pcDNA3.1-CMV-CFP;UBC-Cre25nt

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载体基本信息

质粒名称: pcDNA3.1-CMV-CFP;UBC-Cre25nt
出品公司: Addgene
目录编号: 65727
存储实验室: Jacco van Rheenen

载体骨架基本信息
载体名称: pcDNA3.1
空载体大小: 8225bp
完整质粒大小: 8228 bp
载体类型: 哺乳细胞表达、Cre/Lox
筛选标记: Zeocin
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: CMV promoter driving CFP expression
启动子: CMV
克隆方法: 限制性内切酶
5’克隆位点: Mlul(未破坏)
3’克隆位点: EcoRI(未破坏)

文献引用方法
材料和方法部分: pcDNA3.1-CMV-CFP;UBC-Cre25nt was a gift from Jacco van Rheenen (Addgene plasmid # 65727 ; http://n2t.net/addgene:65727 ; RRID:Addgene_65727)
参考文献部分: In Vivo imaging reveals extracellular vesicle-mediated phenocopying of metastatic behavior. Zomer A, Maynard C, Verweij FJ, Kamermans A, Schafer R, Beerling E, Schiffelers RM, de Wit E, Berenguer J, Ellenbroek SI, Wurdinger T, Pegtel DM, van Rheenen J. Cell. 2015 May 21;161(5):1046-57. doi: 10.1016/j.cell.2015.04.042. 10.1016/j.cell.2015.04.042 PubMed 26000481

