LOCUS       sequence-339023-       10093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ratiometric biosensor expressed with CMV promoter for probing 
            lysosomal pH in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10093)
  AUTHORS   Chin MY, Patwardhan AR, Ang KH, Wang AL, Alquezar C, Welch M, Nguyen
            PT, Grabe M, Molofsky AV, Arkin MR, Kao AW
  TITLE     Genetically Encoded, pH-Sensitive mTFP1 Biosensor for Probing 
            Lysosomal pH.
  JOURNAL   ACS Sens. 2021 Jun 8. doi: 10.1021/acssensors.0c02318.
  PUBMED    34102054
REFERENCE   2  (bases 1 to 10093)
  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: "ACS Sens. 
            2021 Jun 8. doi: 10.1021/acssensors.0c02318."
COMMENT     Sequence Label: pLJM1-FIRE-pHLy
FEATURES             Location/Qualifiers
     source          1..10093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        131..334
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     285..305
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      373..382
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      377..386
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             463..1167
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="VSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDGT
                     NTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTFE
                     DKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLKG
                     DVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESAV
                     ARNSTDGMDELYK"
     CDS             2398..3105
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2540..2558)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    3176..3293
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3342..3852
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3771..3789
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3874..4473
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3874..3893)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4370..4390
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4601..4834
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4906..5027
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4943..4962)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4997..5016
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5067..5202
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5222..5241)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5223..5241)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5251..5268)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5251..5267)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5260..5282)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5409..5864
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5496..5515)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5706..5727
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5890..5994
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5995..6063,6064..6855)
                     /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     complement(6213..6232)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7026..7614
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7515..7534
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7768..7785
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7902..7923
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7938..7943,7944..7961,7962..7968)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7976..7992
                     /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)."
     primer_bind     7981..8003
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8000..8016
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8000..8016
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8035..8055
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8037..8055
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8083..8309
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8310..8490
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8537..8662
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9155..9388
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9573..9617
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     primer_bind     9915..9935
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
       61 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      121 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      181 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      241 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      301 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      361 gcgctaccgg tcgccaccat ggcggccccc ggcagcgccc ggcgacccct gctgctgcta
      421 ctgctgttgc tgctgctcgg cctcatgcat tgtgcgtcag cagtgagcaa gggcgaggag
      481 accacaatgg gcgtaatcaa gcccgacatg aagatcaagc tgaagatgga gggcaacgtg
      541 aatggccacg ccttcgtgat cgagggcgag ggcgagggca agccctacga cggcaccaac
      601 accatcaacc tggaggtgaa ggagggagcc cccctgccct tctcctacga cattctgacc
      661 accgcgttcg cctacggcaa cagggccttc accaagtacc ccgacgacat ccccaactac
      721 ttcaagcagt ccttccccga gggctactct tgggagcgca ccatgacctt cgaggacaag
      781 ggcatcgtga aggtgaagtc cgacatctcc atggaggagg actccttcat ctacgagata
      841 cacctcaagg gcgagaactt cccccccaac ggccccgtga tgcagaagaa gaccaccggc
      901 tgggacgcct ccaccgagag gatgtacgtg cgcgacggcg tgctgaaggg cgacgtcaag
      961 cacaagctgc tgctggaggg cggcggccac caccgcgttg acttcaagac catctacagg
     1021 gccaagaagg cggtgaagct gcccgactat cactttgtgg accaccgcat cgagatcctg
     1081 aaccacgaca aggactacaa caaggtgacc gtttacgaga gcgccgtggc ccgcaactcc
     1141 accgacggca tggacgagct gtacaagggc ggcagcggcg gcggcagcgg cagcggcggc
     1201 ggcagcggcg caatgtttat ggtgaaaaat ggcaacggga ccgcgtgcat aatggccaac
     1261 ttctctgctg ccttctcagt gaactacgac accaagagtg gccctaagaa catgaccttt
     1321 gacctgccat cagatgccac agtggtgctc aaccgcagct cctgtggaaa agagaacact
     1381 tctgacccca gtctcgtgat tgcttttgga agaggacata cactcactct caatttcacg
     1441 agaaatgcaa cacgttacag cgtccagctc atgagttttg tttataactt gtcagacaca
     1501 caccttttcc ccaatgcgag ctccaaagaa atcaagactg tggaatctat aactgacatc
     1561 agggcagata tagataaaaa atacagatgt gttagtggca cccaggtcca catgaacaac
     1621 gtgaccgtaa cgctccatga tgccaccatc caggcgtacc tttccaacag cagcttcagc
     1681 aggggagaga cacgctgtga acaagacagg ccttccccaa ccacagcgcc ccctgcgcca
     1741 cccagcccct cgccctcacc cgtgcccaag agcccctctg tggacaagta caacgtgagc
     1801 ggcaccaacg ggacctgcct gctggccagc atggggctgc agctgaacct cacctatgag
     1861 aggaaggaca acacgacggt gacaaggctt ctcaacatca accccaacaa gacctcggcc
     1921 agcgggagct gcggcgccca cctggtgact ctggagctgc acagcgaggg caccaccgtc
     1981 ctgctcttcc agttcgggat gaatgcaagt tctagccggt ttttcctaca aggaatccag
     2041 ttgaatacaa ttcttcctga cgccagagac cccgccttta aagctgccaa cggctccctg
     2101 cgagcgctgc aggccacagt cggcaattcc tacaagtgca acgcggagga gcacgtccgt
     2161 gtcacgaagg cgttttcagt caatatattc aaagtgtggg tccaggcttt caaggtggaa
     2221 ggtggccagt ttggctctgt ggaggagtgt ctgctggacg agaacagcat gctgatcccc
     2281 atcgctgtgg gtggtgccct ggcggggctg gtcctcatcg tcctcatcgc ctacctcgtc
     2341 ggcaggaaga ggagtcacgc aggctaccag actatcccgg cgccggcgcc ggcgccggtg
     2401 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     2461 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     2521 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     2581 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     2641 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     2701 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     2761 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     2821 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     2881 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     2941 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     3001 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     3061 gagggccgcc actccaccgg cggcatggac gagctgtaca agtagtccgg actcagatct
