TRUPATH Triple Gai1

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TRUPATH Triple Gai1载体质粒基本信息

质粒名称: TRUPATH Triple Gai1
出品公司: Addgene
目录编号: 196048
存储实验室: Justin English

载体骨架基本信息
载体名称: pcDNA3.1(+)
载体出品公司: Genscript
空载体大小: 5140bp
完整质粒大小: 9868bp
载体类型: 哺乳细胞表达, Luciferase
筛选标记: 新霉素G418
载体抗性: 氨苄霉素
生长温度: 37 ℃
宿主菌: DH5a
拷贝数: 高拷贝

插入基因信息
插入基因名称1: GAlphai1-RLuc8、GNAI1
物种来源: H. sapiens (human)
插入大小: 2031bp
基因突变: RLuc8 and flanking SGGGGS linkers have been inserted at amino acid position 91 of the alpha subunit
启动子: CMV
克隆方法: 未知
5’克隆位点: AgeI(未破坏)
5’测序引物 gccacgttgtgagttggatag
3’测序引物 TAGAAGGCACAGTCGAGG
插入基因名称2: GGamma9-GFP2、GNG9
物种来源: H. sapiens (human)
插入大小: (bp)939
启动子: CMV
标签融合: GFP2 and GSAG linker (N terminal on insert)
克隆方法: 未知
5’测序引物 gagggcagaggaagtcttc
3’测序引物 GATTTAGGTGACACTATAG
插入基因名称3: GBeta3、GNB3
物种来源: H. sapiens (human)
插入大小: (bp)1020
启动子: CMV
克隆方法: 未知
5’测序引物 CGCAAATGGGCGGTAGGCGTG
3’测序引物 ccggtagggccgggattc

文献引用方法
材料和方法部分: TRUPATH Triple Gai1 was a gift from Justin English (Addgene plasmid # 196048 ; http://n2t.net/addgene:196048 ; RRID:Addgene_196048)
参考文献部分: TRUPATH, an open-source biosensor platform for interrogating the GPCR transducerome. Olsen RHJ, DiBerto JF, English JG, Glaudin AM, Krumm BE, Slocum ST, Che T, Gavin AC, McCorvy JD, Roth BL, Strachan RT. Nat Chem Biol. 2020 May 4. pii: 10.1038/s41589-020-0535-8. doi: 10.1038/s41589-020-0535-8. 10.1038/s41589-020-0535-8 PubMed 32367019

