LOCUS       sequence-444921-        8484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  R2pH-LAMP1-labelled lysosomes with FRB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8484)
  AUTHORS   Cheetham-Wilkinson IJ, Sivalingam B, Flitton C, Flottmann F, Vehling
            L, Drechsler M, Stojchevska M, Raimondi A, Paululat A, Frohlich F, 
            Swan LE, Stagi M
  TITLE     RpH-ILV: Probe for lysosomal pH and acute LLOMe-induced membrane 
            permeabilization in cell lines and Drosophila.
  JOURNAL   Sci Adv. 2025 Jan 3;11(1):eadr7325. doi: 10.1126/sciadv.adr7325. 
            Epub 2025 Jan 3.
  PUBMED    39752501
REFERENCE   2  (bases 1 to 8484)
  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: "Sci Adv."; 
            date: "2025-01-3"; volume: "11(1):eadr7325. doi"; pages: " 
            10.1126/sciadv.adr7325. Epub 2025 Jan 3"
COMMENT     Sequence Label: R2pH-LAMP-FRB
FEATURES             Location/Qualifiers
     source          1..8484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    828..962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(852..871)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     906..925
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(968..1163)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      982..1117
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    1222..1243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1258..1263,1264..1281,1282..1288)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1296..1312
                     /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     1301..1323
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1320..1336
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1320..1336
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1679..1698
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1697..1752
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1798..1817
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(1904..2179)
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="LWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLK
                     ETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISK"
     CDS             complement(3338..4051)
                     /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="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3378..3398)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     3993..4021
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     CDS             complement(4073..4780)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             complement(4793..5500)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     regulatory      5574..5583
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5612..5631)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          5639..6655
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(5693..5715)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     enhancer        6939..7318
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7349..7453
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7454..7522,7523..8314)
                     /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(7672..7691)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagcggat ccagacatga
      841 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      901 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      961 ttagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     1021 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     1081 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     1141 tggttgctga ctaattgaga tgcggatccg ctggcacgac aggtttcccg actggaaagc
     1201 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     1261 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     1321 aggaaacagc tatgaccatg attacgccaa gcttgggctg caggtcgagg gatctccata
     1381 agagaagagg gacagctatg actgggagta gtcaggagag gaggaaaaat ctggctagta
     1441 aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa taacacaaaa caaaatataa
     1501 aaaaaatcta acctcaagtc aaggcttttc tatggaataa ggaatggaca gcagggggct
     1561 gtttcatata ctgatgacct ctttatagcc aacctttgtt catggcagcc agcatatggg