     3121 cgagctcaag cttcgaattc tcgacctcga gacaaatggc agtattcatc cacaatttta
     3181 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa
     3241 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt
     3301 attacaggga cagcagagat ccactttggc cgcggctcga gggggttggg gttgcgcctt
     3361 ttccaaggca gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa
     3421 cgcagcggcg ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg
     3481 atcttcgccg ctacccttgt gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg
     3541 gaaggttcct tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact
     3601 agtaccctcg cagacggaca gcgccaggga gcaatggcag cgcgccgacc gcgatgggct
     3661 gtggccaata gcggctgctc agcagggcgc gccgagagca gcggccggga aggggcggtg
     3721 cgggaggcgg ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca
     3781 ttctgcaagc ctccggagcg cacgtcggca gtcggctccc tcgttgaccg aatcaccgac
     3841 ctctctcccc agggggatcc accggagctt accatgaccg agtacaagcc cacggtgcgc
     3901 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     3961 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     4021 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     4081 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     4141 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     4201 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     4261 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     4321 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     4381 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     4441 acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg
     4501 accgaaagga gcgcacgacc ccatgcatcg gtacctttaa gaccaatgac ttacaaggca
     4561 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
     4621 caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga
     4681 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     4741 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     4801 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
     4861 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     4921 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4981 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     5041 tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5101 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     5161 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc
     5221 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     5281 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     5341 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     5401 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     5461 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     5521 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     5581 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     5641 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5701 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     5761 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     5821 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     5881 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5941 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6001 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     6061 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     6121 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6181 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     6241 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6301 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6361 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6421 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6481 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     6541 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6601 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6661 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6721 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6781 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6841 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6901 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6961 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7021 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     7081 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     7141 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     7201 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7261 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7321 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7381 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7441 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7501 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     7561 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7621 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     7681 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     7741 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     7801 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     7861 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     7921 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     7981 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     8041 aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac
     8101 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
     8161 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
     8221 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
     8281 tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct
     8341 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     8401 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     8461 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
     8521 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     8581 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     8641 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     8701 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     8761 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     8821 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     8881 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     8941 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     9001 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     9061 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     9121 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     9181 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     9241 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     9301 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     9361 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     9421 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     9481 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     9541 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     9601 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     9661 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     9721 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     9781 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     9841 gatccattcg attagtgaac ggatctcgac ggtatcgatc acgagactag cctcgagcgg
     9901 ccgccccctt caccgagggc ctatttccca tgattccttc atatttgcat atacgataca
     9961 aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa
    10021 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
    10081 aaatggacta tca
//
