pBAD-Flp

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相关技术服务:质粒构建    基因合成    质粒大提

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pBAD-Flp载体质粒基本信息

质粒名称: pBAD-Flp
出品公司: Addgene
目录编号: 122969
存储实验室: Lydia Freddolino

载体骨架基本信息
载体名称: pCP20
载体出品公司: Clontech
空载体大小: 4038 bp
完整质粒大小: 10691bp
载体类型: 细菌表达
载体抗性: Chloramphenicol, 25 μg/mL
生长温度: 30 ℃
宿主菌: DH5a
拷贝数: 低拷贝

插入基因信息
插入基因名称: Flp recombinase
物种来源: 光合细菌 Rhodopseudomonas palustris
插入大小: 1272bp
启动子: pBAD
克隆方法: Gibson Cloning
5’测序引物
pBAD Forward
(Common Sequencing Primers)

文献引用方法
材料和方法部分: pBAD-Flp was a gift from Lydia Freddolino (Addgene plasmid # 122969 ; http://n2t.net/addgene:122969 ; RRID:Addgene_122969)
参考文献部分: High-Resolution Mapping of the Escherichia coli Chromosome Reveals Positions of High and Low Transcription. Scholz SA, Diao R, Wolfe MB, Fivenson EM, Lin XN, Freddolino PL. Cell Syst. 2019 Mar 27;8(3):212-225.e9. doi: 10.1016/j.cels.2019.02.004. Epub 2019 Mar 20. 10.1016/j.cels.2019.02.004 PubMed 30904377


