vYM112_PB-pEF1a-optiPIN2-HA(Hygro)

价格:1200元

联系方式:I47-825O-882O

相关技术服务:质粒构建    基因合成    质粒大提

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载体基本信息

质粒名称: vYM112_PB-pEF1a-optiPIN2-HA(Hygro)
出品公司: Addgene
目录编号: 160045
存储实验室: Michael Elowitz

载体骨架基本信息
载体名称: PiggyBac
载体出品公司: SYSTEM BIOSCIENCES
载体类型: 哺乳动物表达,合成生物学
筛选标记: 潮霉素
载体抗性: 氨苄霉素,100 μg/mL
生长温度: 30 ℃
宿主菌: DH5a
拷贝数: 高拷贝

插入基因信息
插入基因名称: PIN2
物种来源: A. thaliana (mustard weed)
插入大小: 1941 bp
基因突变: codon optimized
GenBank编号 AF086907.1
启动子: pEF1s
标签/融合蛋白: HA(插入件上的C端子)
克隆方法: Gibson克隆
5’测序引物 taaggtagcgaaggtctgCGTAATG
3’测序引物 agggataacagggtaatgaggtTTAtgcgtagtctggtacgtcgtacggatagccgc

文献引用方法
材料和方法部分: vYM112_PB-pEF1a-optiPIN2-HA(Hygro) was a gift from Michael Elowitz (Addgene plasmid # 160045 ; http://n2t.net/addgene:160045 ; RRID:Addgene_160045)
参考文献部分: Synthetic mammalian signaling circuits for robust cell population control. Ma Y, Budde MW, Mayalu MN, Zhu J, Lu AC, Murray RM, Elowitz MB. Cell. 2022 Mar 17;185(6):967-979.e12. doi: 10.1016/j.cell.2022.01.026. Epub 2022 Mar 1. 10.1016/j.cell.2022.01.026 PubMed 35235768

