LOCUS       sequence-345350-       12966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The lentiviral vector for Booster-PKA, a Red-Shifted genetically 
            encoded FRET biosensor for monitoring PKA activity..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12966)
  AUTHORS   Watabe T, Terai K, Sumiyama K, Matsuda M
  TITLE     Booster, a Red-Shifted Genetically Encoded Forster Resonance Energy 
            Transfer (FRET) Biosensor Compatible with Cyan Fluorescent 
            Protein/Yellow Fluorescent Protein-Based FRET Biosensors and Blue 
            Light-Responsive Optogenetic Tools.
  JOURNAL   ACS Sens. 2020 Feb 26. doi: 10.1021/acssensors.9b01941.
  PUBMED    32101394
REFERENCE   2  (bases 1 to 12966)
  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: "ACS Sens. 
            2020 Feb 26. doi: 10.1021/acssensors.9b01941."
COMMENT     Sequence Label: pCSIIpuro-4493NES
FEATURES             Location/Qualifiers
     source          1..12966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1442..1675
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1860..1904
                     /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    2178..2295
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2351..3513
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2582..3504
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3461..3481
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3538..3547
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3544..4239
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     CDS             4675..5022
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             4771..5118
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             4867..5214
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             4963..5310
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             5059..5406
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             5413..6066
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     CDS             6112..6138
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     misc_feature    6241..6790
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6385..6402)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6612..6631
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6793..7392
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6793..6812)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7289..7309
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7427..8015
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7480..7500)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7898..7909)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8534..8714
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8740..8757)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8746..8970
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9016..9444
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9103..9122)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9313..9334
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9453..9473)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9458..9787
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9638..9773
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9700..9719
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9835..9882
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9901..10275
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    10405..10526
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10442..10461)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10496..10515
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10575..10591)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10575..10591)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10588..10610)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10599..10615
                     /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(10623..10629,10630..10647,10648..10653))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10668..10689
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10806..10823)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10977..11565)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11057..11076)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11736..12527,12528..12596))
                     /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     12359..12378
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12597..12701)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12776..12795)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagggc cgctgatctt cagacctgga ggaggagata
     1321 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
     1381 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
     1441 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
     1501 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
     1561 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
     1621 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
     1681 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
     1741 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
     1801 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
     1861 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
     1921 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
     1981 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
     2041 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag
     2101 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tctacaaatg
     2161 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2221 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2281 aaaattcaaa attttcgggt ttattacagg gacagcagaa attcactttg aattaattca
     2341 agcttcgtga ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
     2401 gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa
     2461 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
     2521 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     2581 ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt
     2641 gccttgaatt acttccacct ggctccagta cgtgattctt gatcccgagc tggagccagg
     2701 ggcgggcctt gcgctttagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg
     2761 cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat
     2821 aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat
     2881 agtcttgtaa atgcgggcca ggatctgcac actggtattt cggtttttgg ggccgcgggc
     2941 ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg
     3001 ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc
     3061 gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg
     3121 tgagcggaaa gatggccgct tcccggccct gctccagggg gctcaaaatg gaggacgcgg
     3181 cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaggggcctt tccgtcctca
     3241 gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc
     3301 tggagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt
     3361 ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc
     3421 ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt
     3481 caaagttttt ttcttccatt tcaggtgtcg tgaacacgct accggtccga attcgccgcc
     3541 accatggtga gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc
     3601 gtgaacaacc accacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
     3661 cagaccatga gaatcaaggc ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
     3721 gctaccagct tcatgtacgg cagcaaaacc ttcatcaacc acacccaggg catccccgac
     3781 ttctttaagc agtccttccc cgagggcttc acatgggaga gagtcaccac atacgaagac
     3841 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
     3901 gtcaagatca gaggggtgaa cttcccatcc aacggccctg tgatgcagaa gaaaacactc
     3961 ggctgggagg cctccaccga gaccctgtac cccgctgacg gcggcctgga aggcagagcc
     4021 gacatggccc tgaagctcgt gggcgggggc cacctgatct gcaacttgaa gaccacatac
     4081 agatccaaga aacccgctaa gaacctcaag atgcccggcg tctactatgt ggacagaaga
     4141 ctggaaagaa tcaaggaggc cgacaaagag acctacgtcg agcagcacga ggtggctgtg
     4201 gccagatact gcgacctccc tagcaaactg gggcacagac tcgagaagtt ttctcaagaa
     4261 cagatcggcg aaaacattgt gtgcagggtc atttgtacca cgggtcaaat tcccatccga
     4321 gatttgtcag ctgatatttc acaagtgctt aaggaaaaac gatccataaa gaaagtttgg
     4381 acatttggta gaaacccagc ctgtgactat catttaggaa acatttcaag actgtcaaat
     4441 aagcatttcc aaatactact aggagaagac ggtaaccttt tattgaatga catttccact
     4501 aatgggacct ggttaaatgg gcaaaaagtc gagaagaaca gcaatcagtt actgtcccaa
     4561 ggtgatgaaa taaccgttgg tgtaggcgtg gaatcagata ttttatctct ggtcattttc
     4621 ataaacgaca aatttaagca gtgcctggag cagaacaaag ttgatcgcgg taccagtgct
     4681 ggtggtagtg ctggtggtag tgctggtggt agtgctggtg gtagtgctgg tggttccggc
     4741 agtgctggtg gtagtgctgg tggtagtacc agtgctggtg gtagtgctgg tggtagtgct
     4801 ggtggtagtg ctggtggtag tgctggtggt tccggcagtg ctggtggtag tgctggtggt
     4861 agtaccagtg ctggtggtag tgctggtggt agtgctggtg gtagtgctgg tggtagtgct
