LOCUS       sequence-405603-       10705 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 10705)
  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 10705)
  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-CMV-IRESWT-Gbetagamma ONE-GO
FEATURES             Location/Qualifiers
     source          1..10705
                     /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)"
     enhancer        362..665
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        666..869
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     820..840
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     866..890
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    998..1571
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1021..1573
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1165..1182)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1392..1411
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1573..1593
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             2200..2712
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             2722..2775
                     /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             3796..3849
                     /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             3856..4590
                     /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     3993..4014
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4117..4138)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4378..4397)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4835..5423
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4888..4908)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5306..5317)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5630..6263
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6391..6407)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6391..6407)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6404..6426)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6415..6431
                     /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(6439..6445,6446..6463,6464..6469))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6484..6505
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6622..6639)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6793..7381)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6873..6892)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7552..8343,8344..8412))
                     /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     8175..8194
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8413..8517)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    8565..8699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8602..8621)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8656..8675
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             8867..9500
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    9547..9672
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10169..10402
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10587..10631
                     /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 tcgataagct tgggagttcc
      361 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      421 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      481 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      541 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      601 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      661 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      721 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      781 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      841 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac
      901 gccatccacg ctgttttgac ctccatagaa gacaccgact ctactagagg atctatttcc
      961 ggtgaattcc tcgagactag ttctagagcg gccgccgccc ctctccctcc ccccccccta
     1021 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     1081 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     1141 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     1201 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     1261 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     1321 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     1381 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     1441 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     1501 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     1561 cacgatgata atatggggag tagcaagagc aagacgtcga attccggaat caagcttctt
     1621 gattgcgacc aagaactcta caagaacttc cctctggtga tctctgagcg ctggcagcag
     1681 gaagtggcgg aaacagttta tgaagcagta aatgcagaca cggataaaat cgaggccagg
     1741 aagagagcta aaaataagca gcttggccac gaagaagatt acgccctggg aagagactgc
     1801 atcgtgcacg ggtacatgct gaagctgggg aaccctttcc tgacccagtg gcagcgccgc
     1861 tatttttacc tctttccgaa cagacttgag tggagaggag aaggcgagtc gcgacaaagt
     1921 ttactgacaa tggaacagat tgtgtccgtg gaagaaactc agattaaaga caaaaagtgc
     1981 attttgttga gaataaaagg agggaagcag ttcgttttgc agtgtgagag tgacccagag
     2041 tttgtgcagt ggaagaaaga gctgacggag acattcatgg aggcccagcg gctgctacgg
     2101 cgagccccca agttcctcaa caaatcccgc tcagccgtcg tggaactctc aaagcctccc
     2161 ctctgccata ggaacagcaa cggcctcgga ggtggatcca tggtcttcac actcgaagat
     2221 ttcgttgggg actggcgaca gacagccggc tacaacctgg accaagtcct tgaacaggga
     2281 ggtgtgtcca gtttgtttca gaatctcggg gtgtccgtaa ctccgatcca aaggattgtc
     2341 ctgagcggtg aaaatgggct gaagatcgac atccatgtca tcatcccgta tgaaggtctg
     2401 agcggcgacc aaatgggcca gatcgaaaaa atttttaagg tggtgtaccc tgtggatgat
     2461 catcacttta aggtgatcct gcactatggc acactggtaa tcgacggggt tacgccgaac
     2521 atgatcgact atttcggacg gccgtatgaa ggcatcgccg tgttcgacgg caaaaagatc
     2581 actgtaacag ggaccctgtg gaacggcaac aaaattatcg acgagcgcct gatcaacccc
     2641 gacggctccc tgctgttccg agtaaccatc aacggagtga ccggctggcg gctgtgcgaa
     2701 cgcattctgg cgggatcagg agaaggtaga ggaagtctac taacatgcgg ggacgtggag
     2761 gaaaatcccg gcccaatggg ggagcttgac cagttacggc aggaggccga gcaacttaag
     2821 aaccagattc gagacgccag gaaagcatgt gcagatgcaa ctctctctca gatcacaaac
     2881 aacatcgacc cagtgggaag aatccaaatg cgcacgagga ggacactgcg ggggcacctg
     2941 gccaagatct acgccatgca ctggggcaca gactccaggc ttctcgtcag tgcctcgcag
     3001 gatggtaaac ttatcatctg ggacagctac accaccaaca aggtccacgc catccctctg
     3061 cgctcctcct gggtcatgac ctgtgcatat gccccttctg ggaactatgt ggcctgcggt
     3121 ggcctggata acatttgctc catttacaat ctgaaaactc gtgaggggaa cgtgcgcgtg
     3181 agtcgtgagc tggcaggaca cacaggttac ctgtcctgct gccgattcct ggatgacaat
     3241 cagatcgtca ccagctctgg agacaccacg tgtgccctgt gggacatcga gaccggccag
     3301 cagacgacca cgtttaccgg acacactgga gatgtcatga gcctttctct tgctcctgac
     3361 accagactgt tcgtctctgg tgcttgtgat gcttcagcca aactctggga tgtgcgagaa
     3421 ggcatgtgcc ggcagacctt cactggccac gagtctgaca tcaatgccat ttgcttcttt
     3481 ccaaatggca atgcatttgc cactggctca gacgacgcca cctgcaggct gtttgacctt
     3541 cgtgctgacc aggagctcat gacttactcc catgacaaca tcatctgcgg gatcacctct
     3601 gtctccttct ccaagagcgg gcgcctcctc cttgctgggt acgacgactt caactgcaac
     3661 gtctgggatg cactcaaagc cgaccgggca ggtgtcttgg ctgggcatga caaccgcgtc
     3721 agctgcctgg gcgtgactga cgatggcatg gctgtggcga cagggtcctg ggatagcttc
     3781 ctcaagatct ggaacgaggg cagaggaagt cttctaacat gcggtgacgt ggaggagaat
     3841 cccggcccta ccggtatgga cggcggcgtg cagctcgccg accactacca gcagaacacc
