccatgc tggcggagaa tcatacgcat
3061 tcggtgccga gagccgacga cgactggcgc tcatttctga tcgggaatgc ccgcagcttc
3121 aggcaggcgc tgctcgccta ccgcgatggc gcgcgcatcc atgccggcac gcgaccgggc
3181 gcaccgcaga tggaaacggc cgacgcgcag cttcgcttcc tctgcgaggc gggtttttcg
3241 gccggggacg ccgtcaatgc gctgatgaca atcagctact tcactgttgg ggccgtgctt
3301 gaggagcagg ccggcgacag cgatgccggc gagcgcggcg gcaccgttga acaggctccg
3361 ctctcgccgc tgttgcgggc cgcgatagac gccttcgacg aagccggtcc ggacgcagcg
3421 ttcgagcagg gactcgcggt gattgtcgat ggattggcga aaaggaggct cgttgtcagg
3481 aacgttgaag gaccgagaaa gggtgacgat tgatcaggac cgctgccgga gcgcaaccca
3541 ctcactacag cagagccatg tagacaacat cccctccccc tttccaccgc gtcagacgcc
3601 cgtagcagcc cgctacgggc tttttcatgc cctgccctag cgtccaagcc tcacggccgc
3661 gctcggcctc tctggcggcc ttctggcgct cttccgcttc ctcgctcact gactcgctgc
3721 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
3781 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
3841 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
3901 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
3961 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
4021 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct tttccgctgc ataaccctgc
4081 ttcggggtca ttatagcgat tttttcggta tatccatcct ttttcgcacg atatacagga
4141 ttttgccaaa gggttcgtgt agactttcct tggtgtatcc aacggcgtca gccgggcagg
4201 ataggtgaag taggcccacc cgcgagcggg tgttccttct tcactgtccc ttattcgcac
4261 ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc cggctaccgc cggcgtaaca
4321 gatgagggca agcggatggc tgatgaaacc aagccaacca ggaagggcag cccacctatc
4381 aaggtgtact gccttccaga cgaacgaaga gcgattgagg aaaaggcggc ggcggccggc
4441 atgagcctgt cggcctacct gctggccgtc ggccagggct acaaaatcac gggcgtcgtg
4501 gactatgagc acgtccgcga gctggcccgc atcaatggcg acctgggccg cctgggcggc
4561 ctgctgaaac tctggctcac cgacgacccg cgcacggcgc ggttcggtga tgccacgatc
4621 ctcgccctgc tggcgaagat cgaagagaag caggacgagc ttggcaaggt catgatgggc
4681 gtggtccgcc cgagggcaga gccatgactt ttttagccgc taaaacggcc ggggggtgcg
4741 cgtgattgcc aagcacgtcc ccatgcgctc catcaagaag agcgacttcg cggagctggt
4801 gaagtacatc accgacgagc aaggcaagac cgagcgcctt tccgacgctc accgggctgg
4861 ttgccctcgc cgctgggctg gcggccgtct atggccctgc aaacgcgcca gaaacgccgt
4921 cgaagccgtg tgcgagacac cgcggccgcc ggcgttgtgg atacctcgcg gaaaacttgg
4981 ccctcactga cagatgaggg gcggacgttg acacttgagg ggccgactca cccggcgcgg
5041 cgttgacaga tgaggggcag gctcgatttc ggccggcgac gtggagctgg ccagcctcgc
5101 aaatcggcga aaacgcctga ttttacgcga gtttcccaca gatgatgtgg acaagcctgg
5161 ggataagtgc cctgcggtat tgacacttga ggggcgcgac tactgacaga tgaggggcgc
5221 gatccttgac acttgagggg cagagtgctg acagatgagg ggcgcaccta ttgacatttg
5281 aggggctgtc cacaggcaga aaatccagca tttgcaaggg tttccgcccg tttttcggcc
5341 accgctaacc tgtcttttaa cctgctttta aaccaatatt tataaacctt gtttttaacc
5401 agggctgcgc cctgtgcgcg tgaccgcgca cgccgaaggg gggtgccccc ccttctcgaa
5461 ccctcccggc ccgctaacgc gggcctccca tccccccagg ggctgcgccc ctcggccgcg
5521 aacggcctca ccccaaaaat ggcagcgctg gcagtccttg ccattgccgg gatcggggca
5581 gtaacgggat gggcgatcag cccgagcgcg acgcccggaa gcattgacgt gccgcaggtg
5641 ctggcatcga cattcagcga ccaggtgccg ggcagtgagg gcggcggcct gggtggcggc
5701 ctgcccttca cttcggccgt cggggcattc acggacttca tggcggggcc ggcaattttt