     4921 ggtggttccg gcagtgctgg tggtagtgct ggtggtagta ccagtgctgg tggtagtgct
     4981 ggtggtagtg ctggtggtag tgctggtggt agtgctggtg gttccggcag tgctggtggt
     5041 agtgctggtg gtagtaccag tgctggtggt agtgctggtg gtagtgctgg tggtagtgct
     5101 ggtggtagtg ctggtggttc cggcagtgct ggtggtagtg ctggtggtag taccagtgct
     5161 ggtggtagtg ctggtggtag tgctggtggt agtgctggtg gtagtgctgg tggttccggc
     5221 agtgctggtg gtagtgctgg tggtagtacc agtgctggtg gtagtgctgg tggtagtgct
     5281 ggtggtagtg ctggtggtag tgctggtggt tccggcagtg ctggtggtag tgctggtggt
     5341 agtaccagtg ctggtggtag tgctggtggt agtgctggtg gtagtgctgg tggtagtgct
     5401 ggtggttccg gaatggtgag tgtgattaaa ccagagatga agatgaggta ctacatggac
     5461 ggctccgtca atgggcatga gttcacaatt gaaggtgaag gcacaggcag accttacgag
     5521 ggacatcaag agatgacact acgcgtcaca atggccgagg gcgggccaat gcctttcgcg
     5581 tttgacttag tgtcacacgt gttctgttac ggccacagag tatttactaa atatccagaa
     5641 gagataccag actatttcaa acaagcattt cctgaaggcc tgtcatggga aaggtcgttg
     5701 gagttcgaag atggtgggtc cgcttcagtc agtgcgcata taagccttag aggaaacacc
     5761 ttctaccaca aatccaaatt tactggggtt aactttcctg ccgatggtcc tatcatgcaa
     5821 aaccaaagtg ttgattggga gccatcaacc gagaaaatta ctgccagcga cggagttctg
     5881 aagggtgatg ttacgatgta cctaaaactt gaaggaggcg gcaatcacaa atgccaattc
     5941 aagactactt acaaggcggc aaaagagatt cttgaaatgc caggagacca ttacatcggc
     6001 catcgcctcg tcaggaaaac cgaaggcaac attactgagc aggtagaaga tgcagtagct
     6061 cattccgcgg ccgcattgag gcgcgcgacg ctggttgact ctagactgca gctgcctcct
     6121 ctggaacgcc tgactctgga ttaagtcgag ggcggcggcc gcggctctag aggatccgtt
     6181 aactaactta agctagcaac ggtttccctc tagcgggatc aattccgccc ccccccccta
     6241 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     6301 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     6361 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     6421 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     6481 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     6541 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     6601 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     6661 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     6721 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     6781 cacgataata ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     6841 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     6901 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     6961 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     7021 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     7081 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     7141 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     7201 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     7261 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     7321 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     7381 cccggtgcct gactagagtt aacatcgagg gatcaagctt atcgataatc aacctctgga
     7441 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     7501 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     7561 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     7621 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     7681 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     7741 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     7801 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     7861 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     7921 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     7981 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgag acctagaaaa
     8041 acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga
     8101 agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat
     8161 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     8221 gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg
     8281 ctacttccct gattggcaga actacacacc agggccaggg atcagatatc cactgacctt
     8341 tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg
     8401 agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
     8461 agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca
     8521 tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     8581 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     8641 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     8701 gaaaatctct agcagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     8761 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     8821 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     8881 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     8941 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     9001 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     9061 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     9121 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     9181 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     9241 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     9301 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     9361 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     9421 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     9481 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     9541 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     9601 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     9661 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     9721 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     9781 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
     9841 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     9901 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc
     9961 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt
    10021 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac
    10081 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag
    10141 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
    10201 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc
    10261 gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg
    10321 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
    10381 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
    10441 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
    10501 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
    10561 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
    10621 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
    10681 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
    10741 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
    10801 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
    10861 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
    10921 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
    10981 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
    11041 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
    11101 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
    11161 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
    11221 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
    11281 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
    11341 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
    11401 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
    11461 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
    11521 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
    11581 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
    11641 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
    11701 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
    11761 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
    11821 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
    11881 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
    11941 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
    12001 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
    12061 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
    12121 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
    12181 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
    12241 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
    12301 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
    12361 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
    12421 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
    12481 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
    12541 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
    12601 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
    12661 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
    12721 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
    12781 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
    12841 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
    12901 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
    12961 cgcgtt
//
