LOCUS       sequence-239640-       11571 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 11571)
  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 11571)
  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..11571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2682..3859
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2913..3850
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3807..3827
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3866..3875
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3872..4798
                     /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             4811..6238
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYE
                     GTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMN
                     FEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVL
                     KGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYER
                     SEGRHHLFLYGMDELYK"
     misc_feature    6263..6851
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6316..6336)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6734..6745)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6854..6870)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6855..6871)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7376..7556
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7582..7599)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7588..7812
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7858..8286
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7945..7964)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8155..8176
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8295..8315)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8300..8629
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8480..8615
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8542..8561
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8677..8724
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8743..9117
                     /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    9247..9368
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9284..9303)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9338..9357
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9417..9433)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9417..9433)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9430..9452)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9441..9457
                     /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(9465..9471,9472..9489,9490..9495))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9510..9531
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9648..9665)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9819..10407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9899..9918)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10578..11369,11370..11438))
                     /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     11201..11220
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11439..11543)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta atgcaaagat ggataaagtt ttaaacagag aggaatcttt
     2641 gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt
     2701 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     2761 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     2821 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     2881 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
     2941 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccact ggctgcagta
     3001 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc
     3061 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg
     3121 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat
     3181 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc
     3241 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg
     3301 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
     3361 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
     3421 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
     3481 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg
     3541 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg
     3601 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag
     3661 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
     3721 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct
     3781 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
     3841 tccatttcag gtgtcgtgac gtacggccac catggactat ggcggcgctt tgtctgccgt
     3901 cggacgcgaa cttttgttcg ttactaatcc tgtggtggtg aacgggtccg tcctggtccc
     3961 tgaggatcaa tgttactgtg ccggatggat tgaatctcgc ggcacgaacg gcgctcagac
     4021 cgcgtcaaat gtcctgcagt ggcttgcagc aggattcagc attttgctgc tgatgttcta
     4081 tgcctaccaa acctggaaat ctacatgcgg ctgggaggag atctatgtgt gcgccattga
     4141 aatggttaag gtgattctcg agttcttttt tgagtttaag aatccctcta tgctctacct
     4201 tgccacagga caccgggtgc agtggctgcg ctatgcagag tggctgctca cttgtcctgt
     4261 catccttatc cgcctgagca acctcaccgg cctgagcaac gactacagca ggagaaccat
     4321 gggactcctt gtctcagaca tcgggactat cgtgtggggg gctaccagcg ccatggcaac
     4381 cggctatgtt aaagtcatct tcttttgtct tggattgtgc tatggcgcga acacattttt
     4441 tcacgccgcc aaagcatata tcgagggtta tcatactgtg ccaaagggtc ggtgccgcca
     4501 ggtcgtgacc ggcatggcat ggctgttttt cgtgagctgg ggtatgttcc caattctctt
     4561 cattttgggg cccgaaggtt ttggcgtcct gagcgtctat ggctccaccg taggtcacac
     4621 gattattgat ctgatgagta aaaattgttg ggggttgttg ggacactacc tgcgcgtcct
     4681 gatccacgag cacatattga ttcacggaga tatccgcaaa accaccaaac tgaacatcgg
     4741 cggaacggag atcgaggtcg agactctcgt cgaagacgaa gccgaggccg gagccgtgcc
     4801 agccgcggcc gtgagcaagg gcgaggaggt catcaaagag ttcatgcgct tcaaggtgcg
     4861 catggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     4921 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc
     4981 ctgggacatc ctgtcccccc agttcatgta cggctccaag gcgtacgtga agcaccccgc
     5041 cgacatcccc gattacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     5101 gaacttcgag gacggcggtc tggtgaccgt gacccaggac tcctccctgc aggacggcac
     5161 gctgatctac aaggtgaaga tgcgcggcac caacttcccc cccgacggcc ccgtaatgca
     5221 gaagaagacc atgggctggg aggcctccac cgagcgcctg tacccccgcg acggcgtgct
     5281 gaagggcgag atccaccagg ccctgaagct gaaggacggc ggccactacc tggtggagtt
     5341 caagaccatc tacatggcca agaagcccgt gcaactgccc ggctactact acgtggacac
     5401 caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgagcgctc
     5461 cgagggccgc caccacctgt tcctggggca tggcaccggc agcaccggca gcggcagctc
     5521 cggcaccgcc tcctccgagg acaacaacat ggccgtcatc aaagagttca tgcgcttcaa
     5581 ggtgcgcatg gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     5641 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc
     5701 cttcgcctgg gacatcctgt ccccccagtt catgtacggc tccaaggcgt acgtgaagca
     5761 ccccgccgac atccccgatt acaagaagct gtccttcccc gagggcttca agtgggagcg
     5821 cgtgatgaac ttcgaggacg gcggtctggt gaccgtgacc caggactcct ccctgcagga
     5881 cggcacgctg atctacaagg tgaagatgcg cggcaccaac ttcccccccg acggccccgt
     5941 aatgcagaag aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg
     6001 cgtgctgaag ggcgagatcc accaggccct gaagctgaag gacggcggcc actacctggt
     6061 ggagttcaag accatctaca tggccaagaa gcccgtgcaa ctgcccggct actactacgt
     6121 ggacaccaag ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     6181 gcgctccgag ggccgccacc acctgttcct gtacggcatg gacgagctgt acaagtaaga
     6241 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     6301 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     6361 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     6421 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     6481 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     6541 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     6601 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     6661 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     6721 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     6781 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     6841 cgcctccccg catcgatacc gtcgacctcg agacctagaa aaacatggag caatcacaag
     6901 tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga
     6961 ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt
     7021 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     7081 aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattggca
     7141 gaactacaca ccagggccag ggatcagata tccactgacc tttggatggt gctacaagct
     7201 agtaccagtt gagcaagaga aggtagaaga agccaatgaa ggagagaaca cccgcttgtt
     7261 acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtattag agtggaggtt
     7321 tgacagccgc ctagcatttc atcacatggc ccgagagctg catccggact gtactgggtc
     7381 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     7441 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     7501 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagggc
     7561 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     7621 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     7681 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     7741 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     7801 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     7861 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     7921 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     7981 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     8041 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     8101 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     8161 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     8221 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     8281 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     8341 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     8401 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     8461 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     8521 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     8581 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     8641 agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag
     8701 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt
     8761 gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg
     8821 ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg
     8881 accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg
     8941 tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc
     9001 cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc
     9061 gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac
     9121 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     9181 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     9241 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     9301 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     9361 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     9421 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     9481 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     9541 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     9601 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     9661 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     9721 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     9781 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     9841 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     9901 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     9961 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
    10021 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
    10081 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
    10141 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
    10201 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
    10261 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
    10321 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
    10381 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
    10441 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
    10501 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
    10561 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
    10621 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
    10681 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
    10741 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
    10801 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
    10861 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
    10921 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
    10981 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
    11041 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
    11101 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
    11161 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
    11221 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
    11281 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
    11341 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
    11401 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
    11461 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
    11521 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga c
//
