LOCUS       sequence-469472-       13853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Von Willebrand factor fused in tandem to superecliptic pHluorin and 
            inserted into pmRFP1-N1 vector from Addgene (Plasmid No: 
            54635)/Monitoring of WPB pH, Visualization of WPB exocytosis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13853)
  AUTHORS   Terglane J, Menche D, Gerke V
  TITLE     Acidification of endothelial Weibel-Palade bodies is mediated by the
            vacuolar-type H+-ATPase.
  JOURNAL   PLoS One. 2022 Jun 29;17(6):e0270299. doi: 
            10.1371/journal.pone.0270299. eCollection 2022.
  PUBMED    35767558
REFERENCE   2  (bases 1 to 13853)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2022-06-29"; volume: "17(6):e0270299. doi"; pages: " 
            10.1371/journal.pone.0270299. eCollection 2022"
COMMENT     Sequence Label: VWF-pHluorin-mRFP
FEATURES             Location/Qualifiers
     source          1..13853
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(35..490)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(172..193)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     384..403
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        517..621
                     /gene="bla"
                     /label=AmpR promoter
     promoter        623..980
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        623..814
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(646..666)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      831..966
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1015..1809
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1015..1809
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1069..1088)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1679..1698
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1997..2016)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2041..2088
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2417..3005
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2906..2925
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3180..3483
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3484..3687
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3638..3658
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5309..5320
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             12186..12896
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Ng et al., 2002)"
                     /label=superecliptic pHluorin
                     /note="excited strongly by blue light at neutral pH but 
                     minimally fluorescent at acidic pH"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(12213..12241)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     12836..12856
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     regulatory      12954..12963
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             12960..13637
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             12960..13637
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(13090..13108)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     13358..13377
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     13552..13575
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    join(13760..13853,1..28)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13797..13816)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     join(13851..13853,1..17)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
       61 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
      121 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
      181 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
      241 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
      301 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
      361 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
      421 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
      481 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