载体图谱质粒图谱和多克隆位点信息

vYM112_PB-pEF1a-optiPIN2-HA(Hygro)载体图谱

载体简介

载体序列

LOCUS       vYM112_PB-pEF1a-optiPIN2-HA(Hygro)               9590 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  For expressing the active auxin pump PIN2.
ACCESSION   .
VERSION     .
KEYWORDS    vYM112_PB-pEF1a-optiPIN2-HA(Hygro)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9590)
  TITLE     Synthetic mammalian signaling circuits for robust cell population 
            control
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9590)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9590
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(59..76)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(230..818)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(310..329)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(989..1849)
                     /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     1612..1631
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1850..1954)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1980..2435)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2117..2138)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2329..2348
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2562..2584
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2576..2593
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2577..2593
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   2723..2785
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        3228..4406
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3459..4397
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4354..4374
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             6418..6444
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(6472..6489)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6478..6685
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6718..6738)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6723..7052
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6903..7038
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6965..6984
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             7101..8126
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     complement(8186..8206)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    8249..8370
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8286..8305)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8340..8359
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   8893..8927
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9418..9434)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9418..9434)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9431..9453)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9442..9458
                     /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(9466..9496)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9511..9532
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
       61 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      121 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      181 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
      241 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
      301 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
      361 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
      421 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
      481 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
      541 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
      601 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
      661 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
      721 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
      781 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
      841 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
      901 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
      961 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     1021 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1081 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gatccacgct
     1141 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1201 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     1261 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     1321 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     1381 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1441 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     1501 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     1561 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     1621 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     1681 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     1741 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     1801 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     1861 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     1921 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     1981 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2041 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2101 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2161 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     2221 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2281 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2341 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2401 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     2461 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     2521 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     2581 aacgacggcc agtgagcgcg cctcgttcat tcacgttttt gaacccgtgg aggacgggca
     2641 gactcgcggt gcaaatgtgt tttacagcgt gatggagcag atgaagatgc tcgacacgct
     2701 gcagaacacg cagctagatt aaccctagaa agataatcat attgtgacgt acgttaaaga
     2761 taatcatgtg taaaattgac gcatgtgttt tatcggtctg tatatcgagg tttatttatt
     2821 aatttgaata gatattaagt tttattatat ttacacttac atactaataa taaattcaac
     2881 aaacaattta tttatgttta tttatttatt aaaaaaaaca aaaactcaaa atttcttcta
     2941 taaagtaaca aaacttttat gagggacagc ccccccccaa agcccccagg gatgtaatta
     3001 cgtccctccc ccgctagggg gcagcagcga gccgcccggg gctccgctcc ggtccggcgc
     3061 tccccccgca tccccgagcc ggcagcgtgc ggggacagcc cgggcacggg gaaggtggca
     3121 cgggatcgct ttcctctgaa cgcttctcgc tgctctttga gcctgcagac acctgggggg
     3181 atacggggaa aaggcctcca cggccagcac ctgcgtgacg tacgtgaggc tccggtgccc
     3241 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     3301 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     3361 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     3421 acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc
     3481 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc
     3541 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
     3601 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
     3661 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
     3721 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
