pcDNA-GalNAc-T2-GFP-ESCargo(FTV)

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载体基本信息

质粒名称: pcDNA-GalNAc-T2-GFP-ESCargo(FTV)
出品公司: Addgene
目录编号: 140163
存储实验室: Benjamin Glick

载体骨架基本信息
载体名称: pcDNA5/FRT/TO
载体出品公司: --
空载体大小: --
完整质粒大小: --
载体修饰: --
载体类型: 哺乳细胞表达
筛选标记: --
载体抗性: 氨苄霉素
生长温度: 37 ℃
宿主菌: DH5a
拷贝数: --

插入基因信息
插入基因名称: GalNAc-T2-msGFP2 and piGH-FTVNTT-DsRed-Express2-FKBP(LV)(C22V)
物种来源: H. sapiens (human), Synthetic
插入大小: --
基因突变: --
GenBank编号 --
启动子: EF-1a
标签/融合蛋白: ER信号序列(MGWSCIILFLVATATGAHS)(主干上的N端)(插入上的N端子)
克隆方法: Gibson克隆
5’克隆位点: --
3’克隆位点: --
5’测序引物 --
3’测序引物 --

文献引用方法
材料和方法部分: pcDNA-GalNAc-T2-GFP-ESCargo(FTV) was a gift from Benjamin Glick (Addgene plasmid # 140163 ; http://n2t.net/addgene:140163 ; RRID:Addgene_140163)
参考文献部分: ESCargo: a regulatable fluorescent secretory cargo for diverse model organisms. Casler JC, Zajac AL, Valbuena FM, Sparvoli D, Jeyifous O, Turkewitz AP, Horne-Badovinac S, Green WN, Glick BS. Mol Biol Cell. 2020 Oct 28:mbcE20090591. doi: 10.1091/mbc.E20-09-0591. 10.1091/mbc.E20-09-0591 PubMed 33112725

