LOCUS       sequence-239642-       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        96..1274
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          327..1265
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1222..1242
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1294..1318
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1322..2248
                     /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             2261..2968
                     /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(2403..2421)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2671..2690
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(2970..2994)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3002..3043
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3094..3682
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3147..3167)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3565..3576)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3754..3987
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4065..4186
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4102..4121)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4156..4175
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4226..4361
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4381..4400)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4382..4400)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4410..4427)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4410..4426)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4419..4441)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4568..5023
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4655..4674)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4865..4886
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5049..5153
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5154..5222,5223..6014)
                     /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(5372..5391)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6185..6773
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6674..6693
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6927..6944
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7061..7082
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7097..7102,7103..7120,7121..7127)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7135..7151
                     /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     7140..7162
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7159..7175
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7159..7175
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7194..7214
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7196..7214
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7242..7468
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7469..7649
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7696..7821
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8314..8547
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8732..8776
                     /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"
     promoter        9007..9217
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        9029..9225
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      9076..9211
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             9257..9859
                     /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(9257..9276)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9753..9773
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9902..10019
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ctcgaggcct gcaggtgcaa agatggataa agttttaaac agagaggaat ctttgcagct
       61 aatggacctt ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt cagtgggcag
      121 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg
      181 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
      241 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
      301 gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc
      361 tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga
      421 ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag
      481 gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc
      541 gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa
      601 atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc
      661 aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt
      721 cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg
      781 ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc
      841 cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc
      901 ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg
      961 gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact
     1021 ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt
     1081 cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg
     1141 agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg
     1201 agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat
     1261 ttcaggtgtc gtgaggctag catcgattga tcaacaagtt tgtacaaaaa agcaggctac
     1321 catggactat ggcggcgctt tgtctgccgt cggacgcgaa cttttgttcg ttactaatcc
     1381 tgtggtggtg aacgggtccg tcctggtccc tgaggatcaa tgttactgtg ccggatggat
     1441 tgaatctcgc ggcacgaacg gcgctcagac cgcgtcaaat gtcctgcagt ggcttgcagc
     1501 aggattcagc attttgctgc tgatgttcta tgcctaccaa acctggaaat ctacatgcgg
     1561 ctgggaggag atctatgtgt gcgccattga aatggttaag gtgattctcg agttcttttt
     1621 tgagtttaag aatccctcta tgctctacct tgccacagga caccgggtgc agtggctgcg
     1681 ctatgcagag tggctgctca cttgtcctgt catccttatc cgcctgagca acctcaccgg
     1741 cctgagcaac gactacagca ggagaaccat gggactcctt gtctcagaca tcgggactat
     1801 cgtgtggggg gctaccagcg ccatggcaac cggctatgtt aaagtcatct tcttttgtct
     1861 tggattgtgc tatggcgcga acacattttt tcacgccgcc aaagcatata tcgagggtta
     1921 tcatactgtg ccaaagggtc ggtgccgcca ggtcgtgacc ggcatggcat ggctgttttt
     1981 cgtgagctgg ggtatgttcc caattctctt cattttgggg cccgaaggtt ttggcgtcct
     2041 gagcgtctat ggctccaccg taggtcacac gattattgat ctgatgagta aaaattgttg
     2101 ggggttgttg ggacactacc tgcgcgtcct gatccacgag cacatattga ttcacggaga
     2161 tatccgcaaa accaccaaac tgaacatcgg cggaacggag atcgaggtcg agactctcgt
     2221 cgaagacgaa gccgaggccg gagccgtgcc agcggccgcc gtgagcaagg gcgaggagga
     2281 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
     2341 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     2401 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
     2461 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
     2521 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     2581 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     2641 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     2701 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
     2761 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
     2821 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
     2881 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     2941 cggcggcatg gacgagctgt acaagtaaca cccagctttc ttgtacaaag tggttgatat
     3001 cggtaagcct atccctaacc ctctcctcgg tctcgattct acgtagtaat gaactagtac
     3061 cggttaagtc gacaatcaac gcgttaagtc gacaatcaac ctctggatta caaaatttgt
     3121 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3181 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3241 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     3301 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     3361 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     3421 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     3481 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     3541 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     3601 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     3661 tccctttggg ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag
     3721 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     3781 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     3841 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     3901 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     3961 tttagtcagt gtggaaaatc tctagcagta cgtatagtag ttcatgtcat cttattattc
     4021 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     4081 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4141 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     4201 agctatcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4261 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     4321 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg
     4381 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     4441 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4501 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     4561 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4621 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4681 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4741 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4801 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4861 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4921 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4981 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     5041 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5101 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5161 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5221 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5281 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5341 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5401 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5461 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5521 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5581 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5641 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5701 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5761 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5821 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5881 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5941 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6001 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6061 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6121 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6181 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6241 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6301 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6361 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6421 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6481 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6541 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6601 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6661 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6721 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6781 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6841 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     6901 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     6961 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     7021 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     7081 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     7141 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     7201 accctcacta aagggaacaa aagctggagc tgcaagctta atgtagtctt atgcaatact
     7261 cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag gagagaaaaa
     7321 gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat taggaaggca
     7381 acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca gagatattgt
     7441 atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga tctgagcctg
     7501 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     7561 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     7621 cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa
     7681 gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga
     7741 ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag
     7801 agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa
     7861 attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc
     7921 agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga
     7981 caaatactgg gacagctaca accatccctt cagacaggat cagaagaact tagatcatta
     8041 tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag
     8101 gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg
     8161 gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata
     8221 taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag
     8281 agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg
     8341 agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt
     8401 attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca
     8461 tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga
     8521 aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg
     8581 caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa
     8641 tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct taatacactc
     8701 cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga
     8761 taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt
     8821 attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat
     8881 agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc
     8941 gaggggaccc atgcattgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     9001 tccccagcag gcagaagtat gcaaagcatg cgtctcaatt agtcagcaac catagtcccg
     9061 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     9121 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     9181 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct ttctagaggt
     9241 accaccggag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     9301 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     9361 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     9421 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg ccgtggcggt
     9481 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     9541 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     9601 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     9661 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     9721 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     9781 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     9841 caagcccggt gccagttaac gaattctaga tcttgagaca aatggcagta ttcatccaca
     9901 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     9961 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc
    10021 gggtttatta cagggacagc agagatccac tttggcgccg g
//
