LOCUS       sequence-245091-       10061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd gen lentiviral expression of humanized ChR2 with H134R mutation 
            fused to mCherry driven by EF1a promoter for optogenetic activation 
            with puro selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10061)
  AUTHORS   Osaki T, Uzel SGM, Kamm RD
  TITLE     Microphysiological 3D model of amyotrophic lateral sclerosis (ALS) 
            from human iPS-derived muscle cells and optogenetic motor neurons.
  JOURNAL   Sci Adv. 2018 Oct 10;4(10):eaat5847. doi: 10.1126/sciadv.aat5847. 
            eCollection 2018 Oct.
  PUBMED    30324134
REFERENCE   2  (bases 1 to 10061)
  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: "Sci Adv."; 
            date: "2018-10-10"; volume: "4(10):eaat5847. doi"; pages: " 
            10.1126/sciadv.aat5847. eCollection 2018 Oct"
COMMENT     Sequence Label: pLEX307-EF1a-ChR2[H134R]-mCherry-Puro-WPRE
FEATURES             Location/Qualifiers
     source          1..10061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        167..377
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        189..385
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      236..371
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             417..1019
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAS"
     primer_bind     complement(417..436)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     913..933
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    1062..1179
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1317..2495
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1548..2486
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2443..2463
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    2515..2539
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2543..3469
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             3482..4189
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(3624..3642)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3892..3911
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(4191..4215)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4223..4264
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    4315..4903
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4368..4388)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4786..4797)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4975..5208
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5286..5407
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5323..5342)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5377..5396
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5447..5582
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5602..5621)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5603..5621)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5631..5648)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5631..5647)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5640..5662)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5789..6244
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5876..5895)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6086..6107
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6270..6374
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6375..6443,6444..7235)
                     /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     complement(6593..6612)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7406..7994
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7895..7914
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8148..8165
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8282..8303
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8318..8323,8324..8341,8342..8348)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8356..8372
                     /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)."
     primer_bind     8361..8383
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8380..8396
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8380..8396
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8415..8435
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8417..8435
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8463..8689
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8690..8870
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8917..9042
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9535..9768
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9953..9997
                     /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 gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg
       61 gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc atgcattgca
      121 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
      181 gcaaagcatg cgtctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
      241 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
      301 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
      361 ttttggaggc ctaggctttt gcaaaaagct ttctagaggt accaccggag cttaccatga
      421 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
      481 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
      541 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
      601 gcaaggtgtg ggtcgcggac gacggcgccg ccgtggcggt ctggaccacg ccggagagcg
      661 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
      721 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
      781 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
      841 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
      901 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
      961 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gccagttaac
     1021 gaattctaga tcttgagaca aatggcagta ttcatccaca attttaaaag aaaagggggg
     1081 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
     1141 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
     1201 agagatccac tttggcgccg gctcgaggcc tgcaggtgca aagatggata aagttttaaa
     1261 cagagaggaa tctttgcagc taatggacct tctaggtctt gaaaggagtg ggaattggct
     1321 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     1381 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     1441 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     1501 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
     1561 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
     1621 ccacctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga
     1681 gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg
     1741 ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg
     1801 ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag
     1861 atagtcttgt aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg
     1921 gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc
     1981 ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc
     2041 tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg
     2101 cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc
     2161 ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct
     2221 cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt
     2281 tctcgagctt ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt
     2341 ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct
     2401 ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg
     2461 ttcaaagttt ttttcttcca tttcaggtgt cgtgaggcta gcatcgattg atcaacaagt
     2521 ttgtacaaaa aagcaggcta ccatggacta tggcggcgct ttgtctgccg tcggacgcga
     2581 acttttgttc gttactaatc ctgtggtggt gaacgggtcc gtcctggtcc ctgaggatca
     2641 atgttactgt gccggatgga ttgaatctcg cggcacgaac ggcgctcaga ccgcgtcaaa
     2701 tgtcctgcag tggcttgcag caggattcag cattttgctg ctgatgttct atgcctacca
     2761 aacctggaaa tctacatgcg gctgggagga gatctatgtg tgcgccattg aaatggttaa
     2821 ggtgattctc gagttctttt ttgagtttaa gaatccctct atgctctacc ttgccacagg
     2881 acaccgggtg cagtggctgc gctatgcaga gtggctgctc acttgtcctg tcatccttat
     2941 ccgcctgagc aacctcaccg gcctgagcaa cgactacagc aggagaacca tgggactcct
     3001 tgtctcagac atcgggacta tcgtgtgggg ggctaccagc gccatggcaa ccggctatgt
     3061 taaagtcatc ttcttttgtc ttggattgtg ctatggcgcg aacacatttt ttcacgccgc
     3121 caaagcatat atcgagggtt atcatactgt gccaaagggt cggtgccgcc aggtcgtgac
     3181 cggcatggca tggctgtttt tcgtgagctg gggtatgttc ccaattctct tcattttggg
     3241 gcccgaaggt tttggcgtcc tgagcgtcta tggctccacc gtaggtcaca cgattattga
     3301 tctgatgagt aaaaattgtt gggggttgtt gggacactac ctgcgcgtcc tgatccacga
     3361 gcacatattg attcacggag atatccgcaa aaccaccaaa ctgaacatcg gcggaacgga
     3421 gatcgaggtc gagactctcg tcgaagacga agccgaggcc ggagccgtgc cagcggccgc
     3481 cgtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     3541 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     3601 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     3661 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     3721 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     3781 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     3841 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     3901 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     3961 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     4021 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     4081 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     4141 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaac acccagcttt
     4201 cttgtacaaa gtggttgata tcggtaagcc tatccctaac cctctcctcg gtctcgattc
     4261 tacgtagtaa tgaactagta ccggttaagt cgacaatcaa cgcgttaagt cgacaatcaa
     4321 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4381 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4441 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     4501 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4561 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4621 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4681 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     4741 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     4801 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     4861 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa
     4921 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     4981 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     5041 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     5101 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     5161 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt acgtatagta
     5221 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     5281 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5341 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5401 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac
     5461 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     5521 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     5581 cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc
     5641 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     5701 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     5761 cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     5821 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     5881 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5941 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     6001 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     6061 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     6121 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     6181 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     6241 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     6301 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     6361 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     6421 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     6481 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     6541 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     6601 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     6661 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     6721 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6781 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6841 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6901 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     6961 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     7021 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     7081 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     7141 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     7201 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     7261 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     7321 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     7381 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     7441 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     7501 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     7561 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7621 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7681 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7741 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7801 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7861 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7921 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7981 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     8041 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     8101 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     8161 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     8221 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     8281 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     8341 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     8401 attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt
     8461 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
     8521 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
     8581 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
     8641 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
     8701 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     8761 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     8821 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
     8881 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
     8941 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
     9001 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
     9061 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
     9121 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
     9181 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
     9241 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
     9301 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
     9361 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
     9421 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     9481 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     9541 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     9601 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     9661 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     9721 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     9781 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     9841 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     9901 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     9961 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
    10021 ttggctgtgg tatataaaat tattcataat gatagtagga g
//
