LOCUS       sequence-405587-       11686 bp    DNA     circular SYN 14-APR-2025
DEFINITION  GPCR/G protein BRET biosensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11686)
  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 11686)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-s-IRESWT-Gbetagamma ONE-GO
FEATURES             Location/Qualifiers
     source          1..11686
                     /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    1979..2552
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2002..2554
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2146..2163)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2373..2392
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2554..2574
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             3181..3693
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     CDS             3703..3756
                     /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             4777..4830
                     /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             4837..5571
                     /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     4974..4995
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5098..5119)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5359..5378)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    5816..6404
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5869..5889)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6287..6298)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6611..7244
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7372..7388)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7372..7388)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7385..7407)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7396..7412
                     /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(7420..7426,7427..7444,7445..7450))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7465..7486
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7603..7620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7774..8362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7854..7873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8533..9324,9325..9393))
                     /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     9156..9175
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9394..9498)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    9546..9680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9583..9602)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9637..9656
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             9848..10481
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    10528..10653
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11150..11383
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11568..11612
                     /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 agcggccgcg gatccatggg ctgcctcggg
      841 aacagtaaga ccgaggacca gcgcaacgag gagaaggcgc agcgtgaggc caacaaaaag
      901 atcgagaagc agctgcagaa ggacaagcag gtctaccggg ccacgcaccg cctgctgctg
      961 ctgggtgctg gagaatctgg taaaagcacc attgtgaagc agatgaggat cctgcatgtt
     1021 aatgggttta atggagacag tgagaaggca accaaagtgc aggacatcaa aaacaacctg
     1081 aaagaggcga ttgaaaccat tgtggccgcc atgagcaacc tggtgccccc cgtggagctg
     1141 gccaaccccg agaaccagtt cagagtggac tacattctga gtgtgatgaa cgtgcctgac
     1201 tttgacttcc ctcccgaatt ctatgagcat gccaaggctc tgtgggagga tgaaggagtg
     1261 cgtgcctgct acgaacgctc caacgagtac cagctgattg actgtgccca gtacttcctg
     1321 gacaagatcg acgtgatcaa gcaggctgac tatgtgccga gcgatcagga cctgcttcgc
     1381 tgccgtgtcc tgacttctgg aatctttgag accaagttcc aggtggacaa agtcaacttc
     1441 cacatgtttg acgtgggtgg ccagcgcgat gaacgccgca agtggatcca gtgcttcaac
     1501 gatgtgactg ccatcatctt cgtggtggcc agcagcagct acaacatggt catccgggag
     1561 gacaaccaga ccaaccgcct gcaggaggct ctgaacctct tcaagagcat ctggaacaac
     1621 agatggctgc gcaccatctc tgtgatcctg ttcctcaaca agcaagatct gctcgctgag
     1681 aaagtccttg ctgggaaatc gaagattgag gactactttc cagaatttgc tcgctacact
     1741 actcctgagg atgctactcc cgagcccgga gaggacccac gcgtgacccg ggccaagtac
     1801 ttcattcgag atgagtttct gaggatcagc actgccagtg gagatgggcg tcactactgc
     1861 taccctcatt tcacctgcgc tgtggacact gagaacatcc gccgtgtgtt caacgactgc
     1921 cgtgacatca ttcagcgcat gcaccttcgt cagtacgagc tgctataagc ggccgccgcc
     1981 cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     2041 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     2101 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     2161 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     2221 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     2281 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     2341 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     2401 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca
     2461 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     2521 acgtggtttt cctttgaaaa acacgatgat aatatgggga gtagcaagag caagacgtcg
     2581 aattccggaa tcaagcttct tgattgcgac caagaactct acaagaactt ccctctggtg
     2641 atctctgagc gctggcagca ggaagtggcg gaaacagttt atgaagcagt aaatgcagac
     2701 acggataaaa tcgaggccag gaagagagct aaaaataagc agcttggcca cgaagaagat
     2761 tacgccctgg gaagagactg catcgtgcac gggtacatgc tgaagctggg gaaccctttc
     2821 ctgacccagt ggcagcgccg ctatttttac ctctttccga acagacttga gtggagagga
     2881 gaaggcgagt cgcgacaaag tttactgaca atggaacaga ttgtgtccgt ggaagaaact
     2941 cagattaaag acaaaaagtg cattttgttg agaataaaag gagggaagca gttcgttttg
     3001 cagtgtgaga gtgacccaga gtttgtgcag tggaagaaag agctgacgga gacattcatg
     3061 gaggcccagc ggctgctacg gcgagccccc aagttcctca acaaatcccg ctcagccgtc
     3121 gtggaactct caaagcctcc cctctgccat aggaacagca acggcctcgg aggtggatcc
     3181 atggtcttca cactcgaaga tttcgttggg gactggcgac agacagccgg ctacaacctg
     3241 gaccaagtcc ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg ggtgtccgta
     3301 actccgatcc aaaggattgt cctgagcggt gaaaatgggc tgaagatcga catccatgtc
     3361 atcatcccgt atgaaggtct gagcggcgac caaatgggcc agatcgaaaa aatttttaag
     3421 gtggtgtacc ctgtggatga tcatcacttt aaggtgatcc tgcactatgg cacactggta
     3481 atcgacgggg ttacgccgaa catgatcgac tatttcggac ggccgtatga aggcatcgcc
     3541 gtgttcgacg gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa caaaattatc
     3601 gacgagcgcc tgatcaaccc cgacggctcc ctgctgttcc gagtaaccat caacggagtg
     3661 accggctggc ggctgtgcga acgcattctg gcgggatcag gagaaggtag aggaagtcta
     3721 ctaacatgcg gggacgtgga ggaaaatccc ggcccaatgg gggagcttga ccagttacgg
     3781 caggaggccg agcaacttaa gaaccagatt cgagacgcca ggaaagcatg tgcagatgca
     3841 actctctctc agatcacaaa caacatcgac ccagtgggaa gaatccaaat gcgcacgagg
     3901 aggacactgc gggggcacct ggccaagatc tacgccatgc actggggcac agactccagg
     3961 cttctcgtca gtgcctcgca ggatggtaaa cttatcatct gggacagcta caccaccaac
     4021 aaggtccacg ccatccctct gcgctcctcc tgggtcatga cctgtgcata tgccccttct
     4081 gggaactatg tggcctgcgg tggcctggat aacatttgct ccatttacaa tctgaaaact
     4141 cgtgagggga acgtgcgcgt gagtcgtgag ctggcaggac acacaggtta cctgtcctgc
     4201 tgccgattcc tggatgacaa tcagatcgtc accagctctg gagacaccac gtgtgccctg
     4261 tgggacatcg agaccggcca gcagacgacc acgtttaccg gacacactgg agatgtcatg
     4321 agcctttctc ttgctcctga caccagactg ttcgtctctg gtgcttgtga tgcttcagcc
     4381 aaactctggg atgtgcgaga aggcatgtgc cggcagacct tcactggcca cgagtctgac
     4441 atcaatgcca tttgcttctt tccaaatggc aatgcatttg ccactggctc agacgacgcc
     4501 acctgcaggc tgtttgacct tcgtgctgac caggagctca tgacttactc ccatgacaac
     4561 atcatctgcg ggatcacctc tgtctccttc tccaagagcg ggcgcctcct ccttgctggg
