LOCUS       sequence-408625-       11602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GPCR/G protein BRET biosensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11602)
  AUTHORS   Janicot R, Maziarz M, Park JC, Zhao J, Luebbers A, Green E, 
            Philibert CE, Zhang H, Layne MD, Wu JC, Garcia-Marcos M
  TITLE     Direct interrogation of context-dependent GPCR activity with a 
            universal biosensor platform.
  JOURNAL   Cell. 2024 Feb 16:S0092-8674(24)00065-5. doi: 
            10.1016/j.cell.2024.01.028.
  PUBMED    38412860
REFERENCE   2  (bases 1 to 11602)
  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: "Cell. 2024 
            Feb 16:S0092-8674(24)00065-5. doi: 10.1016/j.cell.2024.01.028."
COMMENT     Sequence Label: pLVX-hSyn-Galpha-i3-IRESWT-Gbetagamma ONE-GO
FEATURES             Location/Qualifiers
     source          1..11602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    189..304
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        346..793
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     misc_feature    1895..2468
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1918..2470
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2062..2079)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2289..2308
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2470..2490
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             3097..3609
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             3619..3672
                     /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             4693..4746
                     /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             4753..5487
                     /codon_start=1
                     /product="circularly permuted form of the Venus yellow 
                     fluorescent protein (Nagai et al., 2004)"
                     /label=cp173Venus
                     /note="efficient FRET acceptor for CFP"
                     /translation="MDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRD
                     HMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYITADKQKNGIKANFKIRHNIE"
     primer_bind     4890..4911
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5014..5035)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5275..5294)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    5732..6320
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5785..5805)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6203..6214)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6527..7160
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7288..7304)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7288..7304)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7301..7323)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7312..7328
                     /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(7336..7342,7343..7360,7361..7366))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7381..7402
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7519..7536)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7690..8278)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7770..7789)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8449..9240,9241..9309))
                     /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     9072..9091
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9310..9414)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    9462..9596
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9499..9518)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9553..9572
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             9764..10397
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10444..10569
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11066..11299
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11484..11528
                     /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"
ORIGIN
        1 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
       61 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
      121 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg
      181 gtatcgcctt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag
      241 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
      301 attttcgggt ttattacagg gacagcagag atccagttta tcgatagtgc aagtgggttt
      361 taggaccagg atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga
      421 caagcaccca acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa
      481 caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc
      541 cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc
      601 cggtcccccg caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc
      661 agccggaccg caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg
      721 cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg
      781 cctgagagcg cagctcgaga ctagttctag agcggccgca tgggctgcac gttgagcgcc
      841 gaggacaagg cggcggtgga gcggagtaag atgatcgacc gcaacttgcg ggaggacgga
      901 gagaaagcgg ccaaagaagt gaagctgctg ctgctcggcg ctggagaatc tggtaaaagt
      961 actattgtga aacagatgaa aatcattcat gaggatggct attccgagga cgagtgtaag
     1021 cagtataaag ttgtcgtcta cagcaatacc attcagtcca tcattgcaat cataagagcc
     1081 atgggacggt tgaagattga ttttggggaa gctgccagag cggatgatgc ccgacagtta
     1141 tttgttttag ctggcagtgc tgaggaagga gtcatgactt cagaactagc aggcgtgatt
     1201 aaacgtttat ggcgagatgg cggggtgcag gcatgcttca gcaggtccag ggaatatcag
     1261 ctcaatgatt ctgcttcata ttacctaaat gatttggata gaatatccca gaccaactac
     1321 attccaactc agcaagatgt tcttcggacg agagtgaaga ctacaggcat tgtggagacc
     1381 cacttcacct tcaaggagct ctacttcaaa atgtttgatg taggtggcca aagatccgaa