5761 accttgggca ttcttggcat agtggtcgcg ggtgccgtgc tcgtgttcgg gggtgcgata
5821 aacccagcga accatttgag gtgataggta agattatacc gaggtatgaa aacgagaatt
5881 ggacctttac agaattactc tatgaagcgc catatttaaa aagctaccaa gacgaagagg
5941 atgaagagga tgaggaggca gattgccttg aatatattga caatactgat aagataatat
6001 atcttttata tagaagatat cgccgtatgt aaggatttca gggggcaagg cataggcagc
6061 gcgcttatca atatatctat agaatgggca aagcataaaa acttgcatgg actaatgctt
6121 gaaacccagg acaataacct tatagcttgt aaattctatc ataattgggt aatgactcca
6181 acttattgat agtgttttat gttcagataa tgcccgatga ctttgtcatg cagctccacc
6241 gattttgaga acgacagcga cttccgtccc agccgtgcca ggtgctgcct cagattcagg
6301 ttatgccgct caattcgctg cgtatatcgc ttgctgatta cgtgcagctt tcccttcagg
6361 cgggattcat acagcggcca gccatccgtc atccatatca ccacgtcaaa gggtgacagc
6421 aggctcataa gacgccccag cgtcgccata gtgcgttcac cgaatacgtg cgcaacaacc
6481 gtcttccgga gactgtcata cgcgtaaaac agccagcgct ggcgcgattt agccccgaca
6541 tagccccact gttcgtccat ttccgcgcag acgatgacgt cactgcccgg ctgtatgcgc
6601 gaggttaccg actgcggcct gagtttttta agtgacgtaa aatcgtgttg aggccaacgc
6661 ccataatgcg ggctgttgcc cggcatccaa cgccattcat ggccatatca atgattttct
6721 ggtgcgtacc gggttgagaa gcggtgtaag tgaactgcag ttgccatgtt ttacggcagt
6781 gagagcagag atagcgctga tgtccggcgg tgcttttgcc gttacgcacc accccgtcag
6841 tagctgaaca ggagggacag ctgatagaaa cagaagccac tggagcacct caaaaacacc
6901 atcatacact aaatcagtaa gttggcagca tcacccataa ttgtggtttc aaaatcggct
6961 ccgtcgatac tatgttatac gccaactttg aaaacaactt tgaaaaagct gttttctggt
7021 atttaaggtt ttagaatgca aggaacagtg aattggagtt cgtcttgtta taattagctt
7081 cttggggtat ctttaaatac tgtagaaaag aggaaggaaa taataaatgg ctaaaatgag
7141 aatatcaccg gaattgaaaa aactgatcga aaaataccgc tgcgtaaaag atacggaagg
7201 aatgtctcct gctaaggtat ataagctggt gggagaaaat gaaaacctat atttaaaaat
7261 gacggacagc cggtataaag ggaccaccta tgatgtggaa cgggaaaagg acatgatgct
7321 atggctggaa ggaaagctgc ctgttccaaa ggtcctgcac tttgaacggc atgatggctg
7381 gagcaatctg ctcatgagtg aggccgatgg cgtcctttgc tcggaagagt atgaagatga
7441 acaaagccct gaaaagatta tcgagctgta tgcggagtgc atcaggctct ttcactccat
7501 cgacatatcg gattgtccct atacgaatag cttagacagc cgcttagccg aattggatta
7561 cttactgaat aacgatctgg ccgatgtgga ttgcgaaaac tgggaagaag acactccatt
7621 taaagatccg cgcgagctgt atgatttttt aaagacggaa aagcccgaag aggaacttgt
7681 cttttcccac ggcgacctgg gagacagcaa catctttgtg aaagatggca aagtaagtgg
7741 ctttattgat cttgggagaa gcggcagggc ggacaagtgg tatgacattg ccttctgcgt
7801 ccggtcgatc agggaggata tcggggaaga acagtatgtc gagctatttt ttgacttact
7861 ggggatcaag cctgattggg agaaaataaa atattatatt ttactggatg aattgtttta
7921 gtacctagat gtggcgcaac gatgccggcg acaagcagga gcgcaccgac ttcttccgca
7981 tcaagtgttt tggctctcag gccgaggccc acggcaagta tttgggcaag gggtcgctgg
8041 tattcgtgca gggcaagatt cggaatacca agtacgagaa ggacggccag acggtctacg
8101 ggaccgactt cattgccgat aaggtggatt atctggacac caaggcacca ggcgggtcaa
8161 atcaggaata agggcacatt gccccggcgt gagtcggggc aatcccgcaa ggagggtgaa
8221 tgaatcggac gtttgaccgg aaggcataca ggcaagaact gatcgacgcg gggttttccg
8281 ccgaggatgc cgaaaccatc gcaagccgca ccgtcatgcg tgcgccccgc gaaaccttcc
8341 agtccgtcgg ctcgatggtc cagcaagcta cggccaagat cgagcgcgac agcgtgcaac
