LOCUS       sequence-90400-e       10058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the pH sensor ratiometric pHluorin in the mitochondria of 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10058)
  AUTHORS   Venkatachalam V, Cohen AE
  TITLE     Imaging GFP-Based Reporters in Neurons with Multiwavelength 
            Optogenetic Control.
  JOURNAL   Biophys J. 2014 Oct 7;107(7):1554-63. doi: 
            10.1016/j.bpj.2014.08.020.
  PUBMED    25296307
REFERENCE   2  (bases 1 to 10058)
  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"; doi: 
            "10.1016/j.bpj.2014.08"; journalName: "Biophys J"; date: "2014-10-7-
            7"; volume: "107"; issue: "7"; pages: "1554-63"
COMMENT     Sequence Label: VV064: 1xCox8 - ratiometric pHluorin in fck
FEATURES             Location/Qualifiers
     source          1..10058
                     /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        2602..3890
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     CDS             3906..3980
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             4014..4727
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Miesenbock et al., 1998)"
                     /label=ratiometric pHluorin
                     /note="responds to a drop in pH with a reversible decrease 
                     in excitation at 395 nm and a corresponding increase in 
                     excitation at 475 nm"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKDDGNILGHKLEYNYNEHLVYIMADKQKNGTKAI
                     FQVHHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLHTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(4041..4069)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4664..4684
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    4750..5338
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4803..4823)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5221..5232)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5341..5357)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5342..5358)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5863..6043
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6069..6086)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6075..6299
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6345..6773
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6432..6451)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6642..6663
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6782..6802)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6787..7116
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6967..7102
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7029..7048
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7164..7211
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7230..7604
                     /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    7734..7855
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7771..7790)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7825..7844
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7904..7920)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7904..7920)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7917..7939)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7928..7944
                     /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(7952..7958,7959..7976,7977..7982))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7997..8018
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8135..8152)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8306..8894)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8386..8405)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9065..9856,9857..9925))
                     /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     9688..9707
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9926..10030)
                     /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 acattatggc cttaggtcac ttcatctcca tggggttctt
     2641 cttctgattt tctagaaaat gagatggggg tgcagagagc ttcctcagtg acctgcccag
     2701 ggtcacatca gaaatgtcag agctagaact tgaactcaga ttactaatct taaattccat
     2761 gccttggggg catgcaagta cgatatacag aaggagtgaa ctcattaggg cagatgacca
     2821 atgagtttag gaaagaagag tccagggcag ggtacatcta caccacccgc ccagccctgg
     2881 gtgagtccag ccacgttcac ctcattatag ttgcctctct ccagtcctac cttgacggga
     2941 agcacaagca gaaactggga caggagcccc aggagaccaa atcttcatgg tccctctggg
     3001 aggatgggtg gggagagctg tggcagaggc ctcaggaggg gccctgctgc tcagtggtga
     3061 cagatagggg tgagaaagca gacagagtca ttccgtcagc attctgggtc tgtttggtac
     3121 ttcttctcac gctaaggtgg cggtgtgata tgcacaatgg ctaaaaagca gggagagctg
     3181 gaaagaaaca aggacagaga cagaggccaa gtcaaccaga ccaattccca gaggaagcaa
     3241 agaaaccatt acagagacta caagggggaa gggaaggaga gatgaattag cttcccctgt
     3301 aaaccttaga acccagctgt tgccagggca acggggcaat acctgtctct tcagaggaga
     3361 tgaagttgcc agggtaacta catcctgtct ttctcaagga ccatcccaga atgtggcacc
     3421 cactagccgt taccatagca actgcctctt tgccccactt aatcccatcc cgtctgttaa
     3481 aagggcccta tagttggagg tgggggaggt aggaagagcg atgatcactt gtggactaag
     3541 tttgttcaca tccccttctc caaccccctc agtacatcac cctgggggaa cagggtccac
     3601 ttgctcctgg gcccacacag tcctgcagta ttgtgtatat aaggccaggg caaagaggag
     3661 caggttttaa agtgaaaggc aggcaggtgt tggggaggca gttaccgggg caacgggaac
     3721 agggcgtttc ggaggtggtt gccatgggga cctggatgct gacgaaggct cgcgaggctg
     3781 tgagcagcca cagtgccctg ctcagaagcc ccaagctcgt cagtcaagcc ggttctccgt
     3841 ttgcactcag gagcacgggc aggcgagtgg cccctagttc tgggggcagc gggggatccg
     3901 ccaccatgtc cgtcctgacg ccgctgctgc tgcggggctt gacaggctcg gcccggcggc
     3961 tcccagtgcc gcgcgccaag atccattcgt tgggtgaccc gccagtcgct agcagtaaag
     4021 gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg
     4081 ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga aaacttaccc
     4141 ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt gtcactactt
     4201 tctcttatgg tgttcaatgc ttttcaagat acccagatca tatgaaacgg catgactttt
     4261 tcaagagtgc catgcccgaa ggttatgtac aggaaagaac tatatttttc aaagatgacg
     4321 ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg
     4381 agttaaaagg tattgatttt aaagatgatg gaaacattct tggacacaaa ttggaataca
     4441 actataacga gcacttggtg tacatcatgg cagacaaaca aaagaatggt accaaagcta
     4501 tctttcaagt tcaccacaac attgaagatg gaggcgttca actagcagac cattatcaac
     4561 aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac ctgcacacac
     4621 aatctgccct ttcgaaagat cccaacgaaa agagagacca catggtcctt cttgagtttg
     4681 taacagctgc tgggattaca catggcatgg atgaactata caaatgaatt cgatatcaag
     4741 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     4801 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     4861 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     4921 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     4981 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     5041 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     5101 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     5161 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     5221 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     5281 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     5341 cgataccgtc gacctcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca
     5401 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca
     5461 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     5521 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc
     5581 cttgatctgt ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca
     5641 gggccaggga tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag
     5701 caagagaagg tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc
     5761 ctgcatggga tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta
     5821 gcatttcatc acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac
     5881 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     5941 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     6001 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagggcccg tttaaacccg
     6061 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     6121 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     6181 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     6241 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     6301 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     6361 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     6421 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     6481 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     6541 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     6601 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     6661 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     6721 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     6781 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     6841 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     6901 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     6961 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     7021 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     7081 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     7141 attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag
     7201 tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc
     7261 tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc
     7321 gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca
     7381 gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc
     7441 tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg
     7501 ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc
     7561 cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt
     7621 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     7681 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     7741 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     7801 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     7861 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     7921 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     7981 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     8041 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     8101 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     8161 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     8221 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     8281 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     8341 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     8401 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     8461 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     8521 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     8581 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     8641 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     8701 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     8761 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     8821 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     8881 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     8941 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     9001 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     9061 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     9121 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     9181 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     9241 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     9301 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     9361 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     9421 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     9481 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     9541 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     9601 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     9661 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     9721 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     9781 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     9841 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     9901 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     9961 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
    10021 gggttccgcg cacatttccc cgaaaagtgc cacctgac
//