     1441 cggaaaaagt ggatccactg ttttgaggga gtgacagcaa ttatcttttg tgtggctctc
     1501 agtgattacg accttgttct ggctgaggac gaggaaatga accgaatgca tgaaagcatg
     1561 aaattgtttg acagcatttg taacaacaaa tggtttacag acacttcaat cattctcttc
     1621 cttaataaga aagacctttt tgaggaaaaa ataaagagga gtccattaac aatctgttat
     1681 ccagaataca caggttccaa tacgtacgaa gaggcagctg cttacatcca gtgccagttt
     1741 gaagatctga accgaagaaa ggacaccaag gaggtctaca ctcactttac ctgtgccact
     1801 gacaccaaaa acgtgcagtt tgtttttgat gctgttacag atgtcatcat taaaaacaac
     1861 ttaaaggaat gtgggcttta ctgagcggcc gccgcccctc tccctccccc ccccctaacg
     1921 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     1981 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     2041 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     2101 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     2161 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     2221 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     2281 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     2341 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     2401 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     2461 gatgataata tggggagtag caagagcaag acgtcgaatt ccggaatcaa gcttcttgat
     2521 tgcgaccaag aactctacaa gaacttccct ctggtgatct ctgagcgctg gcagcaggaa
     2581 gtggcggaaa cagtttatga agcagtaaat gcagacacgg ataaaatcga ggccaggaag
     2641 agagctaaaa ataagcagct tggccacgaa gaagattacg ccctgggaag agactgcatc
     2701 gtgcacgggt acatgctgaa gctggggaac cctttcctga cccagtggca gcgccgctat
     2761 ttttacctct ttccgaacag acttgagtgg agaggagaag gcgagtcgcg acaaagttta
     2821 ctgacaatgg aacagattgt gtccgtggaa gaaactcaga ttaaagacaa aaagtgcatt
     2881 ttgttgagaa taaaaggagg gaagcagttc gttttgcagt gtgagagtga cccagagttt
     2941 gtgcagtgga agaaagagct gacggagaca ttcatggagg cccagcggct gctacggcga
     3001 gcccccaagt tcctcaacaa atcccgctca gccgtcgtgg aactctcaaa gcctcccctc
     3061 tgccatagga acagcaacgg cctcggaggt ggatccatgg tcttcacact cgaagatttc
     3121 gttggggact ggcgacagac agccggctac aacctggacc aagtccttga acagggaggt
     3181 gtgtccagtt tgtttcagaa tctcggggtg tccgtaactc cgatccaaag gattgtcctg
     3241 agcggtgaaa atgggctgaa gatcgacatc catgtcatca tcccgtatga aggtctgagc
     3301 ggcgaccaaa tgggccagat cgaaaaaatt tttaaggtgg tgtaccctgt ggatgatcat
     3361 cactttaagg tgatcctgca ctatggcaca ctggtaatcg acggggttac gccgaacatg
     3421 atcgactatt tcggacggcc gtatgaaggc atcgccgtgt tcgacggcaa aaagatcact
     3481 gtaacaggga ccctgtggaa cggcaacaaa attatcgacg agcgcctgat caaccccgac
     3541 ggctccctgc tgttccgagt aaccatcaac ggagtgaccg gctggcggct gtgcgaacgc
     3601 attctggcgg gatcaggaga aggtagagga agtctactaa catgcgggga cgtggaggaa
     3661 aatcccggcc caatggggga gcttgaccag ttacggcagg aggccgagca acttaagaac
     3721 cagattcgag acgccaggaa agcatgtgca gatgcaactc tctctcagat cacaaacaac
     3781 atcgacccag tgggaagaat ccaaatgcgc acgaggagga cactgcgggg gcacctggcc
     3841 aagatctacg ccatgcactg gggcacagac tccaggcttc tcgtcagtgc ctcgcaggat
     3901 ggtaaactta tcatctggga cagctacacc accaacaagg tccacgccat ccctctgcgc
     3961 tcctcctggg tcatgacctg tgcatatgcc ccttctggga actatgtggc ctgcggtggc
     4021 ctggataaca tttgctccat ttacaatctg aaaactcgtg aggggaacgt gcgcgtgagt
     4081 cgtgagctgg caggacacac aggttacctg tcctgctgcc gattcctgga tgacaatcag
     4141 atcgtcacca gctctggaga caccacgtgt gccctgtggg acatcgagac cggccagcag
     4201 acgaccacgt ttaccggaca cactggagat gtcatgagcc tttctcttgc tcctgacacc
     4261 agactgttcg tctctggtgc ttgtgatgct tcagccaaac tctgggatgt gcgagaaggc
     4321 atgtgccggc agaccttcac tggccacgag tctgacatca atgccatttg cttctttcca
     4381 aatggcaatg catttgccac tggctcagac gacgccacct gcaggctgtt tgaccttcgt
     4441 gctgaccagg agctcatgac ttactcccat gacaacatca tctgcgggat cacctctgtc
     4501 tccttctcca agagcgggcg cctcctcctt gctgggtacg acgacttcaa ctgcaacgtc
     4561 tgggatgcac tcaaagccga ccgggcaggt gtcttggctg ggcatgacaa ccgcgtcagc
     4621 tgcctgggcg tgactgacga tggcatggct gtggcgacag ggtcctggga tagcttcctc
     4681 aagatctgga acgagggcag aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc
     4741 ggccctaccg gtatggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc
     4801 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg
     4861 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     4921 gggatcactc tcggcatgga cgagctgtac aagggtggca gcggtggcat ggtgagcaag
     4981 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     5041 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     5101 ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     5161 ctgggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     5221 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     5281 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     5341 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     5401 aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc
     5461 aacttcaaga tccgccacaa catcgagtcc ggactcagat ctatggccag caacaacacc
     5521 gccagcatag cacaagccag gaagctggta gagcagctta agatggaagc caatatcgac
     5581 aggataaagg tgtccaaggc agctgcagat ttgatggcct actgtgaagc acatgccaag
     5641 gaagaccccc tcctgacccc tgttccggct tcagaaaacc cgtttaggga gaagaagttt
     5701 ttctgtgcca tcctttaaac gcgtctggaa caatcaacct ctggattaca aaatttgtga
     5761 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     5821 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5881 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     5941 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     6001 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     6061 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     6121 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     6181 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     6241 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     6301 cctttgggcc gcctccccgc ctggaattaa ttctgcagtc gagacctaga aaaacatgga