载体图谱质粒图谱和多克隆位点信息

pcDNA3.1-CMV-CFP;UBC-Cre25nt载体图谱

载体简介

载体序列

LOCUS       pcDNA3.1-CMV-CFP;UBC-Cre25nt                8228 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses Cre recombinase with a 25 nucleotide extracellular vesicle
            targeting sequence under the UBC promotor and CFP under the CMV 
            promotor..
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.1-CMV-CFP;UBC-Cre25nt
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8228)
  AUTHORS   Zomer A, Maynard C, Verweij FJ, Kamermans A, Schafer R, Beerling E, 
            Schiffelers RM, de Wit E, Berenguer J, Ellenbroek SI, Wurdinger T, 
            Pegtel DM, van Rheenen J
  TITLE     In Vivo imaging reveals extracellular vesicle-mediated phenocopying 
            of metastatic behavior.
  JOURNAL   Cell. 2015 May 21;161(5):1046-57. doi: 10.1016/j.cell.2015.04.042.
  PUBMED    26000481
REFERENCE   2  (bases 1 to 8228)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8228
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..440
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        441..644
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     595..615
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     689..708
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        689..707
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      730..739
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             736..1455
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(781..802)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1042..1061)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1389..1410
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1465..1689
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        1728..2939
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2776..2795
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             2962..3993
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     primer_bind     complement(4062..4079)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4068..4292
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4338..4766
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4425..4444)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4635..4656
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4775..4795)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4780..5109
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4960..5095
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5022..5041
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5157..5204
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5223..5597
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5727..5848
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5764..5783)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5818..5837
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5897..5913)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5897..5913)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5910..5932)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5921..5937
                     /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(5945..5975)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5990..6011
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6128..6145)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6299..6887)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6379..6398)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7058..7918)
                     /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     7681..7700
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7919..8023)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8098..8117)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
       61 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      121 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      181 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      241 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      301 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      361 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      421 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      481 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      541 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      601 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      661 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      721 gctagctttg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      781 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      841 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
      901 gtgccctggc ccaccctcgt gaccaccctg acctggggcg tgcagtgctt cagccgctac
      961 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1021 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1081 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1141 aacatcctgg ggcacaagct ggagtacaac tacatcagcc acaacgtcta tatcaccgcc
     1201 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     1261 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1321 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     1381 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1441 gagctgtaca agtaaggatc ccgactgtgc cttctagttg ccagccatct gttgtttgcc
     1501 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     1561 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     1621 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     1681 gctctatggg atctttttga attcgccctt ggccaaggca gccctggggc ctccgcgccg
     1741 ggttttggcg cctcccgcgg gcgcccccct cctcacggcg agcgctgcca cgtcagacga
     1801 agggcgcagc gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc
     1861 ataagactcg gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg
     1921 tgactctagg gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct
     1981 tctcggcgat tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga
     2041 cgcgccgggt gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt
     2101 tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg ctgggctggc cggggctttc
     2161 gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa gggctgtagt
     2221 ctgggtccgc gagcaaggtt gccctgaact gggggttggg gggagcgcag caaaatggcg
     2281 gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca
     2341 aggtgggggg catggtgggc ggcaagaacc caaggtcttg aggccttcgc taatgcggga
     2401 aagctcttat tcgggtgaga tgggctgggg caccatctgg ggaccctgac gtgaagtttg
     2461 tcactgactg gagaactcgg tttgtcgtct gttgcggggg cggcagttat ggcggtgccg
     2521 ttgggcagtg cacccgtacc tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg
     2581 ttctgttggc ttataatgca gggtggggcc acctgccggt aggtgtgcgg taggcttttc
     2641 tccgtcgcag gacgcagggt tcgggcctag ggtaggctct cctgaatcga caggcgccgg
     2701 acctctggtg aggggaggga taagtgaggc gtcagtttct ttggtcggtt ttatgtacct
     2761 atcttcttaa gtagctgaag ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt
     2821 tttgtgaagt tttttaggca ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc
     2881 agtgttagac tagtaaattg tccgctaaat tctggccgtt tttggctttt ttgttagacg
     2941 aagcttgggg tacccgccac catgtccaat ttactgaccg tacaccaaaa tttgcctgca
     3001 ttaccggtcg atgcaacgag tgatgaggtt cgcaagaacc tgatggacat gttcagggat
     3061 cgccaggcgt tttctgagca tacctggaaa atgcttctgt ccgtttgccg gtcgtgggcg
     3121 gcatggtgca agttgaataa ccggaaatgg tttcccgcag aacctgaaga tgttcgcgat
     3181 tatcttctat atcttcaggc gcgcggtctg gcagtaaaaa ctatccagca acatttgggc
     3241 cagctaaaca tgcttcatcg tcggtccggg ctgccacgac caagtgacag caatgctgtt
     3301 tcactggtta tgcggcggat ccgaaaagaa aacgttgatg ccggtgaacg tgcaaaacag
     3361 gctctagcgt tcgaacgcac tgatttcgac caggttcgtt cactcatgga aaatagcgat
     3421 cgctgccagg atatacgtaa tctggcattt ctggggattg cttataacac cctgttacgt
     3481 atagccgaaa ttgccaggat cagggttaaa gatatctcac gtactgacgg tgggagaatg
     3541 ttaatccata ttggcagaac gaaaacgctg gttagcaccg caggtgtaga gaaggcactt
     3601 agcctggggg taactaaact ggtcgagcga tggatttccg tctctggtgt agctgatgat
     3661 ccgaataact acctgttttg ccgggtcaga aaaaatggtg ttgccgcgcc atctgccacc
     3721 agccagctat caactcgcgc cctggaaggg atttttgaag caactcatcg attgatttac
     3781 ggcgctaagg atgactctgg tcagagatac ctggcctggt ctggacacag tgcccgtgtc
     3841 ggagccgcgc gagatatggc ccgcgctgga gtttcaatac cggagatcat gcaagctggt
     3901 ggctggacca atgtaaatat tgtcatgaac tatatccgta acctggatag tgaaacaggg
     3961 gcaatggtgc gcctgctgga agatggcgat taggaatcca ccctgccgcc tggactccgc
     4021 ctgtgaatcc tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     4081 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     4141 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     4201 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     4261 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     4321 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4381 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4441 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4501 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4561 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4621 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4681 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4741 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4801 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4861 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4921 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4981 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     5041 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     5101 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg
     5161 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     5221 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
     5281 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
     5341 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
     5401 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
     5461 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
     5521 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
     5581 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
     5641 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5701 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5761 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5821 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5881 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5941 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     6001 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     6061 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6121 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6181 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6241 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6301 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6361 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6421 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6481 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6541 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6601 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6661 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6721 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6781 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6841 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6901 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6961 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     7021 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7081 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7141 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7201 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7261 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7321 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7381 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7441 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7501 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7561 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7621 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7681 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7741 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7801 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7861 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7921 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7981 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     8041 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     8101 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     8161 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     8221 catgaaga
//

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