pBAD-Flp质粒图谱载体图谱和pBAD-Flp载体序列质粒序列多克隆位点信息

pBAD-Flp载体图谱

pBAD-Flp质粒载体简介

pBAD-Flp质粒序列载体序列

LOCUS      pBAD-Flp       10691 bp DNA     circular SYN 25-AUG-2024
DEFINITION  Expresses flp recombinase in bacteria upon arabinose induction. No 
            flp activity is detected without induction..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10691)
  AUTHORS   Scholz SA, Diao R, Wolfe MB, Fivenson EM, Lin XN, Freddolino PL
  TITLE     High-Resolution Mapping of the Escherichia coli Chromosome Reveals 
            Positions of High and Low Transcription.
  JOURNAL   Cell Syst. 2019 Mar 27;8(3):212-225.e9. doi: 
            10.1016/j.cels.2019.02.004. Epub 2019 Mar 20.
  PUBMED    30904377
REFERENCE   2  (bases 1 to 10691)
  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"; doi: 
            "10.1016/j.cels.2019.02.004"; journalName: "Cell Syst"; date: 
            "2019-03-27- 27"; volume: "8"; issue: "3"; pages: "212-225.e9"
FEATURES             Location/Qualifiers
     source          1..10691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1083..1102)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1167..1186)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     CDS             complement(1904..2617)
                     /codon_start=1
                     /gene="cIts"
                     /product="temperature-sensitive variant of the phage lambda
                     repressor"
                     /label=lambda repressor (ts)
                     /note="thermosensitivity is conferred by the A67T mutation"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     CDS             complement(2659..3537)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRH
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3564..3848
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     3771..3790
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3866..5137
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=FLP
                     /note="FLP is a site-specific recombinase from 
                     Saccharomyces cerevisiae. Recombination occurs at FRT 
                     sequences."
                     /translation="MPQFDILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI
                     THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
                     IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
                     SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
                     KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
                     SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
                     YPAWNGIISQEVLDYLSSYINRRI"
     primer_bind     complement(6085..6103)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6171..6275
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6276..7136
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6494..6513)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(8598..9548)
                     /codon_start=1
                     /gene="rep101 encoding an A56V mutation"
                     /product="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /label=Rep101(Ts)
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      9596..9818
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     promoter        10370..10472
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(10473..10691,1..441)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(10539..10558)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
       61 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
      121 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
      181 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
      241 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
      301 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
      361 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
      421 gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggt
      481 tgctacgcct gaataagtga taataagcgg atgaatggca gaaattcgaa agcaaattcg
      541 acccggtcgt cggttcaggg cagggtcgtt aaatagccgc ttatgtctat tgctggttta
      601 ccggtttatt gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc tgaggccagt
      661 ttgctcaggc tctccccgtg gaggtaataa ttgacgatat gatcatttat tctgcctccc
      721 agagcctgat aaaaacggtt agcgccagca accgcacctg tggcgctgag gtctgcctcg
      781 tgaagaaggt gttgctgact cataccaggc ctgaatcgcc ccatcatcca gccagaaagt
      841 gagggagcca cggttgatga gagctttgtt gtaggtggac cagttggtga ttttgaactt
      901 ttgctttgcc acggaacggt ctgcgttgtc gggaagatgc gtgatctgat ccttcaactc
      961 agcaaaagtt cgatttattc aacaaagcca cgttgtgtct caaaatctct gatgttacat
     1021 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     1081 tacaaggggt gttatgagcc atattcaacg ggaaacgtct tgctcgaggc cgcgattaaa
     1141 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     1201 aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     1261 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     1321 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     1381 actcaccact gcgatccccg ggaaaacagc attccaggta ttagaagaat atcctgattc
     1441 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     1501 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     1561 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     1621 acaagtctgg aaagaaatgc agattcttgc tcaattgtta tcagctatgc gccgaccaga
     1681 acaccttgcc gatcagccaa acgtctcttc aggccactga ctagcgataa ctttccccac
