pCMV-T7-ABEmax(7.10)-SpRY-P2A-EGFP (RTW5025)

价格:1200元

联系方式:I47-825O-882O

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载体基本信息

质粒名称: pCMV-T7-ABEmax(7.10)-SpRY-P2A-EGFP (RTW5025) 
出品公司: 爱迪基因
目录编号: 140003
存储实验室: Benjamin Kleinstiver

载体骨架基本信息
载体名称: pCMV_ABEmax_P2A_GFP
载体出品公司: David Liu (Addgene Plasmid 112101)
载体抗性: 氨苄青霉素
空载体大小: --
完整质粒大小: --
载体修饰: --
载体类型: 哺乳动物表达,CRISPR;体外转录;T7启动子
筛选标记: --
高拷贝/低拷贝: 高拷贝
生长条件: 37 ℃

插入基因信息
插入基因名称: human codon optimized ABEmax(7.10) SpCas9 variant named SpRY with P2A-EGFP,RTW5025
物种来源: Synthetic
插入大小: --
基因突变: nSpCas9=D10A; SpRY=A61R/L1111R/D1135L/S1136W/G1218K/E1219Q/N1317R/A1322R/R1333P/R1335Q/T1337R
GenBank编号 --
启动子 CMV and T7
标签/融合蛋白: P2A-EGFP(插入件上的C端子)
克隆方法: Gibson克隆
5' 克隆位点: --
3' 克隆位点: --
5' 测序引物 oBK6928-CCAAGTCTCCACCCCATTGACG
3' 测序引物 oBK219-GGGAGTGGCACCTTCCAGGGTC

文献引用方法
材料和方法部分: pCMV-T7-ABEmax(7.10)-SpRY-P2A-EGFP (RTW5025) was a gift from Benjamin Kleinstiver (Addgene plasmid # 140003 ; http://n2t.net/addgene:140003 ; RRID:Addgene_140003)
参考文献部分: Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants. Walton , Russell T., Christie, Kathleen A., Whittaker, Madelynn N., Kleinstiver, Benjamin P.. Science , 26 Mar 2020 10.1126/science.aba8853

