pLVX-CMV-Galpha-i2 ONE-GO

LOCUS       pLVX-CMV-Galpha-i2 ONE-GO        9925 bp DNA     circular SYN 29-AUG-2024
DEFINITION  GPCR/G protein BRET biosensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9925)
  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 9925)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 29, 2024 from SnapGene Server 7.0.3
            https://www.biofeng.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."
FEATURES             Location/Qualifiers
     source          1..9925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     312..333
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1118..1704
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1285..1302)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1512..1531
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1715..1735
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSK"
     CDS             1841..2356
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     misc_feature    2370..2958
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2423..2443)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2841..2852)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3165..3798
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(3926..3942)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3926..3942)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3939..3961)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3950..3966
                     /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(3974..4004)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4019..4040
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4157..4174)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4328..4916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4408..4427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5087..5947)
                     /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     5710..5729
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5948..6052)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    6100..6234
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6137..6156)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6191..6210
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             6402..7035
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    7082..7207
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7704..7937
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8122..8166
                     /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"
     misc_feature    8429..8544
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     enhancer        8602..8905
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8906..9109
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9060..9080
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9106..9130
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(9584..9925,1..375)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(9629..9650)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(9890..9909)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
       61 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
      121 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
      181 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
      241 ttgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag
      301 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
      361 gacgagctgt acaagagcgg cgaggaacaa gggatgttgc cggaagatct gtcaggagtg
      421 attcggcggc tgtgggccga ccatggagtg caagcgtgct ttgggaggag ccgggaatac
      481 caactgaacg attccgccgc ttattacttg aacgaccttg agcgcattgc tcaaagcgat
      541 tatattccaa ctcaacaaga tgtactgaga acacgcgtga agacaacagg aatagtagag
      601 actcatttca cgtttaagga tctccacttt aaaatgttcg atgttggagg acaacgatct
      661 gagcgcaaga agtggataca ctgttttgag ggcgttaccg cgatcatttt ctgtgttgct
      721 ttgtctgcat acgacctcgt cttggcagag gatgaagaaa tgaatcgaat gcacgaatcc
      781 atgaagttgt ttgattctat atgcaacaat aagtggttta ctgataccag catcatactt
      841 ttcttgaata aaaaagactt gtttgaggag aagataacac agagcccact tacgatttgt
      901 ttcccggagt atacgggagc gaacaagtat gatgaagccg caagttatat ccaatccaag
      961 tttgaggacc tgaataagag aaaagacaca aaggagattt acacacactt cacctgtgcc
     1021 actgacacta aaaacgtcca atttgttttc gacgccgtta ccgatgtcat aataaaaaat
     1081 aacctcaaag actgtggtct tttttaagcg gccgccgccc ctctccctcc ccccccccta
     1141 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     1201 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     1261 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     1321 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     1381 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     1441 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     1501 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     1561 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     1621 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     1681 cacgatgata agcttgccac aacccgtacc aaagatgggg agtagcaaga gcaagacgtc
     1741 gaattccgga atcaagcttg gtggatcttc tcgaggttac taccatggta tttgggtggg
     1801 tgaagaaggt cgactttctc gaggtggcac cggtggatcc atggtcttca cactcgaaga
     1861 tttcgttggg gactggcgac agacagccgg ctacaacctg gaccaagtcc ttgaacaggg
     1921 aggtgtgtcc agtttgtttc agaatctcgg ggtgtccgta actccgatcc aaaggattgt
     1981 cctgagcggt gaaaatgggc tgaagatcga catccatgtc atcatcccgt atgaaggtct
     2041 gagcggcgac caaatgggcc agatcgaaaa aatttttaag gtggtgtacc ctgtggatga
     2101 tcatcacttt aaggtgatcc tgcactatgg cacactggta atcgacgggg ttacgccgaa
     2161 catgatcgac tatttcggac ggccgtatga aggcatcgcc gtgttcgacg gcaaaaagat
     2221 cactgtaaca gggaccctgt ggaacggcaa caaaattatc gacgagcgcc tgatcaaccc
     2281 cgacggctcc ctgctgttcc gagtaaccat caacggagtg accggctggc ggctgtgcga
     2341 acgcattctg gcgtaaacgc gtctggaaca atcaacctct ggattacaaa atttgtgaaa
     2401 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     2461 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2521 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     2581 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     2641 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     2701 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     2761 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     2821 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     2881 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     2941 tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa aacatggagc
     3001 aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga
     3061 ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa
     3121 ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag ggctaattca
     3181 ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
     3241 tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct ttggatggtg
     3301 ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag gagagaacac
     3361 cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtgttaga
