LOCUS       sequence-274013-        9169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  final optimized bioluminescent reporter for detecting extracellular 
            pH changes in tumor acidosis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9169)
  AUTHORS   Ong TT, Ang Z, Verma R, Koean R, Tam JKC, Ding JL
  TITLE     pHLuc, a Ratiometric Luminescent Reporter for in vivo Monitoring of 
            Tumor Acidosis.
  JOURNAL   Front Bioeng Biotechnol. 2020 May 8;8:412. doi: 
            10.3389/fbioe.2020.00412. eCollection 2020.
  PUBMED    32457886
REFERENCE   2  (bases 1 to 9169)
  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: "Front 
            Bioeng Biotechnol."; date: "2020-05-8"; volume: "8:412. doi"; pages:
            " 10.3389/fbioe.2020.00412. eCollection 2020"
COMMENT     Sequence Label: pHLuc (Antares-SEPLuc C1A4E+6)
FEATURES             Location/Qualifiers
     source          1..9169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..55)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    44..268
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      314..742
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(401..420)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     611..632
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(751..771)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        756..1085
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      936..1071
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     998..1017
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    1325..1446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1362..1381)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1416..1435
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1495..1511)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1495..1511)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1508..1530)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1519..1535
                     /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(1543..1549,1550..1567,1568..1573))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1588..1609
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1726..1743)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1897..2482)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1977..1996)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2653..3444,3445..3513))
                     /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     3276..3295
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3514..3618)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3693..3712)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3884..4263
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4264..4467
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4418..4438
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4512..4531
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4512..4530
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5291..5797
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="FTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRIV
                     LSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVTP
                     NMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWRL
                     CERILA"
     CDS             6497..6550
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             6557..7156
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHLER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6557..6576)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7053..7073
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7158..7731
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7325..7342)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7552..7571
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
ORIGIN
        1 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
       61 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
      121 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
      181 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
      241 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
      301 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
      361 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
      421 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
      481 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
      541 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
      601 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
      661 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
      721 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
      781 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
      841 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
      901 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
      961 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     1021 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     1081 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     1141 gatcgtttcg cgcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     1201 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     1261 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     1321 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     1381 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     1441 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     1501 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     1561 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1621 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1681 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1741 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1801 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1861 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1921 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1981 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2041 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2101 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2161 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2221 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2281 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2341 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2401 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     2461 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2521 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2581 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2641 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2701 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     2761 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     2821 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     2881 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     2941 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3001 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3061 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3121 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3181 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3241 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3301 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3361 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3421 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3481 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3541 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3601 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     3661 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3721 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     3781 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
