LOCUS       sequence-331211-       11962 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ratiometric biosensor expressed with UbC promoter for probing 
            lysosomal pH in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11962)
  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 11962)
  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: pFUGW-FIRE-pHLy
FEATURES             Location/Qualifiers
     source          1..11962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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"
     misc_feature    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2622..3833
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3670..3689
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      3881..3890
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3885..3894
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3971..4675
                     /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             5906..6613
                     /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(6048..6066)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    6654..7242
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6707..6727)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7125..7136)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7245..7261)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7246..7262)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7767..7947
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7973..7990)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7979..8203
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8249..8677
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8336..8355)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8546..8567
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8686..8706)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8691..9020
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8871..9006
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8933..8952
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9068..9115
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9134..9508
                     /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    9638..9759
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9675..9694)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9729..9748
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9808..9824)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9808..9824)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9821..9843)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9832..9848
                     /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(join(9856..9862,9863..9880,9881..9886))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9901..9922
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10039..10056)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10210..10798)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10290..10309)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10969..11760,11761..11829))
                     /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     11592..11611
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11830..11934)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta acccgtgtcg gctccagatc tggcctccgc gccgggtttt
     2641 ggcgcctccc gcgggcgccc ccctcctcac ggcgagcgct gccacgtcag acgaagggcg
     2701 cagcgagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga
     2761 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc
     2821 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg
     2881 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc
     2941 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga
     3001 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc
     3061 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt
     3121 ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt
     3181 cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg
     3241 ggggcatggt gggcggcaag aacccaaggt cttgaggcct tcgctaatgc gggaaagctc
     3301 ttattcgggt gagatgggct ggggcaccat ctggggaccc tgacgtgaag tttgtcactg
     3361 actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt gccgttgggc
     3421 agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt
     3481 tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct tttctccgtc
     3541 gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg ccggacctct
     3601 ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt acctatcttc
     3661 ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg
     3721 aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat tttcagtgtt
     3781 agactagtaa attgtccgct aaattctggc cgtttttggc ttttttgtta gacgaagctt
     3841 gggctgcagg tcgactctag aggatccccg ggtaccggtc gccaccatgg cggcccccgg
     3901 cagcgcccgg cgacccctgc tgctgctact gctgttgctg ctgctcggcc tcatgcattg
     3961 tgcgtcagca gtgagcaagg gcgaggagac cacaatgggc gtaatcaagc ccgacatgaa
     4021 gatcaagctg aagatggagg gcaacgtgaa tggccacgcc ttcgtgatcg agggcgaggg
     4081 cgagggcaag ccctacgacg gcaccaacac catcaacctg gaggtgaagg agggagcccc
     4141 cctgcccttc tcctacgaca ttctgaccac cgcgttcgcc tacggcaaca gggccttcac
     4201 caagtacccc gacgacatcc ccaactactt caagcagtcc ttccccgagg gctactcttg
     4261 ggagcgcacc atgaccttcg aggacaaggg catcgtgaag gtgaagtccg acatctccat
     4321 ggaggaggac tccttcatct acgagataca cctcaagggc gagaacttcc cccccaacgg
     4381 ccccgtgatg cagaagaaga ccaccggctg ggacgcctcc accgagagga tgtacgtgcg
     4441 cgacggcgtg ctgaagggcg acgtcaagca caagctgctg ctggagggcg gcggccacca
     4501 ccgcgttgac ttcaagacca tctacagggc caagaaggcg gtgaagctgc ccgactatca
     4561 ctttgtggac caccgcatcg agatcctgaa ccacgacaag gactacaaca aggtgaccgt
     4621 ttacgagagc gccgtggccc gcaactccac cgacggcatg gacgagctgt acaagggcgg
     4681 cagcggcggc ggcagcggca gcggcggcgg cagcggcgca atgtttatgg tgaaaaatgg
     4741 caacgggacc gcgtgcataa tggccaactt ctctgctgcc ttctcagtga actacgacac
     4801 caagagtggc cctaagaaca tgacctttga cctgccatca gatgccacag tggtgctcaa
     4861 ccgcagctcc tgtggaaaag agaacacttc tgaccccagt ctcgtgattg cttttggaag
     4921 aggacataca ctcactctca atttcacgag aaatgcaaca cgttacagcg tccagctcat
     4981 gagttttgtt tataacttgt cagacacaca ccttttcccc aatgcgagct ccaaagaaat
     5041 caagactgtg gaatctataa ctgacatcag ggcagatata gataaaaaat acagatgtgt
     5101 tagtggcacc caggtccaca tgaacaacgt gaccgtaacg ctccatgatg ccaccatcca
     5161 ggcgtacctt tccaacagca gcttcagcag gggagagaca cgctgtgaac aagacaggcc
     5221 ttccccaacc acagcgcccc ctgcgccacc cagcccctcg ccctcacccg tgcccaagag
     5281 cccctctgtg gacaagtaca acgtgagcgg caccaacggg acctgcctgc tggccagcat
     5341 ggggctgcag ctgaacctca cctatgagag gaaggacaac acgacggtga caaggcttct
     5401 caacatcaac cccaacaaga cctcggccag cgggagctgc ggcgcccacc tggtgactct
     5461 ggagctgcac agcgagggca ccaccgtcct gctcttccag ttcgggatga atgcaagttc
     5521 tagccggttt ttcctacaag gaatccagtt gaatacaatt cttcctgacg ccagagaccc