TRUPATH Triple Gai1质粒图谱载体图谱和TRUPATH Triple Gai1载体序列质粒序列多克隆位点信息

TRUPATH

TRUPATH Triple Gai1质粒载体简介

TRUPATH Triple Gai1质粒序列载体序列

LOCUS       TRUPATH Triple Gai1       10159 bp DNA     circular SYN 25-AUG-2024
DEFINITION  Encodes a G alpha subunit (GNAl1) with RLuc8, a G gamma subunit 
            (GNG9) with GFP2 and a G beta subunit (GNB3) as optimal components 
            of a BRET2 biosensor for studying heterotrimeric G proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10159)
  AUTHORS   Olsen RHJ, DiBerto JF, English JG, Glaudin AM, Krumm BE, Slocum ST, 
            Che T, Gavin AC, McCorvy JD, Roth BL, Strachan RT
  TITLE     TRUPATH, an open-source biosensor platform for interrogating the 
            GPCR transducerome.
  JOURNAL   Nat Chem Biol. 2020 May 4. pii: 10.1038/s41589-020-0535-8. doi: 
            10.1038/s41589-020-0535-8.
  PUBMED    32367019
REFERENCE   2  (bases 1 to 10159)
  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: "Nat Chem 
            Biol. 2020 May 4. pii: 10.1038/s41589-020-0535-8. doi: 
            10.1038/s41589-020-0535-8."
FEATURES             Location/Qualifiers
     source          1..10159
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1937..1990
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     complement(3007..3024)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    3036..3609
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3059..3611
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3203..3220)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3430..3449
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(5750..5767)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5756..5980
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6026..6454
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6113..6132)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6323..6344
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6463..6483)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6468..6797
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6648..6783
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6710..6729
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6864..7658
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6918..6937)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7528..7547
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7832..7953
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7869..7888)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7923..7942
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8002..8018)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8002..8018)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8015..8037)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8026..8042
                     /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(8050..8080)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8095..8116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8233..8250)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8404..8992)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8484..8503)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9163..10023)
                     /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     9786..9805
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10024..10128)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttatgg gagagatgga gcagctgagg caggaggcag agcagctgaa
      961 gaagcagatc gccgacgcca gaaaggcctg cgcagatgtg accctggcag agctggtgag
     1021 cggactggag gtggtgggaa gggtgcagat gaggaccagg agaacactga ggggccacct
     1081 ggccaagatc tacgccatgc actgggccac agactctaag ctgctggtgt ccgcctctca
     1141 ggacggcaag ctgatcgtgt gggatagcta taccacaaac aaggtgcacg caatccccct
     1201 gcgcagctcc tgggtcatga cctgtgccta cgccccttcc ggaaatttcg tggcatgcgg
     1261 aggactggat aacatgtgct ctatctataa tctgaagagc cgggagggca acgtgaaggt
     1321 gtctagagag ctgagcgccc acaccggcta cctgtcttgc tgtcggttcc tggacgataa
     1381 caatatcgtg acatctagcg gcgacaccac atgcgcactg tgggacatcg agaccggaca
     1441 gcagaagaca gtgtttgtgg gccacacagg cgactgtatg tccctggccg tgtctccaga
     1501 tttcaatctg tttatctccg gagcatgcga cgcatctgcc aagctgtggg atgtgcggga
     1561 gggcacctgt agacagacct tcacaggcca cgagagcgac atcaacgcca tctgcttctt
     1621 tcccaatggc gaggccatct gcacaggcag cgacgatgcc tcctgtaggc tgttcgacct
     1681 gcgcgccgat caggagctga tctgcttttc tcacgagagc atcatctgtg gcatcacctc
     1741 cgtggccttt agcctgtccg gcaggctgct gttcgcagga tatgacgact tcaactgcaa
     1801 cgtgtgggac tccatgaagt ctgagagggt gggcatcctg agcggccacg ataatcgcgt
     1861 gtcctgtctg ggagtgaccg cagacggaat ggcagtggcc acaggcagct gggattcctt
     1921 tctgaagatc tggaacgagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa
     1981 tcccggccct accggtatgg tgtccaaggg agaggagctg ttcaccggag tggtgcctat
     2041 cctggtggag ctggacggcg atgtgaatgg ccacaagttt agcgtgtccg gagagggaga
     2101 gggcgacgca acctacggca agctgacact gaagttcatc tgcaccacag gcaagctgcc
     2161 cgtgccttgg ccaaccctgg tgaccacact gtcctacggc gtgcagtgtt tttctaggta
     2221 tcctgaccac atgaagcagc acgatttctt taagtctgcc atgccagagg gctacgtgca
     2281 ggagcggaca atcttcttta aggacgatgg caactataag accagagccg aggtgaagtt
     2341 cgagggcgac acactggtga acaggatcga gctgaagggc atcgacttta aggaggatgg
     2401 caatatcctg ggccacaagc tggagtacaa ctataatccc cacaacgtgt atatcatggc
     2461 cgataagcag aagaacggca tcaaggtcaa tttcaagatc cgccacaata tcgaggacgg
     2521 cagcgtgcag ctggccgatc actaccagca gaacacccca atcggcgacg gacccgtgct
     2581 gctgcctgat aatcactatc tgtttacaca gtctgccctg agcaaggacc ccaacgagaa
     2641 gcgggatcac atggtgctgc tggagttcgt gaccgcagca ggcatcacac tgggcatgga
     2701 tgagctgtac aagggcagcg ccggcaccat ggcacaggac ctgtccgaga aggatctgct
     2761 gaagatggag gtggagcagc tgaagaagga ggtgaagaat acaagaatcc ccatcagcaa
     2821 ggccggcaag gagatcaagg agtatgtgga ggcccaggcc ggcaacgacc ctttcctgaa
     2881 gggcatccca gaggataaga atccctttaa ggagaagggc ggctgcatca tcagctgacc
     2941 gcggcgtacg aggcctgcat gctccggacc tgcaggttcg aagtcgacag atctcaattg
     3001 gggcccctat agtgtcacct aaataattcc gcccccccct ctccctcccc cccccctaac
     3061 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     3121 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     3181 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     3241 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     3301 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     3361 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     3421 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     3481 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     3541 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     3601 cgatgataat atgggctgca ccctgtccgc agaggataag gcagcagtgg agcggtctaa
     3661 gatgatcgat cggaatctga gagaggacgg cgagaaggcc gccagagagg tgaagctgct
     3721 gctgctggga gcaggagagt ctggcaagag caccatcgtg aagcagatga agatcatcca
     3781 cgaggccggc tactctgagg aggagtgtaa gcagtacaag gccgtggtgt attctaacac
     3841 aatccagagc atcatcgcca tcatcagggc aatgggaaga agcggaggag gaggctccat