     3841 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
     3901 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     3961 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     4021 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     4081 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     4141 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     4201 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     4261 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     4321 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     4381 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     4441 cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg
     4501 cttatcgaaa ttaatacgac tcactatagg gagacccaag ctggctagcg tttaaactta
     4561 agcttggtac cgagctcgga tccactagtc cagtgtggtg gaattcatgg tgagcaaggg
     4621 cgaggagctg atcaaggaga acatgagaag caagctgtac ctggaaggca gcgtgaacgg
     4681 ccaccagttc aagtgcaccc acgaagggga gggcaagccc tacgagggca agcagaccaa
     4741 caggatcaag gtggtggagg gaggccccct gccgttcgca ttcgacatcc tggccaccca
     4801 ctttatgtac gggagcaagg tgttcatcaa gtaccccgcc gacctccccg attattttaa
     4861 gcagtccttc cctgagggct tcacatggga gagagtcatg gtgttcgaag acgggggcgt
     4921 gctgaccgcc acccaggaca ccagcctcca ggacggcgag ctcatctaca acgtcaaggt
     4981 cagaggggtg aacttcccag ccaacggccc cgtgatgcag aagaaaacac tgggctggga
     5041 gcccagcacc gagaccatgt accccgctga cggcggcctg gaaggcagat gcgacaaggc
     5101 cctgaagctc gtgggcgggg gccacctgca cgtcaacttc aagaccacat acaagtccaa
     5161 gaaacccgtg aagatgcccg gcgtccacta cgtggaccgc agactggaaa gaatcaagga
     5221 ggccgacaac gagacctacg tcgagcagta cgagcacgct gtggccagat actccaacct
     5281 gggcggaggc ttcacactcg aagatttcgt tggggactgg cgacagacag ccggctacaa
     5341 cctggaccaa gtccttgaac agggaggtgt gtccagtttg tttcagaatc tcggggtgtc
     5401 cgtaactccg atccaaagga ttgtcctgag cggtgaaaat gggctgaaga tcgacatcca
     5461 tgtcatcatc ccgtatgaag gtctgagcgg cgaccaaatg ggccagatcg aaaaaatttt
     5521 taaggtggtg taccctgtgg atgatcatca ctttaaggtg atcctgcact atggcacact
     5581 ggtaatcgac ggggttacgc cgaacatgat cgactatttc ggacggccgt atgaaggcat
     5641 cgccgtgttc gacggcaaaa agatcactgt aacagggacc ctgtggaacg gcaacaaaat
     5701 tatcgacgag cgcctgatca accccgacgg ctccctgctg ttccgagtaa ccatcaacgg
     5761 agtgaccggc tggcggctgt gcgaacgcat tctggcgagg cacgagctga tcaaggagaa
     5821 catgagaagc aagctgtacc tggaaggcag cgtgaacggc caccagttca agtgcaccca
     5881 cgaaggggag ggcaagccct acgagggcaa gcagaccaac aggatcaagg tggtggaggg
     5941 aggccccctg ccgttcgcat tcgacatcct ggccacccac tttatgtacg ggagcaaggt
     6001 gttcatcaag taccccgccg acctccccga ttattttaag cagtccttcc ctgagggctt
     6061 cacatgggag agagtcatgg tgttcgaaga cgggggcgtg ctgaccgcca cccaggacac
     6121 cagcctccag gacggcgagc tcatctacaa cgtcaaggtc agaggggtga acttcccagc
     6181 caacggcccc gtgatgcaga agaaaacact gggctgggag cccagcaccg agaccatgta
     6241 ccccgctgac ggcggcctgg aaggcagatg cgacaaggcc ctgaagctcg tgggcggggg
     6301 ccacctgcac gtcaacttca agaccacata caagtccaag aaacccgtga agatgcccgg
     6361 cgtccactac gtggaccgca gactggaaag aatcaaggag gccgacaacg agacctacgt
     6421 cgagcagtac gagcacgctg tggccagata ctccaacctg ggcggaggca tggacgagct
     6481 gtacaaggcg gccgctgagg gcagaggaag tcttctaaca tgcggtgacg tggaggagaa
     6541 tcccggccct tccgggatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     6601 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     6661 caccgtcgat ccggaccgcc acctcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     6721 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     6781 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     6841 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     6901 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     6961 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     7021 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     7081 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     7141 caagcccggt gcctgaaccc ctctccctcc ccccccccta acgttactgg ccgaagccgc
     7201 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     7261 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     7321 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg
     7381 gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
     7441 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
     7501 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
     7561 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
     7621 gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta
     7681 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atgtgatggt
     7741 tgagatgttg ccaaccgtag ctgtgctcgt tttggctgtc agcgtagtcg ctaaagataa
     7801 tacaacactt caggagttcg caacatctaa gggagaagag ttgtttactg gcgtagtacc
     7861 cattcttgtg gaacttgacg gcgatgttaa tggtcataag ttctctgtgt ctggtgaggg
     7921 cgagggggat gctacatatg gtaaattgac cctcaaattt atttgtacaa ccgggaagct
     7981 ccctgtaccc tggcctactc tcgtgaccac cctcacctac ggggtacaat gtttttccag
     8041 ataccccgat cacatgaagc gccacgattt cttcaaaagt gccatgcccg agggttacgt
     8101 ccaggaacgc acaatatttt tcaaggatga tggaaattac aaaacccgcg ccgaggttaa
     8161 gttcgagggg gataccctcg ttaaccggat tgaactgaag ggtatcgact ttaaggagga
     8221 cggtaatata ttggggcata aattggaata caactataac gaccaccagg tgtatatcat
     8281 ggccgataaa cagaagaacg gcatcaaagc aaactttaaa attaggcata acatagagga
     8341 tggtggcgtt cagcttgctg atcactatca acagaatacc cctataggtg atggacccgt
     8401 gctcctccca gataaccatt acttgtttac aacttctaca ctctctaagg accccaacga
     8461 aaagagagac cacatggtgc tcttggagtt cgtaactgcc gccggaatta cacatggtat
     8521 ggacgagctc tacaaggata tttccggggg ggtctttaca cttgaagatt tcgtcgggga
     8581 ttggagacaa actgccggtt ataatcttga tcaggtgctt gagcaagggg ggctgagctc
     8641 cctttttcaa aagctgggtg tatctgtcac ccctatccaa aagattgtgc ttagcggtga
     8701 aaatggtctc aaaatcgaca tccacgtgat catcccatac gagggtctta gtggatacca
     8761 gatggggcag atagagaaga ttttcaaagt tgtttaccct gtcgacgacc accatttcaa
     8821 agtaattctg cactacggaa ccctcgtcat agacggggtc acacccaaca tgatagatta
     8881 ctttgggcga ccctatgaag ggatagcagt attcgacggc aaaaagataa ccgtgacagg
     8941 caccctgtgg aatggaaaca aaattataga tgagcgcctg atcaaccctg acggttcttt
     9001 gttgttcagg gttacaatca atggtgtcac aggctggagg ctctgtgaga acatcctggc
     9061 tcagttggag aatgggggga tctccctctt ggtgcaaaat acatcatgga tgttgcttct
     9121 tcttcttagc ttgagcctct tgcaggcatt ggatttcatc agtctctaa
//