     5581 cgcctttaaa gctgccaacg gctccctgcg agcgctgcag gccacagtcg gcaattccta
     5641 caagtgcaac gcggaggagc acgtccgtgt cacgaaggcg ttttcagtca atatattcaa
     5701 agtgtgggtc caggctttca aggtggaagg tggccagttt ggctctgtgg aggagtgtct
     5761 gctggacgag aacagcatgc tgatccccat cgctgtgggt ggtgccctgg cggggctggt
     5821 cctcatcgtc ctcatcgcct acctcgtcgg caggaagagg agtcacgcag gctaccagac
     5881 tatcccggcg ccggcgccgg cgccggtgag caagggcgag gaggataaca tggccatcat
     5941 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
     6001 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     6061 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     6121 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     6181 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     6241 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     6301 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
     6361 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
     6421 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     6481 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
     6541 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     6601 gctgtacaag tagagcggcc gcgactctag aattcgatat caagcttatc gataatcaac
     6661 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     6721 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     6781 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     6841 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     6901 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     6961 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     7021 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     7081 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     7141 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     7201 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc
     7261 gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt
     7321 gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac
     7381 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     7441 ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct
     7501 accacacaca aggctacttc cctgattggc agaactacac accagggcca gggatcagat
     7561 atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag aaggtagaag
     7621 aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat gggatggatg
     7681 acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt catcacatgg
     7741 cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc tgagcctggg
     7801 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     7861 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     7921 tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc agcctcgact
     7981 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     8041 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     8101 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     8161 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     8221 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     8281 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     8341 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     8401 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     8461 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     8521 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     8581 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     8641 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     8701 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     8761 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     8821 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     8881 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     8941 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     9001 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     9061 agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     9121 gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt
     9181 cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga
     9241 cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca
     9301 ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc
     9361 cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga
     9421 gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt
     9481 gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc
     9541 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     9601 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     9661 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     9721 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     9781 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     9841 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     9901 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     9961 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
    10021 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
    10081 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
    10141 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
    10201 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
    10261 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
    10321 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
    10381 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
    10441 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
    10501 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
    10561 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
    10621 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
    10681 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
    10741 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
    10801 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
    10861 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
    10921 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
    10981 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
    11041 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
    11101 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
    11161 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
    11221 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
    11281 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
    11341 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
    11401 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
    11461 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
    11521 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
    11581 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
    11641 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
    11701 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
    11761 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
    11821 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
    11881 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
    11941 tccccgaaaa gtgccacctg ac
//