载体质粒图谱和多克隆位点信息

pCMV-T7-ABEmax(7.10)-SpRY-P2A-EGFP (RTW5025)质粒图谱

载体简介

载体序列

LOCUS       pCMV-T7-ABEmax(7.10)-SpRY-P2A-EGFP (RTW5025)                9627 bp ds-DNA     circular SYN 20-MAR-2020
DEFINITION  CMV and T7 promoter expression plasmid for human codon optimized 
            ABEmax(7.10) A-to-G base editor with 
            SpRY(D10A/A61R/L1111R/D1135L/S1136W/G1218K/E1219Q/N1317R/A1322R/R133
            3P/R1335Q/T1337R) and P2A-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-T7-ABEmax(7.10)-SpRY-P2A-EGFP (RTW5025)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9627)
  AUTHORS   Kleinstiver
  TITLE     Kleinstiver lab CRISPR plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 9627)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 20, 2020 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9627
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     511..530
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(749..853)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(932..951)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1123..1502
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1503..1706
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1657..1677
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1748..1767
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1748..1766
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1816..1836
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3025..4740
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAERTRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     CDS             7168..7188
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7198..7254
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             7255..7971
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7300..7321)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7561..7580)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7908..7929
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7984..8004
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8013..8030
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(8053..8070)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8059..8283
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8354..8370)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8354..8370)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8367..8389)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8378..8394
                     /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(8402..8432)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8447..8468
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8585..8602)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8756..9344)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8836..8855)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9515..748)
                     /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"
ORIGIN
        1 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
       61 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
      121 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
      181 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
      241 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
      301 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
      361 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
      421 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
      481 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
      541 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
      601 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
      661 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
      721 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
      781 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
      841 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
      901 gatctcccga tcccctaggg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
      961 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     1021 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
     1081 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
     1141 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     1201 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     1261 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     1321 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     1381 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     1441 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     1501 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     1561 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     1621 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     1681 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagagatccg
     1741 cggccgctaa tacgactcac tatagggaga gccgccacca tgaaacggac agccgacgga
     1801 agcgagttcg agtcaccaaa gaagaagcgg aaagtctctg aagtcgagtt tagccacgag
     1861 tattggatga ggcacgcact gaccctggca aagcgagcat gggatgaaag agaagtcccc
     1921 gtgggcgccg tgctggtgca caacaataga gtgatcggag agggatggaa caggccaatc
     1981 ggccgccacg accctaccgc acacgcagag atcatggcac tgaggcaggg aggcctggtc
     2041 atgcagaatt accgcctgat cgatgccacc ctgtatgtga cactggagcc atgcgtgatg
     2101 tgcgcaggag caatgatcca cagcaggatc ggaagagtgg tgttcggagc acgggacgcc
     2161 aagaccggcg cagcaggctc cctgatggat gtgctgcacc accccggcat gaaccaccgg
     2221 gtggagatca cagagggaat cctggcagac gagtgcgccg ccctgctgag cgatttcttt
     2281 agaatgcgga gacaggagat caaggcccag aagaaggcac agagctccac cgactctgga
     2341 ggatctagcg gaggatcctc tggaagcgag acaccaggca caagcgagtc cgccacacca
     2401 gagagctccg gcggctcctc cggaggatcc tctgaggtgg agttttccca cgagtactgg
     2461 atgagacatg ccctgaccct ggccaagagg gcacgcgatg agagggaggt gcctgtggga
     2521 gccgtgctgg tgctgaacaa tagagtgatc ggcgagggct ggaacagagc catcggcctg
     2581 cacgacccaa cagcccatgc cgaaattatg gccctgagac agggcggcct ggtcatgcag
     2641 aactacagac tgattgacgc caccctgtac gtgacattcg agccttgcgt gatgtgcgcc
     2701 ggcgccatga tccactctag gatcggccgc gtggtgtttg gcgtgaggaa cgcaaaaacc
     2761 ggcgccgcag gctccctgat ggacgtgctg cactaccccg gcatgaatca ccgcgtcgaa
     2821 attaccgagg gaatcctggc agatgaatgt gccgccctgc tgtgctattt ctttcggatg
     2881 cctagacagg tgttcaatgc tcagaagaag gcccagagct ccaccgactc cggaggatct
     2941 agcggaggct cctctggctc tgagacacct ggcacaagcg agagcgcaac acctgaaagc
     3001 agcgggggca gcagcggggg gtcagacaag aagtacagca tcggcctggc catcggcacc
     3061 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
     3121 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
     3181 gacagcggcg aaacagccga gagaacccgg ctgaagagaa ccgccagaag aagatacacc
     3241 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
     3301 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
     3361 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
     3421 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
     3481 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
     3541 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
     3601 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
     3661 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
     3721 aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
     3781 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
     3841 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
     3901 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
     3961 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
     4021 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
     4081 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
     4141 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
     4201 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
     4261 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
     4321 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
     4381 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
     4441 accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag
     4501 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
     4561 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
     4621 atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc
     4681 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
     4741 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
     4801 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
     4861 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
     4921 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
     4981 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
     5041 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
     5101 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
     5161 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
     5221 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
     5281 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
     5341 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
     5401 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
     5461 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
     5521 tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac
     5581 aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
     5641 gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc
     5701 cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
     5761 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
     5821 atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg
     5881 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
     5941 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
     6001 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
     6061 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
     6121 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
     6181 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg
     6241 tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat
     6301 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cagacccaag
     6361 aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
     6421 ttcctgtggc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
     6481 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
     6541 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
     6601 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
     6661 ctggcctctg ccaagcagct gcagaaggga aacgaactgg ccctgccctc caaatatgtg
     6721 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
     6781 cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc
     6841 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc
     6901 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
     6961 accctgacca gactgggagc ccctagagcc ttcaagtact ttgacaccac catcgacccc
     7021 aagcagtaca gaagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
     7081 ggcctgtacg agacacggat cgacctgtct cagctgggag gtgactctgg cggctcaaaa
     7141 agaaccgccg acggcagcga attcgagccc aagaagaaga ggaaagtcgg aagcggagct
     7201 actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg acctatggtg
     7261 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     7321 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     7381 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     7441 accaccctga cctatggagt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     7501 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     7561 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     7621 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     7681 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     7741 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     7801 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     7861 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     7921 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtctggtggt
     7981 tctcccaaga agaagaggaa agtctaaccg gtcatcatca ccatcaccat tgagtttaaa
     8041 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     8101 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     8161 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     8221 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     8281 tggcttctga ggcggaaaga accagctggg gctcgatacc gtcgacctct agctagagct
     8341 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     8401 acaacatacg agccggaagc ataaagtgta aagcctaggg tgcctaatga gtgagctaac
     8461 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     8521 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     8581 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     8641 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     8701 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     8761 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     8821 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     8881 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     8941 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     9001 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     9061 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     9121 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     9181 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     9241 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     9301 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     9361 tttctacggg gtctgacact cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     9421 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     9481 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     9541 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     9601 taactacgat acgggagggc ttaccat
//

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