     6361 gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa
     6421 gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac
     6481 aaggcagctg tagatcttag ccacttttta aaagaaaaga ggggactgga agggctaatt
     6541 cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc
     6601 cctgattagc agaactacac accagggcca ggggtcagat atccactgac ctttggatgg
     6661 tgctacaagc tagtaccagt tgagccagat aaggtagaag aggccaataa aggagagaac
     6721 accagcttgt tacaccctgt gagcctgcat gggatggatg acccggagag agaagtgtta
     6781 gagtggaggt ttgacagccg cctagcattt catcacgtgg cccgagagct gcatccggag
     6841 tacttcaaga actgctgata tcgagcttgc tacaagggac tttccgctgg ggactttcca
     6901 gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatcctg catataagca
     6961 gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     7021 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     7081 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     7141 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     7201 tgcaaagaaa tgaatatcag agagtgagag gccttgacat tgctagcgtt taccgtcgac
     7261 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     7321 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     7381 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7441 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7501 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7561 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7621 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7681 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7741 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7801 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7861 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7921 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7981 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     8041 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     8101 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     8161 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     8221 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     8281 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     8341 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     8401 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8461 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8521 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8581 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8641 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8701 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8761 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8821 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8881 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8941 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     9001 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     9061 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     9121 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     9181 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     9241 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     9301 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     9361 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     9421 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat caacttgttt attgcagctt
     9481 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     9541 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaact
     9601 ggataactca agctaaccaa aatcatccca aacttcccac cccataccct attaccactg
     9661 ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta ttttcatttt
     9721 aaagaaattg tatttgttaa atatgtacta caaacttagt agttggaagg gctaattcac
     9781 tcccaaagaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct
     9841 gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc
     9901 tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc
     9961 agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag
    10021 tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac
    10081 ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg
    10141 aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct
    10201 gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
    10261 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
    10321 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
    10381 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag
    10441 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
    10501 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
    10561 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg
    10621 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa
    10681 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga
    10741 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta
    10801 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac
    10861 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cggccgctga
    10921 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
    10981 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
    11041 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
    11101 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
    11161 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
    11221 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
    11281 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
    11341 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
    11401 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
    11461 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
    11521 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
    11581 tgatagtagg aggcttggta gg
//