8401 tggctccccc tgccctgccc gcgccatcgg ccgccgtgga gcgttcgcgt cgtctcgaac
8461 aggaggcggc aggtttggcg aagtcgatga ccatcgacac gcgaggaact atgacgacca
8521 agaagcgaaa aaccgccggc gaggacctgg caaaacaggt cagcgaggcc aagcaggccg
8581 cgttgctgaa acacacgaag cagcagatca aggaaatgca gctttccttg ttcgatattg
8641 cgccgtggcc ggacacgatg cgagcgatgc caaacgacac ggcccgctct gccctgttca
8701 ccacgcgcaa caagaaaatc ccgcgcgagg cgctgcaaaa caaggtcatt ttccacgtca
8761 acaaggacgt gaagatcacc tacaccggcg tcgagctgcg ggccgacgat gacgaactgg
8821 tgtggcagca ggtgttggag tacgcgaagc gcacccctat cggcgagccg atcaccttca
8881 cgttctacga gctttgccag gacctgggct ggtcgatcaa tggccggtat tacacgaagg
8941 ccgaggaatg cctgtcgcgc ctacaggcga cggcgatggg cttcacgtcc gaccgcgttg
9001 ggcacctgga atcggtgtcg ctgctgcacc gcttccgcgt cctggaccgt ggcaagaaaa
9061 cgtcccgttg ccaggtcctg atcgacgagg aaatcgtcgt gctgtttgct ggcgaccact
9121 acacgaaatt catatgggag aagtaccgca agctgtcgcc gacggcccga cggatgttcg
9181 actatttcag ctcgcaccgg gagccgtacc cgctcaagct ggaaaccttc cgcctcatgt
9241 gcggatcgga ttccacccgc gtgaagaagt ggcgcgagca ggtcggcgaa gcctgcgaag
9301 agttgcgagg cagcggcctg gtggaacacg cctgggtcaa tgatgacctg gtgcattgca
9361 aacgctaggg ccttgtgggg tcagttccgg ctgggggttc agcagccagc gctttactgg
9421 catttcagga acaagcgggc actgctcgac gcacttgctt cgctcagtat cgctcgggac
9481 gcacggcgcg ctctacgaac tgccgataaa cagaggatta aaattgacaa ttgtgattaa
9541 ggctcagatt cgacggcttg gagcggccga cgtgcaggat ttccgcgaga tccgattgtc
9601 ggccctgaag aaagctccag agatgttcgg gtccgtttac gagcacgagg agaaaaagcc
9661 catggaggcg ttcgctgaac ggttgcgaga tgccgtggca ttcggcgcct acatcgacgg
9721 cgagatcatt gggctgtcgg tcttcaaaca ggaggacggc cccaaggacg ctcacaaggc
9781 gcatctgtcc ggcgttttcg tggagcccga acagcgaggc cgaggggtcg ccggtatgct
9841 gctgcgggcg ttgccggcgg gtttattgct cgtgatgatc gtccgacaga ttccaacggg
9901 aatctggtgg atgcgcatct tcatcctcgg cgcacttaat atttcgctat tctggagctt
9961 gttgtttatt tcggtctacc gcctgccggg cggggtcgcg gcgacggtag gcgctgtgca
10021 gccgctgatg gtcgtgttca tctctgccgc tctgctaggt agcccgatac gattgatggc
10081 ggtcctgggg gctatttgcg gaactgcggg cgtggcgctg ttggtgttga caccaaacgc
10141 agcgctagat cctgtcggcg tcgcagcggg cctggcgggg gcggtttcca tggcgttcgg
10201 aaccgtgctg acccgcaagt ggcaacctcc cgtgcctctg ctcaccttta ccgcctggca
10261 actggcggcc ggaggacttc tgctcgttcc agtagcttta gtgtttgatc cgccaatccc
10321 gatgcctaca ggaaccaatg ttctcggcct ggcgtggctc ggcctgatcg gagcgggttt
10381 aacctacttc ctttggttcc gggggatctc gcgactcgaa cctacagttg tttccttact
10441 gggctttctc agccccagat ctggggtcga tcagccgggg atgcatcagg ccgacagtcg
10501 gaacttcggg tccccgacct gtaccattcg gtgagcaatg gataggggag ttgatatcgt
10561 caacgttcac ttctaaagaa atagcgccac tcagcttcct cagcggcttt atccagcgat
10621 ttcctattat gtcggcatag ttctcaagat cgacagcctg tcacggttaa gcgagaaatg
10681 aataagaagg ctgataattc ggatctctgc gagggagatg atatttgatc acaggcagca
10741 acgctctgtc atcgttacaa tcaacatgct accctccgcg agatcatccg tgtttcaaac
10801 ccggcagctt agttgccgtt cttccgaata gcatcggtaa catgagcaaa gtctgccgcc
10861 ttacaacggc tctcccgctg acgccgtccc ggactgatgg gctgcctgta tcgagtggtg
10921 attttgtgcc gagctgccgg tcggggagct gttggctggc tggtggcagg atatattgtg
10981 gtgtaaacaa attgacgctt agacaactta ataacacatt gcggacgttt ttaatgtact
11041 ggggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