     1741 aacggaacaa ctctcattgc atgggatcat tgrrtamcgg tttggttgat gcgagtgatt
     1801 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cagattcttg
     1861 ctcaattgtt atcagctatg cgccgaccag aacaccttgc cgatcagcca aacgtctctt
     1921 caggccactg actagcgata actttcccca caacggaaca actctcattg catgggatca
     1981 ttgggtactg tgggtttagt ggttgtaaaa acacctgacc gctatccctg atcagtttct
     2041 tgaaggtaaa ctcatcaccc ccaagtctgg ctatgcagaa atcacctggc tcaacagcct
     2101 gctcagggtc aacgagaatt aacattccgt caggaaagct tggcttggag cctgttggtg
     2161 cggtcatgga attaccttca acctcaagcc agaatgcaga atcactggct tttttggttg
     2221 tgcttaccca tctctccgca tcacctttgg taaaggttct aagcttaggt gagaacatcc
     2281 ctgcctgaac atgagaaaaa acagggtact catactcact tctaagtgac ggctgcatac
     2341 taaccgcttc atacatctcg tagatttctc tggcgattga agggctaaat tcttcaacgc
     2401 taactttgag aatttttgta agcaatgcgg cgttataagc atttaatgca ttgatgccat
     2461 taaataaagc accaacgcct gactgcccca tccccatctt gtctgcgaca gattcctggg
     2521 ataagccaag ttcatttttc tttttttcat aaattgcttt aaggcgacgt gcgtcctcaa
     2581 gctgctcttg tgttaatggt ttcttttttg tgctcatacg ttaaatctat caccgcaagg
     2641 gataaatatc taacaccgtt atgacaactt gacggctaca tcattcactt tttcttcaca
     2701 accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaagta
     2761 gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct
     2821 caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc
     2881 taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac
     2941 gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg
     3001 tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgt gccgcagtaa
     3061 caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc
     3121 gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa
     3181 gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg
     3241 acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg
     3301 aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg
     3361 atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag
     3421 cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac
     3481 cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact
     3541 tttcatactc ccgccattca gagaagaaac caattgtcca tattgcatca gacattgccg
     3601 tcactgcgtc ttttactggc tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag
     3661 cattctgtaa caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa
     3721 tcacggcaga aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc
     3781 atttttatcc ataagattag cggatcctac ctgacgcttt ttatcgcaac tctctactgt
     3841 ttctccatac ccgttaggag gttgtatgcc acaatttgat atattatgta aaacaccacc
     3901 taaggtgctt gttcgtcagt ttgtggaaag gtttgaaaga ccttcaggtg agaaaatagc
     3961 attatgtgct gctgaactaa cctatttatg ttggatgatt acacataacg gaacagcaat
     4021 caagagagcc acattcatga gctataatac tatcataagc aattcgctga gtttcgatat
     4081 tgtcaataaa tcactccagt ttaaatacaa gacgcaaaaa gcaacaattc tggaagcctc
     4141 attaaagaaa ttgattcctg cttgggaatt tacaattatt ccttactatg gacaaaaaca
     4201 tcaatctgat atcactgata ttgtaagtag tttgcaatta cagttcgaat catcggaaga
     4261 agcagataag ggaaatagcc acagtaaaaa aatgcttaaa gcacttctaa gtgagggtga
     4321 aagcatctgg gagatcactg agaaaatact aaattcgttt gagtatactt cgagatttac
     4381 aaaaacaaaa actttatacc aattcctctt cctagctact ttcatcaatt gtggaagatt
     4441 cagcgatatt aagaacgttg atccgaaatc atttaaatta gtccaaaata agtatctggg
     4501 agtaataatc cagtgtttag tgacagagac aaagacaagc gttagtaggc acatatactt
     4561 ctttagcgca aggggtagga tcgatccact tgtatatttg gatgaatttt tgaggaattc
     4621 tgaaccagtc ctaaaacgag taaataggac cggcaattct tcaagcaata aacaggaata
     4681 ccaattatta aaagataact tagtcagatc gtacaataaa gctttgaaga aaaatgcgcc
     4741 ttattcaatc tttgctataa aaaatggccc aaaatctcac attggaagac atttgatgac
     4801 ctcatttctt tcaatgaagg gcctaacgga gttgactaat gttgtgggaa attggagcga
     4861 taagcgtgct tctgccgtgg ccaggacaac gtatactcat cagataacag caatacctga
     4921 tcactacttc gcactagttt ctcggtacta tgcatatgat ccaatatcaa aggaaatgat
     4981 agcattgaag gatgagacta atccaattga ggagtggcag catatagaac agctaaaggg
     5041 tagtgctgaa ggaagcatac gataccccgc atggaatggg ataatatcac aggaggtact
     5101 agactacctt tcatcctaca taaatagacg catataagta cgcatttaag cataaacacg
     5161 cactatgccg ttcttctcat gtatatatat atacaggcaa cacgcagata taggtgcgac
     5221 gtgaacagtg agctgtatgt gcgcagctcg cgttgcattt tcggaagcgc tcgttttcgg
     5281 aaacgctttg aagttcctat tccgaagttc ctattctgcc attctcaccg gattcagtcg
     5341 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     5401 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     5461 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     5521 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     5581 aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcggct
     5641 ttgttgaata aatcgaactt ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa
     5701 cgcagaccgt tccgtggcaa agcaaaagtt caaaatcacc aactggtcca cctacaacaa
     5761 agctctcatc aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcctg
     5821 gtatgagtca gcaacacctt cttcacgagg cagacctcag cgccggtgat gccggccacg
     5881 atgcgtccgg cgtagaggat cctagagcgc acgaatgagg gccgacagga agcaaagctg
     5941 aaaggaatca aatttggccg caggcgtacc gtggacagga acgtcgtgct gacgcttcat
     6001 cagaagggca ctggtgcaac ggaaattgct catcagctca gtattgcccg ctccacggtt
