LOCUS       sequence-340876-       12972 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 12972)
  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 12972)
  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..12972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        616..814
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             832..1012
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1056..1181
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1678..1911
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2096..2140
                     /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    2414..2531
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2587..3749
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2818..3740
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3697..3717
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3778..3787
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3784..4479
                     /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             4915..5262
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             5011..5358
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             5107..5454
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             5203..5550
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             5299..5646
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             5653..6306
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     CDS             6352..6378
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     misc_feature    6481..7029
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6625..6642)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6852..6871
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7032..7631
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(7032..7051)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7528..7548
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7666..8254
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7719..7739)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8137..8148)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8773..8953
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8979..8996)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8985..9209
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9255..9683
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9342..9361)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9552..9573
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9692..9712)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9697..10027
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9878..10013
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9940..9959
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10075..10122
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10141..10515
                     /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    10645..10766
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10682..10701)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10736..10755
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10815..10831)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10815..10831)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10828..10850)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10839..10855
                     /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(10863..10869,10870..10887,10888..10893))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10908..10929
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11046..11063)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11217..11805)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11297..11316)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11976..12767,12768..12836))
                     /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     12599..12618
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12837..12941)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc
      841 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      901 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      961 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     1021 cgaacaggga cctgaaagcg aaagggaaac cagagctctc tcgacgcagg actcggcttg
     1081 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
     1141 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
     1201 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
     1261 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
     1321 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
     1381 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     1441 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     1501 agaccaccgc acagcaagcg gccggccgct gatcttcaga cctggaggag gagatatgag
     1561 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1621 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1681 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cctcaatgac
     1741 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1801 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1861 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1921 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1981 taaatctctg gaacagatct ggaatcacac gacctggatg gagtgggaca gagaaattaa
     2041 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     2101 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     2161 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     2221 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     2281 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     2341 aggtggagag agagacagag acagatccat tcgattagtg aacggatcta caaatggcag
     2401 tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
     2461 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa
     2521 ttcaaaattt tcgggtttat tacagggaca gcagaaattc actttgaatt aattcaagct
     2581 tcgtgaggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     2641 ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg
     2701 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     2761 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta
     2821 agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct
     2881 tgaattactt ccacctggct ccagtacgtg attcttgatc ccgagctgga gccaggggcg
     2941 ggccttgcgc tttaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
     3001 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
     3061 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
     3121 ttgtaaatgc gggccaggat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
     3181 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
     3241 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
     3301 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     3361 cggaaagatg gccgcttccc ggccctgctc cagggggctc aaaatggagg acgcggcgct
     3421 cgggagagcg ggcgggtgag tcacccacac aaaggaaagg ggcctttccg tcctcagccg
     3481 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctgga
     3541 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     3601 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     3661 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     3721 gtttttttct tccatttcag gtgtcgtgaa cacgctaccg gtctcgacga attcgccgcc
     3781 accatggtga gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc
     3841 gtgaacaacc accacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
     3901 cagaccatga gaatcaaggc ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
     3961 gctaccagct tcatgtacgg cagcaaaacc ttcatcaacc acacccaggg catccccgac
     4021 ttctttaagc agtccttccc cgagggcttc acatgggaga gagtcaccac atacgaagac
     4081 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
     4141 gtcaagatca gaggggtgaa cttcccatcc aacggccctg tgatgcagaa gaaaacactc
     4201 ggctgggagg cctccaccga gaccctgtac cccgctgacg gcggcctgga aggcagagcc
     4261 gacatggccc tgaagctcgt gggcgggggc cacctgatct gcaacttgaa gaccacatac
     4321 agatccaaga aacccgctaa gaacctcaag atgcccggcg tctactatgt ggacagaaga
     4381 ctggaaagaa tcaaggaggc cgacaaagag acctacgtcg agcagcacga ggtggctgtg
     4441 gccagatact gcgacctccc tagcaaactg gggcacagac tcgagaagtt ttctcaagaa
     4501 cagatcggcg aaaacattgt gtgcagggtc atttgtacca cgggtcaaat tcccatccga
     4561 gatttgtcag ctgatatttc acaagtgctt aaggaaaaac gatccataaa gaaagtttgg
     4621 acatttggta gaaacccagc ctgtgactat catttaggaa acatttcaag actgtcaaat
     4681 aagcatttcc aaatactact aggagaagac ggtaaccttt tattgaatga catttccact
     4741 aatgggacct ggttaaatgg gcaaaaagtc gagaagaaca gcaatcagtt actgtcccaa
     4801 ggtgatgaaa taaccgttgg tgtaggcgtg gaatcagata ttttatctct ggtcattttc
     4861 ataaacgaca aatttaagca gtgcctggag cagaacaaag ttgatcgcgg taccagtgct
     4921 ggtggtagtg ctggtggtag tgctggtggt agtgctggtg gtagtgctgg tggttccggc
     4981 agtgctggtg gtagtgctgg tggtagtacc agtgctggtg gtagtgctgg tggtagtgct