      541 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
      601 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
      661 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
      721 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
      781 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
      841 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
      901 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
      961 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     1021 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     1081 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     1141 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     1201 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     1261 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     1321 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     1381 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     1441 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     1501 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     1561 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     1621 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     1681 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     1741 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     1801 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     1861 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     1921 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     1981 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     2041 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     2101 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     2161 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     2221 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     2281 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     2341 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     2401 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     2461 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     2521 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     2581 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2641 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2701 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2761 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2821 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2881 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2941 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3001 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3061 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt
     3121 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     3181 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     3241 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     3301 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     3361 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     3421 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     3481 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     3541 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     3601 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     3661 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcctcga
     3721 gctcaagctt cggattcatg attcctgcca gatttgccgg ggtgctgctt gctctggccc
     3781 tcattttgcc agggaccctt tgtgcagaag gaactcgcgg caggtcatcc acggcccgat
     3841 gcagcctttt cggaagtgac ttcgtcaaca cctttgatgg gagcatgtac agctttgcgg
     3901 gatactgcag ttacctcctg gcagggggct gccagaaacg ctccttctcg attattgggg
     3961 acttccagaa tggcaagaga gtgagcctct ccgtgtatct tggggaattt tttgacatcc
     4021 atttgtttgt caatggtacc gtgacacagg gggaccaaag agtctccatg ccctatgcct
     4081 ccaaagggct gtatctagaa actgaggctg ggtactacaa gctgtccggt gaggcctatg
     4141 gctttgtggc caggatcgat ggcagcggca actttcaagt cctgctgtca gacagatact
     4201 tcaacaagac ctgcgggctg tgtggcaact ttaacatctt tgctgaagat gactttatga