载体图谱质粒图谱和多克隆位点信息

pcDNA-GalNAc-T2-GFP-ESCargo(FTV)载体图谱

载体简介

载体序列

LOCUS       pcDNA-GalNAc-T2-GFP-ESCargo(FTV)         10051 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses a regulatable secretory cargo for mammalian cells and a 
            Golgi marker.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA-GalNAc-T2-GFP-ESCargo(FTV)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10051)
  AUTHORS   Casler JC, Zajac AL, Valbuena FM, Sparvoli D, Jeyifous O, Turkewitz 
            AP, Horne-Badovinac S, Green WN, Glick BS
  TITLE     ESCargo: a regulatable fluorescent secretory cargo for diverse model
            organisms.
  JOURNAL   Mol Biol Cell. 2020 Oct 28:mbcE20090591. doi: 
            10.1091/mbc.E20-09-0591.
  PUBMED    33112725
REFERENCE   2  (bases 1 to 10051)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10051
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        51..1213
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          282..1204
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1161..1181
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2964..3671
                     /codon_start=1
                     /product="photostable monomeric superfolder GFP (Valbuena 
                     et al., 2020)"
                     /label=msGFP2
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     primer_bind     complement(3006..3027)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     3614..3635
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3701..3925
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        4019..4398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4399..4602
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4553..4573
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4612..4636
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4727..4736
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     sig_peptide     join(4733..4777,4860..4871)
                     /product="modified signal sequence from the mouse 
                     immunoglobulin heavy chain variable region"
                     /label=IgH signal sequence
     primer_bind     complement(5067..5087)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     5285..5304
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5479..5502
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             5574..5894
                     /codon_start=1
                     /product="destabilization domain that can be stabilized by 
                     Shield1 in the ProteoTuner(TM) system"
                     /label=DD
                     /note="L106P mutant of FKBP12"
                     /translation="GVQVETISPGDGRTFPKRGQTVVVHYTGMLEDGKKLDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGVIPPHATLVFDVELL
                     KLE"
     polyA_signal    6028..6252
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6385..6404)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    6536..6583
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             6591..7613
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    7743..7864
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7780..7799)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7834..7853
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7913..7929)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7913..7929)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7926..7948)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7937..7953
                     /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(7961..7991)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8006..8027
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8144..8161)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8315..8903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8395..8414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9074..9934)
                     /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     9697..9716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9935..10039)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaagtgcca cctgacgtcg acggatcggg agatccttca agcttcgtga ggctccggtg
       61 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      121 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      181 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      241 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      301 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      361 ggctccagta cgtgattctt gatcccgagc tggagccagg ggcgggcctt gcgctttagg
      421 agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg
      481 aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa
      541 tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca
      601 ggatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc
      661 ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg
      721 gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc
      781 gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct
      841 tcccggccct gctccagggg gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg
      901 tgagtcaccc acacaaagga aaggggcctt tccgtcctca gccgtcgctt catgtgactc
      961 cacggagtac cgggcgccgt ccaggcacct cgattagttc tggagctttt ggagtacgtc
     1021 gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga
     1081 gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga
     1141 gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt
     1201 tcaggtgtcg tgaacacgtg gccaccgcca ccatgaggcg gcgctcgagg atgctgctct
     1261 gcttcgcctt cctgtgggtg ctgggaatcg cctactacat gtactctgga ggaggatctg
     1321 cgctggccgg aggagcggga ggcggagccg gaaggaagga ggactggaat gaaattgacc
     1381 ccattaaaaa gaaagacctt catcacagca atggagaaga gaaagcacaa agcatggaga
     1441 ccctccctcc agggaaagta cggtggccag actttaacca ggaagcttat gttggaggga
     1501 cgatggtccg ctccgggcag gacccttacg cccgcaacaa gttcaaccag gtggagagtg
     1561 ataagcttcg aatggacaga gccatccctg acacccggca tgaccagtgt cagcggaagc
     1621 agtggcgggt ggatctgccg gccaccagcg tggtgatcac gtttcacaat gaagccaggt
     1681 cggccctact caggaccgtg gtcagcgtgc ttaagaaaag cccgccccat ctcataaaag
     1741 aaatcatctt ggtggatgac tacagcaatg atcctgagga cggggctctc ttggggaaaa
     1801 ttgagaaagt gcgagttctt agaaatgatc gacgagaagg cctcatgcgc tcacgggttc
     1861 ggggggccga tgctgcccaa gccaaggtcc tgaccttcct ggacagtcac tgcgagtgta
     1921 atgagcactg gctggagccc ctcctggaaa gggtggcgga ggacaggact cgggttgtgt
     1981 cacccatcat cgatgtcatt aatatggaca actttcagta tgtgggggca tctgctgact
     2041 tgaagggcgg ttttgattgg aacttggtat tcaagtggga ttacatgacg cctgagcaga
     2101 gaaggtcccg gcaggggaac ccagtcgccc ctataaaaac ccccatgatt gctggtgggc
     2161 tgtttgtgat ggataagttc tattttgaag aactggggaa gtacgacatg atgatggatg
     2221 tgtggggagg agagaaccta gagatctcgt tccgcgtgtg gcagtgtggt ggcagcctgg
     2281 agatcatccc gtgcagccgt gtgggacacg tgttccggaa gcagcacccc tacacgttcc
     2341 cgggtggcag tggcactgtc tttgcccgaa acacccgccg ggcagcagag gtctggatgg
     2401 atgaatacaa aaatttctat tatgcagcag tgccttctgc tagaaacgtt ccttatggaa
     2461 atattcagag cagattggag cttaggaaga aactcagctg caagcctttc aaatggtacc
     2521 ttgaaaatgt ctatccagag ttaagggttc cagaccatca ggatatagct tttggggcct
     2581 tgcagcaggg aactaactgc ctcgacactt tgggacactt tgctgatggt gtggttggag
     2641 tttatgaatg tcacaatgct gggggaaacc aggaatgggc cttgacgaag gagaagtcgg
     2701 tgaagcacat ggatttgtgc cttactgtgg tggaccgggc accgggctct cttataaagc
     2761 tgcagggctg ccgagaaaat gacagcagac agaaatggga acagatcgag ggcaactcca
     2821 agctgaggca cgtgggcagc aacctgtgcc tggacagtcg cacggccaag agcgggggcc
     2881 taagcgtgga ggtgtgtggc ccggcccttt cgcagcagtg gaagttcacg ctcaacctgc
     2941 agcagggagg aagcggagga agcatggata gcactgagag cctgttcacc ggggtggtgc
     3001 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg cgcggcgagg