     5041 ggtggtagtg ctggtggtag tgctggtggt tccggcagtg ctggtggtag tgctggtggt
     5101 agtaccagtg ctggtggtag tgctggtggt agtgctggtg gtagtgctgg tggtagtgct
     5161 ggtggttccg gcagtgctgg tggtagtgct ggtggtagta ccagtgctgg tggtagtgct
     5221 ggtggtagtg ctggtggtag tgctggtggt agtgctggtg gttccggcag tgctggtggt
     5281 agtgctggtg gtagtaccag tgctggtggt agtgctggtg gtagtgctgg tggtagtgct
     5341 ggtggtagtg ctggtggttc cggcagtgct ggtggtagtg ctggtggtag taccagtgct
     5401 ggtggtagtg ctggtggtag tgctggtggt agtgctggtg gtagtgctgg tggttccggc
     5461 agtgctggtg gtagtgctgg tggtagtacc agtgctggtg gtagtgctgg tggtagtgct
     5521 ggtggtagtg ctggtggtag tgctggtggt tccggcagtg ctggtggtag tgctggtggt
     5581 agtaccagtg ctggtggtag tgctggtggt agtgctggtg gtagtgctgg tggtagtgct
     5641 ggtggttccg gaatggtgag tgtgattaaa ccagagatga agatgaggta ctacatggac
     5701 ggctccgtca atgggcatga gttcacaatt gaaggtgaag gcacaggcag accttacgag
     5761 ggacatcaag agatgacact acgcgtcaca atggccgagg gcgggccaat gcctttcgcg
     5821 tttgacttag tgtcacacgt gttctgttac ggccacagag tatttactaa atatccagaa
     5881 gagataccag actatttcaa acaagcattt cctgaaggcc tgtcatggga aaggtcgttg
     5941 gagttcgaag atggtgggtc cgcttcagtc agtgcgcata taagccttag aggaaacacc
     6001 ttctaccaca aatccaaatt tactggggtt aactttcctg ccgatggtcc tatcatgcaa
     6061 aaccaaagtg ttgattggga gccatcaacc gagaaaatta ctgccagcga cggagttctg
     6121 aagggtgatg ttacgatgta cctaaaactt gaaggaggcg gcaatcacaa atgccaattc
     6181 aagactactt acaaggcggc aaaagagatt cttgaaatgc caggagacca ttacatcggc
     6241 catcgcctcg tcaggaaaac cgaaggcaac attactgagc aggtagaaga tgcagtagct
     6301 cattccgcgg ccgcattgag gcgcgcgacg ctggttgact ctagactgca gctgcctcct
     6361 ctggaacgcc tgactctgga ttaagtcgag ggcggcggcc gcggctctag aggatccgtt
     6421 aactaactta agctagcaac ggtttccctc tagcgggatc aattccgccc ccccccccta
     6481 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     6541 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     6601 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     6661 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     6721 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     6781 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     6841 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     6901 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     6961 cgaggttaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     7021 acgataatac catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     7081 cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     7141 tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     7201 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     7261 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     7321 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     7381 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     7441 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     7501 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     7561 ccgtcaccgc cgacgtcgag tgcccgaagg accgcgcgac ctggtgcatg acccgcaagc
     7621 ccggtgcctg actagagtta acatcgaggg atcaagctta tcgataatca acctctggat
     7681 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     7741 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     7801 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     7861 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     7921 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     7981 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     8041 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     8101 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     8161 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     8221 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgaga cctggaaaaa
     8281 catggagcaa tcacaagtag caacacagca gctaccaatg ctgcttgtgc ctggctagaa
     8341 gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg
     8401 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     8461 ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc
     8521 tacttccctg attggcagaa ctacacacca gggccaggga tcagatatcc actgaccttt
     8581 ggatggtgct acaagctagt accagttgag caagagaagg tagaagaagc caatgaagga
     8641 gagaacaccc gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa
     8701 gtattagagt ggaggtttga cagccgccta gcatttcatc acatggcccg agagctgcat
     8761 ccggactgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     8821 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     8881 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     8941 aaaatctcta gcagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     9001 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     9061 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     9121 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     9181 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     9241 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     9301 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     9361 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt
     9421 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     9481 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     9541 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     9601 ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat
     9661 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     9721 tccccaggct ccccaggcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     9781 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     9841 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     9901 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     9961 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
    10021 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
    10081 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
    10141 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc
    10201 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt
    10261 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac
    10321 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag
    10381 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
    10441 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc
    10501 gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg
    10561 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
    10621 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
    10681 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
    10741 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
    10801 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
    10861 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
    10921 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
    10981 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
    11041 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
    11101 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
    11161 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
    11221 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
    11281 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
    11341 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
    11401 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
    11461 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
    11521 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
    11581 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
    11641 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
    11701 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
    11761 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
    11821 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
    11881 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
    11941 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
    12001 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
    12061 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
    12121 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
    12181 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
    12241 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
    12301 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
    12361 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
    12421 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
    12481 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
    12541 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
    12601 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
    12661 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
    12721 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
    12781 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
    12841 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
    12901 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
    12961 ccacctgacg tc
//