     1621 catatgttgc caaactctaa accaaatact cattctgatg ttttaaatga tttgccctcc
     1681 catatgtcct tccgagtgag agacacaaaa aattccaaca cactattgca atgaaaataa
     1741 atttccttta ttagccagaa gtcagatgct caaggggctt catgatgtcc ccataatttt
     1801 tggcagaggg aaaaagatct cagtggtatt tgtgagccag ggcattggcc acaccagcca
     1861 ccaccttctg ataggcagcc tgcacctgag gagtgaattc ctactttgag attcgtcgga
     1921 acacatgata atagaggtcc caggcttggg tgaggtcctt gacattccct gatttcatgt
     1981 acttcctgca ccactcttgg gcctccatta aatctcgacc ataggcctga ttaaaggatg
     2041 tttccttcag agtctggggg ccccgttcca tcatagcatg caagggctcc agcacctcaa
     2101 acatgccttt cacgttcctt tccccaaagt acaaacgaga tgcttcttcc aggccttcat
     2161 gccacatctc atgccagagg acgatggtct gatagccggc gtgactcctc ttcctgccaa
     2221 tgaggtaggc aatgaggacg atgaggacca gccctgccag ggcaccgccc acagcaatgg
     2281 ggatcaacat gttgttacca tcctgaacac actcttccac agacccaaac ctgtcactgt
     2341 ccaccttgaa agcctggacc tgcacactga agacattgag ggagagcatc ttgctgacaa
     2401 agatgtgttc ctcagtgttg cacttgtatg agtttcccac agtggcctga agagctttca
     2461 gtgaatggtt ggagatgctg aatgtgggca ctagggcatc aggaagagtc atattcaagc
     2521 gcactccttg caggaaaaac aggctagagc tggcattcat cccaaactgc aattccaggg
     2581 ctctgttctt gttctccact ttcagggtca ccaagttgat accgcaactc ccactagatg
     2641 tgtcatttgg gctgatgttg aacgctctgg tcaccgtctt gttgtccttt ttcaggtagg
     2701 tgatattcag ttgcagtgcc atagaggcca gcaggcaggt tccgttgtta ccagtaacat
     2761 tgtacttgga tacagtgggg tttgtgggca caagtggtgg tgaggggctg ggtggcccag
     2821 tggttgggga aggtccatcc tgtgtgcagt gtgtctcttc cttgctgaag ttgccactcg
     2881 acaggtaggc ctggatagtg gcatcccgga gcacaacggt cacattcttc atgtagaccc
     2941 ggatatcact gacacaccgg tatgctttgt tgatgtctgc cttgatgtca gttgtggaat
     3001 ccatggtgta gatctctttg ctgatggcat tgggaaaatg ttctgtatct gacaagttat
     3061 atgtaaaata catatgctgg acactgtaac gtgttgtatt ttttgtgaag ttgagtgtca
     3121 gtaaatatcc tcttccaaaa gtaattgtga ggctggggtc agaaacattt tctttaccac
     3181 aagaactgcc atttttcagt acttctgcag aggctggcag ggaaatgttc acgatctgag
     3241 aaccattcgc agtctcgtag gtggtcagaa aggaggcaga gaagctggcc attatacacg
     3301 ttgtgccatt gtttttcacc tcaaagagcc ccccgggttt gtatagttca tccatgccat
     3361 gtgtaatccc agcagctgtt acaaactcaa gaaggaccat gtggtctctc ttttcgttgg
     3421 gatctttcga aagagtagaa gttgtaaaca ggtaatggtt gtctggtaaa aggacgggcc
     3481 catcgccaat tggagtattt tgttgataat ggtctgctag ttgaacgcct ccatcttcaa
     3541 tgttgtgtct aattttgaag ttagctttga ttccattctt ttgtttgtct gccatgatgt
     3601 acacctggtg atcgttatag ttgtattcca atttgtgtcc aagaatgttt ccatcttctt
     3661 taaaatcaat accttttaac tcgattctat taacaagggt atcaccttca aacttgactt
     3721 cagcacgtgt cttgtagttc ccgtcatctt tgaaaaatat agttctttcc tgtacataac
     3781 cttcgggcat ggcactcttg aaaaagtcat gccgtttcat atgatctggg tatcttgaaa
     3841 agcattgaac gccataagtt aaagtagtga caagtgttgg ccaaggaaca ggtagttttc
     3901 cagtagtgca aataaattta agggtaagtt ttccgtatgt tgcatcacct tcaccctctc
     3961 cactgacaga aaatttgtgc ccattaacat caccatctaa ttcaacaaga attgggacaa
     4021 ctccagtgaa aagttcttct cctttactca tcccgggccc cccgctagct cccttgtaca
     4081 gctcgtccat gccgccggtg gagtggcggc cctcggcgcg ttcgtactgt tccacgatgg
     4141 tgtagtcctc gttgtgggag gtgatgtcca acttgatgtt gacgttgtag gcgccgggca
     4201 gctgcacggg cttcttggcc ttgtaggtgg tcttgacctc agcgtcgtag tggccgccgt
     4261 ccttcagctt cagcctctgc ttgatctcgc ccttcagggc gccgtcctcg gggtacatcc
     4321 gctcggagga ggcctcccag cccatggtct tcttctgcat tacggggccg tcggagggga
     4381 agttggtgcc gcgcagcttc accttgtaga tgaactcgcc gtcctgcagg gaggagtcct
     4441 gggtcacggt caccacgccg ccgtcctcga agttcatcac gcgctcccac ttgaagccct
     4501 cggggaagga cagcttcaag tagtcgggga tgtcggcggg gtgcttcacg taggccttgg
     4561 agccgtacat gaactgaggg gacaggatgt cccaggcgaa gggcaggggg ccacccttgg
     4621 tcaccttcag cttggcggtc tgggtgccct cgtaggggcg gccctcgccc tcgccctcga
     4681 tctcgaactc gtggccgttc acggagccct ccatgtgcac cttgaagcgc atgaactcct
     4741 tgatgatggc catgttatcc tcctcgccct tgctcaccat accgctagct cccttgtaca
     4801 gctcgtccat gccgccggtg gagtggcggc cctcggcgcg ttcgtactgt tccacgatgg
     4861 tgtagtcctc gttgtgggag gtgatgtcca acttgatgtt gacgttgtag gcgccgggca