     3901 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc
     3961 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     4021 gccgggatca ctctcggcat ggacgagctg tacaagggtg gcagcggtgg catggtgagc
     4081 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     4141 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     4201 accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     4261 accctgggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     4321 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     4381 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     4441 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     4501 tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     4561 gccaacttca agatccgcca caacatcgag tccggactca gatctatggc cagcaacaac
     4621 accgccagca tagcacaagc caggaagctg gtagagcagc ttaagatgga agccaatatc
     4681 gacaggataa aggtgtccaa ggcagctgca gatttgatgg cctactgtga agcacatgcc
     4741 aaggaagacc ccctcctgac ccctgttccg gcttcagaaa acccgtttag ggagaagaag
     4801 tttttctgtg ccatccttta aacgcgtctg gaacaatcaa cctctggatt acaaaatttg
     4861 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     4921 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     4981 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     5041 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     5101 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     5161 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     5221 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     5281 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     5341 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     5401 ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct agaaaaacat
     5461 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
     5521 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
     5581 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact ggaagggcta
     5641 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac
     5701 ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact gacctttgga
     5761 tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa taaaggagag
     5821 aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagtg
     5881 ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga gctgcatccg
     5941 gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc tggggacttt
     6001 ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc ctgcatataa
     6061 gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct
     6121 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     6181 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     6241 gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata
     6301 acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc gtttaccgtc
     6361 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6421 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6481 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6541 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6601 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6661 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6721 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6781 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6841 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6901 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6961 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7021 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7081 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7141 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7201 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     7261 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7321 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7381 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7441 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7501 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7561 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7621 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7681 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7741 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7801 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7861 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7921 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7981 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8041 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8101 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8161 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8221 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8281 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8341 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8401 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8461 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8521 atttccccga aaagtgccac ctgacgtcga cggatcggga gatcaacttg tttattgcag
     8581 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     8641 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca
     8701 actggataac tcaagctaac caaaatcatc ccaaacttcc caccccatac cctattacca
     8761 ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa ttattttcat
     8821 tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagttgga agggctaatt
     8881 cactcccaaa gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc
     8941 cctgattagc agaactacac accagggcca ggggtcagat atccactgac ctttggatgg
     9001 tgctacaagc tagtaccagt tgagccagat aaggtagaag aggccaataa aggagagaac
     9061 accagcttgt tacaccctgt gagcctgcat gggatggatg acccggagag agaagtgtta
     9121 gagtggaggt ttgacagccg cctagcattt catcacgtgg cccgagagct gcatccggag
     9181 tacttcaaga actgctgata tcgagcttgc tacaagggac tttccgctgg ggactttcca
     9241 gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatcctg catataagca
     9301 gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     9361 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     9421 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     9481 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
     9541 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
     9601 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
     9661 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
     9721 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     9781 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     9841 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     9901 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
     9961 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccggccgc
    10021 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
    10081 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
    10141 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
    10201 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
    10261 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
    10321 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
    10381 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
    10441 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
    10501 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
    10561 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
    10621 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
    10681 taatgatagt aggaggcttg gtagg
//