     4261 cccaagaagg gaccttgacc tcggaccctt atgactttgc caactcatgg gctctgagca
     4321 gtggagaaca gtggtgtgaa cgggcatctc ctcccagcag ctcatgcaac atctcctctg
     4381 gggaaatgca gaagggcctg tgggagcagt gccagcttct gaagagcacc tcggtgtttg
     4441 cccgctgcca ccctctggtg gaccccgagc cttttgtggc cctgtgtgag aagactttgt
     4501 gtgagtgtgc tggggggctg gagtgcgcct gccctgccct cctggagtac gcccggacct
     4561 gtgcccagga gggaatggtg ctgtacggct ggaccgacca cagcgcgtgc agcccagtgt
     4621 gccctgctgg tatggagtat aggcagtgtg tgtccccttg cgccaggacc tgccagagcc
     4681 tgcacatcaa tgaaatgtgt caggagcgat gcgtggatgg ctgcagctgc cctgagggac
     4741 agctcctgga tgaaggcctc tgcgtggaga gcaccgagtg tccctgcgtg cattccggaa
     4801 agcgctaccc tcccggcacc tccctctctc gagactgcaa cacctgcatt tgccgaaaca
     4861 gccagtggat ctgcagcaat gaagaatgtc caggggagtg ccttgtcaca ggtcaatcac
     4921 acttcaagag ctttgacaac agatacttca ccttcagtgg gatctgccag tacctgctgg
     4981 cccgggattg ccaggaccac tccttctcca ttgtcattga gactgtccag tgtgctgatg
     5041 accgcgacgc tgtgtgcacc cgctccgtca ccgtccggct gcctggcctg cacaacagcc
     5101 ttgtgaaact gaagcatggg gcaggagttg ccatggatgg ccaggacgtc cagctccccc
     5161 tcctgaaagg tgacctccgc atccagcata cagtgacggc ctccgtgcgc ctcagctacg
     5221 gggaggacct gcagatggac tgggatggcc gcgggaggct gctggtgaag ctgtcccccg
     5281 tctatgccgg gaagacctgc ggcctgtgtg ggaattacaa tggcaaccag ggcgacgact
     5341 tccttacccc ctctgggctg gcggagcccc gggtggagga cttcgggaac gcctggaagc
     5401 tgcacgggga ctgccaggac ctgcagaagc agcacagcga tccctgcgcc ctcaacccgc
     5461 gcatgaccag gttctccgag gaggcgtgcg cggtcctgac gtcccccaca ttcgaggcct
     5521 gccatcgtgc cgtcagcccg ctgccctacc tgcggaactg ccgctacgac gtgtgctcct
     5581 gctcggacgg ccgcgagtgc ctgtgcggcg ccctggccag ctatgccgcg gcctgcgcgg
     5641 ggagaggcgt gcgcgtcgcg tggcgcgagc caggccgctg tgagctgaac tgcccgaaag
     5701 gccaggtgta cctgcagtgc gggaccccct gcaacctgac ctgccgctct ctctcttacc
     5761 cggatgagga atgcaatgag gcctgcctgg agggctgctt ctgcccccca gggctctaca
     5821 tggatgagag gggggactgc gtgcccaagg cccagtgccc ctgttactat gacggtgaga
     5881 tcttccagcc agaagacatc ttctcagacc atcacaccat gtgctactgt gaggatggct
     5941 tcatgcactg taccatgagt ggagtccccg gaagcttgct gcctgacgct gtcctcagca
     6001 gtcccctgtc tcatcgcagc aaaaggagcc tatcctgtcg gccccccatg gtcaagctgg
     6061 tgtgtcccgc tgacaacctg cgggctgaag ggctcgagtg taccaaaacg tgccagaact
     6121 atgacctgga gtgcatgagc atgggctgtg tctctggctg cctctgcccc ccgggcatgg
     6181 tccggcatga gaacagatgt gtggccctgg aaaggtgtcc ctgcttccat cagggcaagg
     6241 agtatgcccc tggagaaaca gtgaagattg gctgcaacac ttgtgtctgt cgggaccgga
     6301 agtggaactg cacagaccat gtgtgtgatg ccacgtgctc cacgatcggc atggcccact
     6361 acctcacctt cgacgggctc aaatacctgt tccccgggga gtgccagtac gttctggtgc
     6421 aggattactg cggcagtaac cctgggacct ttcggatcct agtggggaat aagggatgca
     6481 gccacccctc agtgaaatgc aagaaacggg tcaccatcct ggtggaggga ggagagattg
     6541 agctgtttga cggggaggtg aatgtgaaga ggcccatgaa ggatgagact cactttgagg
     6601 tggtggagtc tggccggtac atcattctgc tgctgggcaa agccctctcc gtggtctggg
     6661 accgccacct gagcatctcc gtggtcctga agcagacata ccaggagaaa gtgtgtggcc
     6721 tgtgtgggaa ttttgatggc atccagaaca atgacctcac cagcagcaac ctccaagtgg
     6781 aggaagaccc tgtggacttt gggaactcct ggaaagtgag ctcgcagtgt gctgacacca
     6841 gaaaagtgcc tctggactca tcccctgcca cctgccataa caacatcatg aagcagacga
     6901 tggtggattc ctcctgtaga atccttacca gtgacgtctt ccaggactgc aacaagctgg
     6961 tggaccccga gccatatctg gatgtctgca tttacgacac ctgctcctgt gagtccattg
     7021 gggactgcgc ctgcttctgc gacaccattg ctgcctatgc ccacgtgtgt gcccagcatg
     7081 gcaaggtggt gacctggagg acggccacat tgtgccccca gagctgcgag gagaggaatc
     7141 tccgggagaa cgggtatgag tgtgagtggc gctataacag ctgtgcacct gcctgtcaag
     7201 tcacgtgtca gcaccctgag ccactggcct gccctgtgca gtgtgtggag ggctgccatg
     7261 cccactgccc tccagggaaa atcctggatg agcttttgca gacctgcgtt gaccctgaag
     7321 actgtccagt gtgtgaggtg gctggccggc gttttgcctc aggaaagaaa gtcaccttga
     7381 atcccagtga ccctgagcac tgccagattt gccactgtga tgttgtcaac ctcacctgtg