11101 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
11161 ttgatggtgg ttccgaaatc ggcaaaatcc cttataaatc aaaagaatag cccgagatag
11221 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
11281 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcacccaaat
11341 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
11401 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
11461 aaggagcggg cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
11521 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
11581 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattaat tcccatcttg
11641 aaagaaatat agtttaaata tttattgata aaataacaag tcaggtatta tagtccaagc
11701 aaaaacataa atttattgat gcaagtttaa attcagaaat atttcaataa ctgattatat
11761 cagctggtac attgccgtag atgaaagact gagtgcgata ttatgtgtaa tacataaatt
11821 gatgatatag ctagcttagc tcatcggggg atcgatcctc taggggtcat cagatctcgg
11881 tgacgggcag gaccggacgg ggcggtaccg gcaggctgaa gtccagctgc cagaaaccca
11941 cgtcatgcca gttcccgtgc ttgaagccgg ccgcccgcag catgccgcgg ggggcatatc
12001 cgagcgcctc gtgcatgcgc acgctcgggt cgttgggcag cccgatgaca gcgaccacgc
12061 tcttgaagcc ctgtgcctcc agggacttca gcaggtgggt gtagagcgtg gagcccagtc
12121 ccgtccgctg gtggcggggg gagacgtaca cggtcgactc ggccgtccag tcgtaggcgt
12181 tgcgtgcctt ccaggggccc gcgtaggcga tgccggcgac ctcgccgtcc acctcggcga
12241 cgagccaggg atagcgctcc cgcagacgga cgaggtcgtc cgtccactcc tgcggttcct
12301 gcggctcggt acggaagttg accgtgcttg tctcgatgta gtggttgacg atggtgcaga
12361 ccgccggcat gtccgcctcg gtggcacggc ggatgtcggc cgggcgtcgt tctgggctca
12421 tggtagatcc ccctagagtc gagatctgga ttgagagtga atatgagact ctaattggat
12481 accgagggga atttatggaa cgtcagtgga gcatttttga caagaaatat ttgctagctg
12541 atagtgacct taggcgactt ttgaacgcgc aataatggtt tctgacgtat gtgcttagct
12601 cattaaactc cagaaacccg cggctgagtg gctccttcaa cgttgcggtt ctgtcagttc
12661 caaacgtaaa acggcttgtc ccgcgtcatc ggcgggggtc ataacgtgac tcccttaatt
12721 ctccgctcat gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca
12781 gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca attccggttc
12841 gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac tgcaagctac
12901 ctgctttctc tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat
12961 ccggggtcag caccgtttct gcggactggc tttctacgtg ttccgcttcc tttagcagcc
13021 cttgcgccct gagtgcttgc ggcagcgtga agcttcgacg agtcagtaat aaacggcgtc
13081 aaagtggttg cagccggcac acacgagtcg tgtttatcaa ctcaaagcac aaatactttt
13141 cctcaaccta aaaataaggc aattagccaa aaacaacttt gcgtgtaaac aacgctcaat
13201 acacgtgtca ttttattatt agctattgct tcaccgcctt agctttctcg tgacctagtc
13261 gtcctcgtct tttcttcttc ttcttctata aaacaatacc caaagacctc ttcttcttca
13321 caattcagat ttcaatttct caaaatctta aaaactttct ctcaattctc tctaccgtga
13381 tcaaggtaaa tttctgtgtt ccttattctc tcaaaatctt cgattttgtt ttcgttcgat
13441 cccaatttcg tatatgttct ttggtttaga ttctgttaat cttagatcga agacgatttt
13501 ctgggtttga tcgttagata tcatcttaat tctcgattag ggtttcatag atatcatccg
13561 atttgttcaa ataatttgag ttttgtcgaa taattactct tcgatttgtg atttctatct
13621 agatctggtg ttagtttcta gtttgtgcga tcgaatttgt cgattaatct gagtttttct
13681 gattaacaga ctagtatggt cgactcatca
//
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