     3781 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
     3841 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
     3901 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
     3961 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
     4021 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg
     4081 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
     4141 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
     4201 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     4261 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     4321 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     4381 tttttcttcc atttcaggtg tcgtgaacct tcacgcaggt ggaattctaa ctataacggt
     4441 cctaaggtag cgaaggtctg cgtaatgatc actggcaaag atatgtacga tgtccttgct
     4501 gcaatggtac cactctatgt agcaatgatt cttgcttatg gcagtgtccg ctggtggggc
     4561 atatttacac ccgatcagtg cagcgggatc aatcggttcg ttgccgtttt tgcagtaccc
     4621 ctcctttcat tccacttcat ttcaagcaat gatccttacg caatgaatta tcactttctc
     4681 gctgcagaca gtcttcagaa agtggtaatc ttggcagcac tctttctttg gcaggccttc
     4741 tctagaagag gctctttgga gtggatgatt accctcttct ctctttctac tctccccaac
     4801 acattggtga tgggcatacc tcttcttaga gcaatgtacg gggatttctc tggcaacctc
     4861 atggtccaga tcgtggtgct tcagtctatc atatggtata cccttatgct gttccttttt
     4921 gagttccgcg gggcaaaact gctcataagt gaacaattcc cagagaccgc cgggtctatc
     4981 accagcttcc gggtggactc tgacgtaata agcctgaacg ggagagaacc ccttcaaaca
     5041 gacgctgaaa taggagacga tggtaagttg catgtcgtag tacggaggtc aagcgctgct
     5101 tcctcaatga tatctagttt caacaaatcc catggaggcg gccttaattc tagtatgata
     5161 acaccaagag ccagcaatct cacaggcgta gaaatttact cagtgcagtc aagtagggaa
     5221 ccaaccccac gcgcctcctc ttttaatcaa actgactttt atgccatgtt taacgcaagt
     5281 aaggccccta gccctagaca cggctatact aatagttacg ggggcgctgg ggctggccca
     5341 ggaggtgatg tctacagttt gcaatcaagt aagggggtaa ctcctcgcac ttcaaacttt
     5401 gacgaggaag taatgaagac tgccaagaaa gcaggaaggg gcggacgcag tatgtcaggg
     5461 gaactgtaca acaataactc agttccttct tatccaccac ctaacccaat gtttacaggc
     5521 tcaacctctg gtgcatcagg cgtaaagaaa aaggaaagcg gtggaggcgg ctccggaggt
     5581 ggcgtcggcg taggaggcca gaacaaagag atgaacatgt ttgtttggag ttccagtgct
     5641 agcccagtca gtgaggctaa tgccaaaaac gcaatgaccc ggggttctag cacagatgtt
     5701 tctaccgacc caaaggtgtc tatccctcca catgataatt tggcaaccaa agccatgcaa
     5761 aacctcatcg aaaatatgag tcctggcaga aagggccatg tggagatgga tcaagatgga
     5821 aacaatggtg gtaaaagccc ttatatgggg aaaaaaggct cagacgtcga agatggcgga
     5881 cctggaccta gaaaacagca gatgccccca gcttccgtaa tgacacgact catccttatt
     5941 atggtgtggc ggaaactcat acgcaatcca aacacctaca gcagcttgtt tgggctcgcc
     6001 tggtctcttg tatcattcaa gtggaacatt aaaatgccaa caataatgag cgggtctatc
     6061 tctattcttt cagatgctgg tttgggtatg gccatgttta gtttgggcct gttcatggct
     6121 ctccaaccca agataattgc ttgtggaaag tctgtagccg gctttgctat ggccgtgcgg
     6181 ttcctcacag gacctgccgt aatagcagct acttcaatcg ccataggtat caggggtgac
     6241 ctgcttcaca ttgctattgt tcaagccgcc ttgcctcagg gcatcgtacc tttcgtcttt
     6301 gctaaggaat ataacgttca tcctgacata ctctccaccg ctgtcatctt cgggatgttg
     6361 gttgcattgc cagtaacagt tctgtactac gttcttcttg gccttggcgg aagcggctat
     6421 ccgtacgacg taccagacta cgcataaacc tcattaccct gttatcccta gcctcgactg
     6481 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     6541 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     6601 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     6661 aagacaatag caggcatgct ggggacgcgg ccgcaattta acgcgaatta attctgtgga
     6721 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     6781 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     6841 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     6901 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     6961 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     7021 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     7081 tcggatctga tcagcacgtg atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt
     7141 ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat
     7201 ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg
     7261 ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga
     7321 ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc
     7381 gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc
     7441 cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg
     7501 gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga
     7561 ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg
     7621 tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc
     7681 tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg
     7741 tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct
     7801 tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc
     7861 cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac
     7921 tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg
     7981 acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg
     8041 cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca
     8101 gcactcgtcc gagggcaaag gaataggtcg acaatcaacc tctggattac aaaatttgtg
     8161 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     8221 taatgccttt gtatcatccc tagaattcaa cttgtttatt gcagcttata atggttacaa
     8281 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     8341 tggtttgtcc aaactcatca atgtatctta ggcccattgg tatggctttt tccccgtatc
     8401 cccccaggtg tctgcaggct caaagagcag cgagaagcgt tcagaggaaa gcgatcccgt
     8461 gccaccttcc ccgtgcccgg gctgtccccg cacgctgccg gctcggggat gcggggggag
     8521 cgccggaccg gagcggagcc ccgggcggct cgctgctgcc ccctagcggg ggagggacgt
     8581 aattacatcc ctgggggctt tggggggggg ctgtccctga tatctataac aagaaaatat
     8641 atatataata agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt
     8701 taaatcttaa aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat
     8761 agttcaaaat cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg
     8821 cgactgagat gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat
     8881 atttcaagaa tgcatgcgtc aattttacgc agactatctt tctagggtta atctagctgc
     8941 atcaggatca tatcgtcggg tcttttttcc ggctcagtca tcgcccaagc tggcgctatc
     9001 tgggcatcgg ggaggaagaa gcccgtgcct tttcccgcga ggttgaagcg gcatggaaag
     9061 agtttgccga ggatgactgc tgctgcattg acgttgagcg aaaacgcacg tttaccatga
     9121 tgattcggga aggtgtggcc atgcacgcct ttaacggtga actgttcgtt caggccacct
     9181 gggataccag ttcgtcgcgg cttttccgga cacagttccg gatggtcagc ccgaagcgca
     9241 tcagcaaccc gaacaatacc ggcgacagcc ggaactgccg tgccggtgtg cagattaatg
     9301 acagcggtgc ggcgctggga tattacgtca gcgaggacgg gtatcctggc tggatgccgc
     9361 agaaatggac atggataccc cgtgagttac ccggcgggcg cgcttggcgt aatcatggtc
     9421 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     9481 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     9541 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
//

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