     7441 aagcctgcca ggagccggga ggcctggtgg tgcctcccac agatgccccg gtgagcccca
     7501 ccactctgta tgtggaggac atctcggaac cgccgttgca cgatttctac tgcagcaggc
     7561 tactggacct ggtcttcctg ctggatggct cctccaggct gtccgaggct gagtttgaag
     7621 tgctgaaggc ctttgtggtg gacatgatgg agcggctgcg catctcccag aagtgggtcc
     7681 gcgtggccgt ggtggagtac cacgacggct cccacgccta catcgggctc aaggaccgga
     7741 agcgaccgtc agagctgcgg cgcattgcca gccaggtgaa gtatgcgggc agccaggtgg
     7801 cctccaccag cgaggtcttg aaatacacac tgttccaaat cttcagcaag atcgaccgcc
     7861 ctgaagcctc ccgcatcgcc ctgctcctga tggccagcca ggagccccaa cggatgtccc
     7921 ggaactttgt ccgctacgtc cagggcctga agaagaagaa ggtcattgtg atcccggtgg
     7981 gcattgggcc ccatgccaac ctcaagcaga tccgcctcat cgagaagcag gcccctgaga
     8041 acaaggcctt cgtgctgagc agtgtggatg agctggagca gcaaagggac gagatcgtta
     8101 gctacctctg tgaccttgcc cctgaagccc ctcctcctac tctgcccccc cacatggcac
     8161 aagtcactgt gggcccgggg ctcttggggg tttcgaccct ggggcccaag aggaactcca
     8221 tggttctgga tgtggcgttc gtcctggaag gatcggacaa aattggtgaa gccgacttca
     8281 acaggagcaa ggagttcatg gaggaggtga ttcagcggat ggatgtgggc caggacagca
     8341 tccacgtcac ggtgctgcag tactcctaca tggtgaccgt ggagtacccc ttcagcgagg
     8401 cacagtccaa aggggacatc ctgcagcggg tgcgagagat ccgctaccag ggcggcaaca
     8461 ggaccaacac tgggctggcc ctgcggtacc tctctgacca cagcttcttg gtcagccagg
     8521 gtgaccggga gcaggcgccc aacctggtct acatggtcac cggaaatcct gcctctgatg
     8581 agatcaagag gctgcctgga gacatccagg tggtgcccat tggagtgggc cctaatgcca
     8641 acgtgcagga gctggagagg attggctggc ccaatgcccc tatcctcatc caggactttg
     8701 agacgctccc ccgagaggct cctgacctgg tgctgcagag gtgctgctcc ggagaggggc
     8761 tgcagatccc caccctctcc cctgcacctg actgcagcca gcccctggac gtgatccttc
     8821 tcctggatgg ctcctccagt ttcccagctt cttattttga tgaaatgaag agtttcgcca
     8881 aggctttcat ttcaaaagcc aatatagggc ctcgtctcac tcaggtgtca gtgctgcagt
     8941 atggaagcat caccaccatt gacgtgccat ggaacgtggt cccggagaaa gcccatttgc
     9001 tgagccttgt ggacgtcatg cagcgggagg gaggccccag ccaaatcggg gatgccttgg
     9061 gctttgctgt gcgatacttg acttcagaaa tgcatggtgc caggccggga gcctcaaagg
     9121 cggtggtcat cctggtcacg gacgtctctg tggattcagt ggatgcagca gctgatgccg
     9181 ccaggtccaa cagagtgaca gtgttcccta ttggaattgg agatcgctac gatgcagccc
     9241 agctacggat cttggcaggc ccagcaggcg actccaacgt ggtgaagctc cagcgaatcg
     9301 aagacctccc taccatggtc accttgggca attccttcct ccacaaactg tgctctggat
     9361 ttgttaggat ttgcatggat gaggatggga atgagaagag gcccggggac gtctggacct
     9421 tgccagacca gtgccacacc gtgacttgcc agccagatgg ccagaccttg ctgaagagtc
     9481 atcgggtcaa ctgtgaccgg gggctgaggc cttcgtgccc taacagccag tcccctgtta
     9541 aagtggaaga gacctgtggc tgccgctgga cctgcccctg cgtgtgcaca ggcagctcca
     9601 ctcggcacat cgtgaccttt gatgggcaga atttcaagct gactggcagc tgttcttatg
     9661 tcctatttca aaacaaggag caggacctgg aggtgattct ccataatggt gcctgcagcc
     9721 ctggagcaag gcagggctgc atgaaatcca tcgaggtgaa gcacagtgcc ctctccgtcg
     9781 agctgcacag tgacatggag gtgacggtga atgggagact ggtctctgtt ccttacgtgg
     9841 gtgggaacat ggaagtcaac gtttatggtg ccatcatgca tgaggtcaga ttcaatcacc
     9901 ttggtcacat cttcacattc actccacaaa acaatgagtt ccaactgcag ctcagcccca
     9961 agacttttgc ttcaaagacg tatggtctgt gtgggatctg tgatgagaac ggagccaatg
    10021 acttcatgct gagggatggc acagtcacca cagactggaa aacacttgtt caggaatgga
    10081 ctgtgcagcg gccagggcag acgtgccagc ccatcctgga ggagcagtgt cttgtccccg
    10141 acagctccca ctgccaggtc ctcctcttac cactgtttgc tgaatgccac aaggtcctgg
    10201 ctccagccac attctatgcc atctgccagc aggacagttg ccaccaggag caagtgtgtg
    10261 aggtgatcgc ctcttatgcc cacctctgtc ggaccaacgg ggtctgcgtt gactggagga
    10321 cacctgattt ctgtgctatg tcatgcccac catctctggt ctacaaccac tgtgagcatg
    10381 gctgtccccg gcactgtgat ggcaacgtga gctcctgtgg ggaccatccc tccgaaggct
    10441 gtttctgccc tccagataaa gtcatgttgg aaggcagctg tgtccctgaa gaggcctgca
    10501 ctcagtgcat tggtgaggat ggagtccagc accagttcct ggaagcctgg gtcccggacc
    10561 accagccctg tcagatctgc acatgcctca gcgggcggaa ggtcaactgc acaacgcagc