     6061 tataaaattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     6121 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     6181 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     6241 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     6301 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     6361 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     6421 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     6481 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     6541 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     6601 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     6661 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     6721 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     6781 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
     6841 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     6901 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     6961 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     7021 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     7081 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     7141 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     7201 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     7261 cgttccactg agcgtcagac cccgtttagt cactgatgcc tccgtgtaag ggggatttct
     7321 gttcatgggg taatgatacc gatgaaacgc gagaggatgc tcacaatacg ggttactgat
     7381 gatgaacatg cccggttact tgaacgctgt gagggtaaac aactggcggt atggatgcgg
     7441 cgggtctgcc tgggggggcc ggktgcccgt tccggaaaac tgccgacack ggcaccgccg
     7501 ttactgcgtc agctggccgc catcggaaat aacctgaatc agacagcccg taaggtgaac
     7561 agcgggcagt ggtcttccgg tgaccgggtt caggtggtgg ccgcactgat ggccatcggg
     7621 gatgagctgc gccggctgcg tctggctgtc agggaacagg gggcgcggga tgatagttaa
     7681 atttcatgcc aggggaaaag gtggtggcag tggtccggtt gattacctgc tggggaggga
     7741 gcgtaaccgc gaaggcgcaa cggtgcttca gggtaatccg gaagaagtcc gggaactcat
     7801 cgatgccacg ccatttgcga agaaatacac gtccggtgtt ctgtcgttcg cggagaagga
     7861 gctgccgccg ggaggacgtg aaaaagtgat ggcgagcttt gagcgtgtac tgatgcccgg
     7921 tctcgaaaaa aatcagtaca gcatcctgtg ggtggagcac caggacaagg gacggcttga
     7981 gctgaatttt gtcattccga acatggagct acagaccgga aaacgcctcc agccgtacta
     8041 cgaccgcgca gacaggccta gaattgatgc ctggcagacg ctggtaaatc accattacgg
     8101 gctgcatgac ccgaacgccc cggagaaccg caggacgctg acactccctg ataacctgcc
     8161 tgaaacgaaa caggcgctat ttcttccaga attgccatga ttttttcccc acgggaggcg
     8221 tcactggctc ccgtgttgtc ggcagctttg attcgataag cagcatcgcc tgtttcaggc
     8281 tgtctatgtg tgactgttga gctgtaacaa gttgtctcag gtgttcaatt tcatgttcta
     8341 gttgctttgt tttactggtt tcacctgttc tattaggtgt tacatgctgt tcatctgtta
     8401 cattgtcgat ctgttcatgg tgaacagctt taaatgcacc aaaaactcgt aaaagctctg
     8461 atgtatctat cttttttaca ccgttttcat ctgtgcatat ggacagtttt ccctttgata
     8521 tgtaacggtg aacagttgtt ctacttttgt ttgttagtct tgatgcttca ctgatagata
     8581 caagagccat aagaacctca gatccttccg tatttagcca gtatgttctc tagtgtggtt
     8641 cgttgttttt gcgtgagcca tgagaacgaa ccattgagat catacttact ttgcatgtca
     8701 ctcaaaaatt ttgcctcaaa actggtgagc tgaatttttg cagttaaagc atcgtgtagt
     8761 gtttttctta gtccgttatg taggtaggaa tctgatgtaa tggttgttgg tattttgtca
     8821 ccattcattt ttatctggtt gttctcaagt tcggttacga gatccatttg tctatctagt
     8881 tcaacttgga aaatcaacgt atcagtcggg cggcctcgct tatcaaccac caatttcata
     8941 ttgctgtaag tgtttaaatc tttacttatt ggtttcaaaa cccattggtt aagcctttta
     9001 aactcatggt agttattttc aagcattaac atgaacttaa attcatcaag gctaatctct
     9061 atatttgcct tgtgagtttt cttttgtgtt agttctttta ataaccactc ataaatcctc
     9121 atagagtatt tgttttcaaa agacttaaca tgttccagat tatattttat gaattttttt
     9181 aactggaaaa gataaggcaa tatctcttca ctaaaaacta attctaattt ttcgcttgag
     9241 aacttggcat agtttgtcca ctggaaaatc tcaaagcctt taaccaaagg attcctgatt
     9301 tccacagttc tcgtcatcag ctctctggtt gctttagcta atacaccata agcattttcc
     9361 ctactgatgt tcatcatctg aacgtattgg ttataagtga acgataccgt ccgttctttc
     9421 cttgtagggt tttcaatcgt ggggttgagt agtgccacac agcataaaat tagcttggtt
     9481 tcatgctccg ttaagtcata gcgactaatc gctagttcat ttgctttgaa aacaactaat
     9541 tcagacatac atctcaattg gtctaggtga ttttaatcac tataccaatt gagatgggct
     9601 agtcaatgat aattactagt ccttttcctt tgagttgtgg gtatctgtaa attctgctag
     9661 acctttgctg gaaaacttgt aaattctgct agaccctctg taaattccgc tagacctttg
     9721 tgtgtttttt ttgtttatat tcaagtggtt ataatttata gaataaagaa agaataaaaa
     9781 aagataaaaa gaatagatcc cagccctgtg tataactcac tactttagtc agttccgcag
     9841 tattacaaaa ggatgtcgca aacgctgttt gctcctctac aaaacagacc ttaaaaccct
     9901 aaaggcttaa gtagcaccct cgcaagctcg ggcaaatcgc tgaatattcc ttttgtctcc
     9961 gaccatcagg cacctgagtc gctgtctttt tcgtgacatt cagttcgctg cgctcacggc
    10021 tctggcagtg aatgggggta aatggcacta caggcgccca acagtccccc ggccacgggg
    10081 cctgccacca tacccacgcc gaaacaagcg ctgatgtccg gcggtgcttt tgccgttacg
    10141 caccaccccg tcagtagctg aacaggaggg acagctgata gaaacagaag ccactggagc
    10201 acctcaaaaa caccatcata cactaaatca gtaagttggc agcatcaccc gacgcacttt
    10261 gcgccgaata aatacctgtg acggaagatc acttcgcaga ataaataaat cctggtgtcc
    10321 ctgttgatac cgggaagccc tgggccaact tttggcgaaa atgagacgtt gatcggcacg
    10381 taagaggttc caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt
    10441 tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca
    10501 ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc
    10561 aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga
    10621 aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc
    10681 atccggaatt c
//

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