     4621 tacgacgact tcaactgcaa cgtctgggat gcactcaaag ccgaccgggc aggtgtcttg
     4681 gctgggcatg acaaccgcgt cagctgcctg ggcgtgactg acgatggcat ggctgtggcg
     4741 acagggtcct gggatagctt cctcaagatc tggaacgagg gcagaggaag tcttctaaca
     4801 tgcggtgacg tggaggagaa tcccggccct accggtatgg acggcggcgt gcagctcgcc
     4861 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     4921 tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     4981 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggt
     5041 ggcagcggtg gcatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     5101 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     5161 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg
     5221 ccctggccca ccctcgtgac caccctgggc tacggcctgc agtgcttcgc ccgctacccc
     5281 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     5341 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     5401 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     5461 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     5521 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga gtccggactc
     5581 agatctatgg ccagcaacaa caccgccagc atagcacaag ccaggaagct ggtagagcag
     5641 cttaagatgg aagccaatat cgacaggata aaggtgtcca aggcagctgc agatttgatg
     5701 gcctactgtg aagcacatgc caaggaagac cccctcctga cccctgttcc ggcttcagaa
     5761 aacccgttta gggagaagaa gtttttctgt gccatccttt aaacgcgtct ggaacaatca
     5821 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     5881 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     5941 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     6001 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     6061 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     6121 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     6181 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     6241 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     6301 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     6361 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc
     6421 agtcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct
     6481 gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag
     6541 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     6601 aagaggggac tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg
     6661 atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc
     6721 agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta
     6781 gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg
     6841 gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac
     6901 gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag
     6961 ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga
     7021 gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga
     7081 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     7141 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     7201 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     7261 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg
     7321 acattgctag cgtttaccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     7381 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     7441 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     7501 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     7561 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     7621 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     7681 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7741 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7801 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7861 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7921 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7981 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     8041 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     8101 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     8161 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     8221 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     8281 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     8341 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     8401 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     8461 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     8521 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     8581 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     8641 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8701 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8761 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8821 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8881 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8941 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     9001 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     9061 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     9121 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     9181 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     9241 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     9301 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     9361 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     9421 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     9481 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     9541 agatcaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     9601 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     9661 tatcttatca tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc
     9721 ccaccccata ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg
     9781 attcctctga attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact
     9841 tagtagttgg aagggctaat tcactcccaa agaagacaag atatccttga tctgtggatc
     9901 taccacacac aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga
     9961 tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa
    10021 gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat
    10081 gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg
    10141 gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga
    10201 ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc
    10261 ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca
    10321 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
    10381 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
    10441 atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac
    10501 ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg
    10561 cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga
    10621 ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg
    10681 cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata
    10741 gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca
    10801 gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa
    10861 cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata
    10921 aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc
    10981 gcacagcaag cggccggccg ctgatcttca gacctggagg aggagatatg agggacaatt
    11041 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
    11101 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
    11161 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg
    11221 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
    11281 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
    11341 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
    11401 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
    11461 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca
    11521 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
    11581 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc
    11641 tgtggtatat aaaattattc ataatgatag taggaggctt ggtagg
//