     3061 gcgagggcga tgccaccaac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     3121 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     3181 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     3241 tccaggagcg caccatcacc ttcaaggacg acggcaccta caagacccgc gccgaggtga
     3301 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     3361 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     3421 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacgtggagg
     3481 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     3541 tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg
     3601 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actggaggca
     3661 gtggcggttc ttagtctaga agctcgctga tcagcctcga ctgtgccttc tagttgccag
     3721 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3781 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3841 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3901 gctggggatg gcccgggctc tatggcttct gaggcggaaa gaacgaattg catgaagaat
     3961 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
     4021 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     4081 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     4141 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     4201 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     4261 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     4321 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     4381 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     4441 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     4501 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     4561 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag cttcgtcgac gagctcgttt
     4621 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca
     4681 ccgggaccga tccagcctcc ggactctagc gtttaaactt aagcttgcca ccatgggatg
     4741 gagctgtatc atcctcttct tggtagcaac agctacaggt aaggggttaa cagtagcagg
     4801 cttgaggtct ggacatatat atgggtgaca atgacatcca ctttgccttt ctctccacag
     4861 gcgcgcactc ctttaccgtg aacaccaccg atagcactga gaacgtcatc aagcccttca
     4921 tgcgcttcaa ggtgcacatg gagggctcag taaatggtca tgagttcgag atcgagggcg
     4981 agggcgaggg caagccctac gagggcaccc agaccgccaa gctgcaggtg accaagggcg
     5041 gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc tccaaggtgt
     5101 acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc gagggcttca
     5161 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
     5221 ccctgcagga cggcaccttc atctaccacg tgaagttcat cggcgtgaac ttcccctccg
     5281 acggccccgt aatgcagaag aagactctgg gctgggagcc ctccaccgag cgcctgtacc
     5341 cccgcgacgg cgtgctgaag ggcgagatcc acaaggcgct gaagctgaag ggcggcggcc
     5401 actacctggt ggagttcaag tcaatctaca tggccaagaa gcccgtgaag ctgcccggct
     5461 actactacgt ggactccaag ctggacatca cctcccacaa cgaggactac accgtggtgg
     5521 agcagtacga gcgcgccgag gcccgccacc acctgttcca gggaggatcc agaggagtgc
     5581 aggtggaaac catctcccca ggagacgggc gcaccttccc caagcgcggc cagaccgttg
     5641 tggtgcacta caccgggatg cttgaagatg gaaagaaact agattcctcc cgggacagaa
     5701 acaagccctt taagtttatg ctaggcaagc aggaggtgat ccgaggctgg gaagaagggg
     5761 ttgcccagat gagtgtgggt cagagagcca aactgactat atctccagat tatgcctatg
     5821 gtgccactgg gcacccaggc gttatcccac cacatgccac tctcgtcttc gatgtggagc
     5881 ttctaaaact ggaatagtct agaagctcgc tgatcaggta ccgagctcgg atccactagt
     5941 ccagtgtggt ggaattctgc agatatccag cacagtggcg gccgctcgag tctagagggc
     6001 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     6061 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     6121 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     6181 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     6241 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     6301 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     6361 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     6421 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     6481 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt acctagaagt
     6541 tcctattccg aagttcctat tctctagaaa gtataggaac ttccttggcc aaaaagcctg
     6601 aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc
     6661 tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg
     6721 gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc
     6781 ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg
     6841 agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg
     6901 aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg
     6961 ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca
     7021 ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg
     7081 tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg
     7141 ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc
     7201 tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt
     7261 cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc
     7321 agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt
     7381 atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg
     7441 atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg
     7501 ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac
     7561 tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa tagcacgtac
     7621 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     7681 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     7741 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     7801 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     7861 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     7921 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     7981 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     8041 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     8101 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     8161 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     8221 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     8281 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     8341 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     8401 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     8461 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     8521 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     8581 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     8641 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     8701 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     8761 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     8821 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     8881 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     8941 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     9001 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     9061 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     9121 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     9181 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     9241 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     9301 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     9361 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     9421 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     9481 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     9541 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     9601 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     9661 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     9721 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     9781 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     9841 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     9901 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     9961 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
    10021 aacaaatagg ggttccgcgc acatttcccc g
//

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