     3901 ggcatctaag gtgtatgacc ctgagcagag gaagcgcatg atcaccggac cacagtggtg
     3961 ggcaaggtgc aagcagatga acgtgctgga ttcctttatc aattactatg actctgagaa
     4021 gcacgccgag aacgccgtga tcttcctgca cggcaatgcc acaagctcct acctgtggag
     4081 gcacgtggtg ccccacatcg agcctgtggc aaggtgtatc atccctgatc tgatcggcat
     4141 gggcaagagc ggcaagtccg gcaatggcag ctataggctg ctggaccact acaagtatct
     4201 gaccgcctgg ttcgagctgc tgaacctgcc aaagaagatc atctttgtgg gacacgactg
     4261 gggagcagca ctggcattcc actacgccta tgagcaccag gatcgcatca aggccatcgt
     4321 gcacatggag agcgtggtgg acgtgatcga gtcctgggac gagtggcctg acatcgagga
     4381 ggacatcgcc ctgatcaagt ctgaggaggg cgagaagatg gtgctggaga acaatttctt
     4441 tgtggagacc gtgctgccta gcaagatcat gaggaagctg gagccagagg agttcgccgc
     4501 ctacctggag ccctttaagg agaagggcga ggtgcggaga cccacactgt cctggccacg
     4561 cgagatccca ctggtgaagg gaggcaagcc tgatgtggtg cagatcgtgc ggaactacaa
     4621 tgcctatctg agagcctctg acgatctgcc aaagctgttc atcgagagcg accccggctt
     4681 cttttccaat gccatcgtgg agggcgccaa gaagttccca aacaccgagt ttgtgaaggt
     4741 gaagggcctg cactttctgc aggaggacgc ccccgatgag atgggcaagt acatcaagag
     4801 cttcgtggag agagtgctga agaatgagca gtccggagga ggaggctctc tgaagatcga
     4861 ctttggcgat tccgccaggg ccgacgatgc aaggcagctg ttcgtgctgg caggagcagc
     4921 agaggagggc tttatgacag cagagctggc aggcgtgatc aagaggctgt ggaaggatag
     4981 cggcgtgcag gcctgcttca accggtctag agagtaccag ctgaatgaca gcgccgccta
     5041 ctatctgaac gacctggata gaatcgccca gcctaactat atcccaaccc agcaggacgt
     5101 gctgaggaca cgcgtgaaga ccacaggcat cgtggagacc cacttcacct tcaaggatct
     5161 gcacttcaag atgtttgacg tgggaggaca gaggtccgag cgcaagaagt ggattcactg
     5221 cttcgagggc gtgaccgcca tcatcttttg tgtggccctg tctgactacg atctggtgct
     5281 ggccgaggat gaggagatga atcggatgca cgagtctatg aagctgttcg atagcatctg
     5341 taacaataag tggtttaccg acacatccat catcctgttc ctgaacaaga aggacctgtt
     5401 tgaggagaag atcaagaaga gccccctgac catctgctac cctgagtatg ccggctccaa
     5461 tacatacgag gaggctgccg cctatatcca gtgtcagttt gaggatctga acaagagaaa
     5521 ggacaccaag gagatctata cccacttcac atgcgccacc gatacaaaga acgtgcagtt
     5581 cgtgtttgat gccgtgacag acgtgatcat caagaacaat ctgaaggact gtggcctgtt
     5641 ctgaggtacc gagctcggat ccactagtcc agtgtggtgg aattctgcag atatccagca
     5701 cagtggcggc cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg
     5761 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     5821 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     5881 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     5941 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     6001 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     6061 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     6121 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     6181 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     6241 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     6301 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     6361 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     6421 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     6481 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     6541 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     6601 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     6661 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     6721 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     6781 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     6841 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     6901 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     6961 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     7021 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     7081 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     7141 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     7201 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     7261 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     7321 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     7381 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     7441 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     7501 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     7561 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     7621 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     7681 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     7741 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     7801 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     7861 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     7921 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     7981 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     8041 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     8101 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     8161 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     8221 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     8281 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     8341 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     8401 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     8461 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     8521 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     8581 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     8641 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     8701 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     8761 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     8821 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     8881 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     8941 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     9001 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     9061 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     9121 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     9181 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     9241 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     9301 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     9361 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     9421 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     9481 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     9541 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     9601 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     9661 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     9721 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     9781 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     9841 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     9901 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     9961 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
    10021 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
    10081 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
    10141 aaaagtgcca cctgacgtc
//

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