     4921 gctgcacggg cttcttggcc ttgtaggtgg tcttgacctc agcgtcgtag tggccgccgt
     4981 ccttcagctt cagcctctgc ttgatctcgc ccttcagggc gccgtcctcg gggtacatcc
     5041 gctcggagga ggcctcccag cccatggtct tcttctgcat tacggggccg tcggagggga
     5101 agttggtgcc gcgcagcttc accttgtaga tgaactcgcc gtcctgcagg gaggagtcct
     5161 gggtcacggt caccacgccg ccgtcctcga agttcatcac gcgctcccac ttgaagccct
     5221 cggggaagga cagcttcaag tagtcgggga tgtcggcggg gtgcttcacg taggccttgg
     5281 agccgtacat gaactgaggg gacaggatgt cccaggcgaa gggcaggggg ccacccttgg
     5341 tcaccttcag cttggcggtc tgggtgccct cgtaggggcg gccctcgccc tcgccctcga
     5401 tctcgaactc gtggccgttc acggagccct ccatgtgcac cttgaagcgc atgaactcct
     5461 tgatgatggc catgttatcc tcctcgccct tgctcaccat accgctagct gctgaggcgc
     5521 catgtgcaag gcctgccagc agcagcagga gcagcggccg ccgggcgccg ggggccgcca
     5581 tggtgaattc tttgccaaaa tgatgagaca gcacaataac cagcacgttg cccaggagct
     5641 gtaggaaaaa gaagaaggca tgaacatggt tagcagaggc tctagagccg ccggtcacac
     5701 gccagaagcc gaaccccgcc ctgccccgtc ccccccgaag gcagccgtcc ccccgcggac
     5761 agccccgagg ctggagaggg agaaggggac ggcggcgcgg cgacgcacga aggccctccc
     5821 cgcccatttc cttcctgccg gcgccgcacc gcttcgcccc gcgcccgcta gagggggtgc
     5881 ggcggcgcct cccagatttc ggctccgcac agatttggga caaaggaagt ccctgcgccc
     5941 tctcgcacga ttaccataaa aggcaatggc tgcggctcgc cgcgcctcga cagccgccgg
     6001 cgctccgggg gccgccgcgc cccycccccg agcccycccc ggcccgrggc ggccccgccc
     6061 cgcccggcac ccccmcctgc cgccaccccc cgcccggcac ggcgagcccc gcgccacgcc
     6121 ccgtacggag ccccgcaccc gaagccgggc cgtgctcagc aactcgggga ggggggtgca
     6181 gggggggttg cagcccgacc gacgcgccca caccccctgc tcaccccccc acgcacacac
     6241 cccgcacgca gcctttgttc ccctcgcagc cccccccgca ccgcggggca ccgcccccgg
     6301 ccgcgctccc ctcgcgcaca ctgcggagcg cacaaagccc cgcgccgcgc ccgcagcgct
     6361 cacagccgcc gggcagcgcg gagccgcacg cggcgctccc cacgcacaca cacacgcacg
     6421 caccccccga gccgctcccc ccgcacaaag ggccctcccg gagcccctca aggctttcac
     6481 gcagccacag aaaagaaaca agccgtcatt aaaccaagcg ctaattacag cccggaggag
     6541 aagggccgtc ccgcccgctc acctgtggga gtaacgcggt cagtcagagc cggggcgggc
     6601 ggcgcgaggc ggcggcggag cggggcacgg ggcgaaggca gcgcgcagcg actcccgccc
     6661 gccgcgcgct tcgcttttta tagggccgcc gccgccgccg cctcgccata aaaggaaact
     6721 ttcggagcgc gccgctctga ttggctgccg ccgcmccycy ccgccycscc ccsccccscc
     6781 cccccccccc cccccccccc cccsgcgcgc gccccccccc cccccccccc cgcccccatc
     6841 gctgcacaaa ataattaaaa aataaataaa tacaaaattg ggggtgggga ggggggggag
     6901 atggggagag tgaagcagaa cgtggggctc acctcgacca tggtaatagc gatgactaat
     6961 acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag
     7021 gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga
     7081 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
     7141 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
     7201 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
     7261 ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcga
     7321 cccaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     7381 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     7441 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     7501 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     7561 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     7621 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     7681 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     7741 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     7801 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     7861 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     7921 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     7981 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     8041 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     8101 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     8161 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     8221 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     8281 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     8341 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     8401 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     8461 cgtagaaaag atcaaaggat cttc
//