     3421 gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc atccggagta
     3481 cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg actttccagg
     3541 gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca tataagcagc
     3601 tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     3661 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     3721 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     3781 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     3841 caaagaaatg aatatcagag agtgagaggc cttgacattg ctagcgttta ccgtcgacct
     3901 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3961 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4021 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4081 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4141 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4201 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4261 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4321 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4381 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4441 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4501 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4561 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4621 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4681 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4741 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4801 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4861 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4921 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4981 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5041 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5101 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5161 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5221 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5281 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5341 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5401 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5461 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5521 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5581 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5641 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5701 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5761 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5821 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5881 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5941 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6001 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6061 cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat tgcagcttat
     6121 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     6181 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaactgg
     6241 ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat taccactgcc
     6301 aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa
     6361 agaaattgta tttgttaaat atgtactaca aacttagtag ttggaagggc taattcactc
     6421 ccaaagaaga caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga
     6481 ttagcagaac tacacaccag ggccaggggt cagatatcca ctgacctttg gatggtgcta
     6541 caagctagta ccagttgagc cagataaggt agaagaggcc aataaaggag agaacaccag
     6601 cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag tgttagagtg
     6661 gaggtttgac agccgcctag catttcatca cgtggcccga gagctgcatc cggagtactt
     6721 caagaactgc tgatatcgag cttgctacaa gggactttcc gctggggact ttccagggag
     6781 gcgtggcctg ggcgggactg gggagtggcg agccctcaga tcctgcatat aagcagctgc
     6841 tttttgcctg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     6901 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     6961 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     7021 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     7081 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     7141 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     7201 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     7261 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     7321 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     7381 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     7441 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     7501 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg gccgctgatc
     7561 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
     7621 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
     7681 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
     7741 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt
     7801 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
     7861 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
     7921 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct
     7981 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc
     8041 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa
     8101 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca
     8161 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg
     8221 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga
     8281 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc
     8341 gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag atccattcga
     8401 ttagtgaacg gatctcgacg gtatcgcctt taaaagaaaa ggggggattg gggggtacag
     8461 tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag aattacaaaa
     8521 acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag atccagttta
     8581 tcgataagct tgggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     8641 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     8701 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     8761 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     8821 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     8881 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     8941 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     9001 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     9061 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
     9121 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccgact
     9181 ctactagagg atctatttcc ggtgaattcc tcgagactag ttctagagcg gccgcggatc
     9241 catgggatgc accgttagcg ccgaagacaa agcagcggcg gagcgaagca agatgattga
     9301 caaaaatttg agagaggatg gcgagaaggc ggctcgcgag gtcaaactgt tgcttctggg
     9361 tgccggagag agcggcaaga gcactattgt aaaacagatg aagatcattc atgaggatgg
     9421 ctactcagag gaggaatgtc ggcaataccg cgcagttgtg tatagtaata ctattcaatc
     9481 cattatggca attgtaaagg cgatgggcaa cctccagatc gactttgctg acccgcagag
     9541 ggcagacgac gcgaggcagc tgttcgcctt gagctgcgcc gctatggtga gcaagggcga
     9601 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     9661 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     9721 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg
     9781 ctacggcctg atgtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     9841 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     9901 ctacaagacc cgcgccgagg tgaag
//