LOCUS       sequence-245279-       10523 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd gen lentiviral expression of ChR2 with mutation fused to EYFP 
            driven by EF1a promoter for optogenetic activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10523)
  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 10523)
  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: plenti2-EF1a-ChR2-EYFP-WRPE
FEATURES             Location/Qualifiers
     source          1..10523
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    846..1079
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /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        1693..2871
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1924..2862
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2819..2839
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    2919..3036
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      3114..3123
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3120..4046
                     /codon_start=1
                     /product="N-terminal fragment of channelrhodopsin-2 from 
                     the alga Chlamydomonas reinhardtii (Nagel et al., 2003)"
                     /label=ChR2
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIHLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      4056..4065
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4062..4064,4065..4067,4068..4781)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4107..4128)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4368..4387)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4715..4736
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4799..5387
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4852..4872)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5270..5281)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    5555..5573
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5597..5615
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5639..5657
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5681..5699
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        5728..5942
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     5863..5882
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     LTR             5961..6141
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     CDS             6167..6208
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6218..6235
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6258..6275)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6264..6488
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6534..6962
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6621..6640)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6831..6852
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6971..6991)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6976..7305
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7156..7291
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        7353..7400
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7419..7817
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    7975..8096
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8012..8031)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8066..8085
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8145..8161)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8145..8161)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8158..8180)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8169..8185
                     /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(8193..8199,8200..8217,8218..8223))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8238..8259
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8376..8393)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8536..9124)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8616..8635)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9295..10086,10087..10155))
                     /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     9918..9937
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10156..10260)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        10297..10523
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     1501 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
     1561 cggtatcgat cacgagacta gcctcgacca tcgcgatgtc gacgataagc tttgcaaaga
     1621 tggataaagt tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa
     1681 ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     1741 gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta
     1801 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     1861 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     1921 caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc
     1981 gtgccttgaa ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt
     2041 ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga
     2101 gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc
     2161 tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg
     2221 ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg
     2281 gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg
     2341 cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct
     2401 gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc
     2461 cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc
     2521 tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa
     2581 agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc
     2641 aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg
     2701 ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg
     2761 cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc
     2821 aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg aggaatttcg
     2881 acatttaaat ttaattaatc tcgacggtat cggttaactt ttaaaagaaa aggggggatt
     2941 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     3001 gaattacaaa aacaaattac aaaaattcaa aattttatcg atcacgagac tagcctcgag
     3061 gtttaaacta cgggatcttc gaaggcctaa gcttacgcgc gcgtcctagc gctgccacca
     3121 tggattatgg aggcgccctg agtgccgttg ggcgcgagct gctatttgta acgaacccag
     3181 tagtcgtcaa tggctctgta cttgtgcctg aggaccagtg ttactgcgcg ggctggattg
     3241 agtcgcgtgg cacaaacggt gcccaaacgg cgtcgaacgt gctgcaatgg cttgctgctg
     3301 gcttctccat cctactgctt atgttttacg cctaccaaac atggaagtca acctgcggct
     3361 gggaggagat ctatgtgtgc gctatcgaga tggtcaaggt gattctcgag ttcttcttcg
     3421 agtttaagaa cccgtccatg ctgtatctag ccacaggcca ccgcgtccag tggttgcgtt
     3481 acgccgagtg gcttctcacc tgcccggtca ttctcattca cctgtcaaac ctgacgggct
     3541 tgtccaacga ctacagcagg cgcaccatgg gtctgcttgt gtctgatatt ggcacaattg
     3601 tgtggggcgc cacttccgcc atggccaccg gatacgtcaa ggtcatcttc ttctgcctgg
     3661 gtctgtgtta tggtgctaac acgttctttc acgctgccaa ggcctacatc gagggttacc
     3721 acaccgtgcc gaagggccgg tgtcgccagg tggtgactgg catggcttgg ctcttcttcg
     3781 tatcatgggg tatgttcccc atcctgttca tcctcggccc cgagggcttc ggcgtcctga
     3841 gcgtgtacgg ctccaccgtc ggccacacca tcattgacct gatgtcgaag aactgctggg
     3901 gtctgctcgg ccactacctg cgcgtgctga tccacgagca tatcctcatc cacggcgaca
     3961 ttcgcaagac caccaaattg aacattggtg gcactgagat tgaggtcgag acgctggtgg
     4021 aggacgaggc cgaggctggc gcggtacccg cggccgccac catggtgagc aagggcgagg
     4081 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     4141 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     4201 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct
     4261 acggcctgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     4321 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
     4381 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     4441 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
     4501 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
     4561 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     4621 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg
     4681 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     4741 ccgccgggat cactctcggc atggacgagc tgtacaagta atagactagt catatgataa
     4801 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     4861 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     4921 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     4981 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     5041 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     5101 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     5161 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     5221 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     5281 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     5341 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc ggtacgtatg
     5401 gccaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     5461 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat gggatcaatt
     5521 caccatgcta gtggatcccc ctcgagttta ccactcccta tcagtgatag agaaaagtga
     5581 aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc
     5641 cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa
     5701 aagtgaaagt cggggctgca ggaattcgaa cgctgacgtc atcaacccgc tccaaggaat
     5761 cgcgggccca gtgtcactag gcgggaacac ccagcgcgcg tgcgccctgg caggaagatg
     5821 gctgtgaggg acaggggagt ggcgccctgc aatatttgca tgtcgctatg tgttctggga
     5881 aatcaccata aacgtgaaat gtctttggat ttgggaatct tataagttct gtatgagacc
     5941 acagatctaa gcttatcgat gggtctctct ggttagacca gatctgagcc tgggagctct
     6001 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     6061 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     6121 cagtgtggaa aatctctagc agcatctaga gggcccgcgg ttcgaaggta agcctatccc
     6181 taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt
     6241 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     6301 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     6361 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     6421 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     6481 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     6541 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     6601 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     6661 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     6721 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     6781 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     6841 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     6901 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     6961 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     7021 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     7081 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     7141 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     7201 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     7261 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     7321 tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata
     7381 tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagcct ttgtctcaag
     7441 aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag
     7501 actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg
     7561 tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg
     7621 cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga
     7681 gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag
     7741 tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt
     7801 atgtgtggga gggctaagca cttcgtggcc gaggagcagg actgacacgt gctacgagat
     7861 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     7921 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     7981 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     8041 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     8101 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     8161 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     8221 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     8281 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     8341 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     8401 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     8461 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccaggaacc gtaaaaaggc
     8521 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     8581 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     8641 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     8701 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     8761 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     8821 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     8881 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     8941 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     9001 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     9061 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     9121 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     9181 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     9241 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     9301 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     9361 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     9421 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     9481 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     9541 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     9601 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     9661 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     9721 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     9781 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     9841 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     9901 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     9961 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
    10021 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
    10081 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
    10141 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
    10201 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
    10261 cacatttccc cgaaaagtgc cacctgacgt ccttaatgta gtcttatgca atactcttgt
    10321 agtcttgcaa catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc
    10381 gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga
    10441 cgggtctgac atggattgga cgaaccactg aattgccgca ttgcagagat attgtattta
    10501 agtgcctagc tcgatacata aac
//