    10621 cctgccccac ggccaaagct cccacgtgtg gcctgtgtga agtagcccgc ctccgccaga
    10681 atgcagacca gtgctgcccc gagtatgagt gtgtgtgtga cccagtgagc tgtgacctgc
    10741 ccccagtgcc tcactgtgaa cgtggcctcc agcccacact gaccaaccct ggcgagtgca
    10801 gacccaactt cacctgcgcc tgcaggaagg aggagtgcaa aagagtgtcc ccaccctcct
    10861 gccccccgca ccgtttgccc acccttcgga agacccagtg ctgtgatgag tatgagtgtg
    10921 cctgcaactg tgtcaactcc acagtgagct gtccccttgg gtacttggcc tcaaccgcca
    10981 ccaatgactg tggctgtacc acaaccacct gccttcccga caaggtgtgt gtccaccgaa
    11041 gcaccatcta ccctgtgggc cagttctggg aggagggctg cgatgtgtgc acctgcaccg
    11101 acatggagga tgccgtgatg ggcctccgcg tggcccagtg ctcccagaag ccctgtgagg
    11161 acagctgtcg gtcgggcttc acttacgttc tgcatgaagg cgagtgctgt ggaaggtgcc
    11221 tgccatctgc ctgtgaggtg gtgactggct caccgcgggg ggactcccag tcttcctgga
    11281 agagtgtcgg ctcccagtgg gcctccccgg agaacccctg cctcatcaat gagtgtgtcc
    11341 gagtgaagga ggaggtcttt atacaacaaa ggaacgtctc ctgcccccag ctggaggtcc
    11401 ctgtctgccc ctcgggcttt cagctgagct gtaagacctc agcgtgctgc ccaagctgtc
    11461 gctgtgagcg catggaggcc tgcatgctca atggcactgt cattgggccc gggaagactg
    11521 tgatgatcga tgtgtgcacg acctgccgct gcatggtgca ggtgggggtc atctctggat
    11581 tcaagctgga gtgcaggaag accacctgca acccctgccc cctgggttac aaggaagaaa
    11641 ataacacagg tgaatgttgt gggagatgtt tgcctacggc ttgcaccatt cagctaagag
    11701 gaggacagat catgacactg aagcgtgatg agacgctcca ggatggctgt gatactcact
    11761 tctgcaaggt caatgagaga ggagagtact tctgggagaa gagggtcaca ggctgcccac
    11821 cctttgatga acacaagtgt ctggctgagg gaggtaaaat tatgaaaatt ccaggcacct
    11881 gctgtgacac atgtgaggag cctgagtgca acgacatcac tgccaggctg cagtatgtca
    11941 aggtgggaag ctgtaagtct gaagtagagg tggatatcca ctactgccag ggcaaatgtg
    12001 ccagcaaagc catgtactcc attgacatca acgatgtgca ggaccagtgc tcctgctgct
    12061 ctccgacacg gacggagccc atgcaggtgg ccctgcactg caccaatggc tctgttgtgt
    12121 accatgaggt tctcaatgcc atggagtgca aatgctcccc caggaagtgc agcaagggac
    12181 cggttagtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg
    12241 gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg
    12301 gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcct tggccaacac
    12361 ttgtcactac tttaacttat ggtgttcaat gcttttcaag atacccagat catatgaaac
    12421 ggcatgactt tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt
    12481 tcaaagatga cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg
    12541 ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttggacaca
    12601 aattggaata caactataac gatcaccagg tgtacatcat ggcagacaaa caaaagaatg
    12661 gaatcaaagc taacttcaaa attagacaca acattgaaga tggaggcgtt caactagcag
    12721 accattatca acaaaatact ccaattggcg atgggcccgt ccttttacca gacaaccatt
    12781 acctgtttac aacttctact ctttcgaaag atcccaacga aaagagagac cacatggtcc
    12841 ttcttgagtt tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaatcaa
    12901 gcggccgcgg aattctgcag tcgacggtac cgcgggcccg ggatccaccg gtcgccacca
    12961 tggcctcctc cgaggacgtc atcaaggagt tcatgcgctt caaggtgcgc atggagggct
    13021 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
    13081 cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc
    13141 tgtcccctca gttccagtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
    13201 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
    13261 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
    13321 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
    13381 tgggctggga ggcctccacc gagcggatgt accccgagga cggcgccctg aagggcgaga
    13441 tcaagatgag gctgaagctg aaggacggcg gccactacga cgccgaggtc aagaccacct
    13501 acatggccaa gaagcccgtg cagctgcccg gcgcctacaa gaccgacatc aagctggaca
    13561 tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgcgcc gagggccgcc
    13621 actccaccgg cgcctaaagc ggccgcgact ctagatcata atcagccata ccacatttgt
    13681 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
    13741 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
    13801 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtg
//
