LOCUS       sequence-203589-       10521 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac plasmid encoding channelrhodopsin 2 fused to EYFP in an 
            inverse orientation flanked by loxP and lox2722 sites under the 
            control of the EF1a promotor..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10521)
  AUTHORS   Klapper SD, Sauter EJ, Swiersy A, Hyman MAE, Bamann C, Bamberg E, 
            Busskamp V
  TITLE     On-demand optogenetic activation of human stem-cell-derived neurons.
  JOURNAL   Sci Rep. 2017 Oct 31;7(1):14450. doi: 10.1038/s41598-017-14827-6.
  PUBMED    29089561
REFERENCE   2  (bases 1 to 10521)
  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 Rep."; 
            date: "2017-10-31"; volume: "7(1):14450. doi"; pages: " 
            10.1038/s41598-017-14827-6"
COMMENT     Sequence Label: PB-eDIO-ChR2-EYFP
FEATURES             Location/Qualifiers
     source          1..10521
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(115..570)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(252..273)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     464..483
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     697..719
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     711..728
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     712..728
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   858..920
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1421..2599
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1652..2590
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2547..2567
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(2630..2663)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(2714..2747)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(2759..3472,3473..3475,3476..3478))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2804..2825)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3153..3172
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3412..3433
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3475..3484
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3494..4420)
                     /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"
     regulatory      4417..4426
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    4431..4464
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    4515..4548
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4562..5150
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4615..4635)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5033..5044)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5157..5381
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        5393..6604
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6441..6460
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      6634..6643
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6640..7038
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYAWEG"
     polyA_signal    7077..7198
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7114..7133)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7168..7187
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7959..7993
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8484..8500)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8484..8500)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8497..8519)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8508..8524
                     /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(8532..8538,8539..8556,8557..8562))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8577..8598
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8715..8732)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8886..9474)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8966..8985)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9645..10436,10437..10505))
                     /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     10268..10287
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(10506..10521,1..89))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
       61 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
      121 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
      181 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
      241 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
      301 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
      361 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
      421 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
      481 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
      541 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
      601 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
      661 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
      721 cggccagtga gcgcgcctcg ttcattcacg tttttgaacc cgtggaggac gggcagactc
      781 gcggtgcaaa tgtgttttac agcgtgatgg agcagatgaa gatgctcgac acgctgcaga
      841 acacgcagct agattaaccc tagaaagata atcatattgt gacgtacgtt aaagataatc
      901 atgtgtaaaa ttgacgcatg tgttttatcg gtctgtatat cgaggtttat ttattaattt
      961 gaatagatat taagttttat tatatttaca cttacatact aataataaat tcaacaaaca
     1021 atttatttat gtttatttat ttattaaaaa aaacaaaaac tcaaaatttc ttctataaag
     1081 taacaaaact tttatgaggg acagcccccc cccaaagccc ccagggatgt aattacgtcc
     1141 ctcccccgct agggggcagc agcgagccgc ccggggctcc gctccggtcc ggcgctcccc
     1201 ccgcatcccc gagccggcag cgtgcgggga cagcccgggc acggggaagg tggcacggga
     1261 tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg cagacacctg gggggatacg
     1321 gggaaaaggc ctgtaagctt tgcaaagatg gataaagttt taaacagaga ggaatctttg
     1381 cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg
     1441 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
     1501 cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
     1561 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
     1621 ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc
     1681 tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct ggctgcagta
     1741 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc
     1801 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg
     1861 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat
     1921 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc
     1981 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg
     2041 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
     2101 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
     2161 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
     2221 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg
     2281 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg
     2341 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag
     2401 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
     2461 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct
     2521 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
     2581 tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa taacttcgta
     2641 taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc tattagcaat
     2701 atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc gcgccctatt
     2761 acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     2821 ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc ggactggtag ctcaggtagt
     2881 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     2941 cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt
     3001 tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt
     3061 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     3121 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     3181 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     3241 tcatgtggtc ggggtagcgg gcgaagcact gcaggccgta gccgaaggtg gtcacgaggg
     3301 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt
     3361 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     3421 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg
     3481 tggcggccgc tggcacggct ccggcctcgg cttcgtcttc gacgagagtc tcgacctcga
     3541 tctccgttcc gccgatgttc agtttggtgg ttttgcggat atctccgtga atcaatatgt
     3601 gctcgtggat caggacgcgc aggtagtgtc ccaacaaccc ccaacaattt ttactcatca
     3661 gatcaataat cgtgtgacct acggtggagc catagacgct caggacgcca aaaccttcgg
     3721 gccccaaaat gaagagaatt gggaacatac cccagctcac gaaaaacagc catgccatgc
     3781 cggtcacgac ctggcggcac cgaccctttg gcacagtatg ataaccctcg atatatgctt
     3841 tggcggcgtg aaaaaatgtg ttcgcgccat agcacaatcc aagacaaaag aagatgactt
     3901 taacatagcc ggttgccatg gcgctggtag ccccccacac gatagtcccg atgtctgaga
     3961 caaggagtcc catggttctc ctgctgtagt cgttgctcag gccggtgagg ttgctcaggc
     4021 ggataaggat gacaggacaa gtgagcagcc actctgcata gcgcagccac tgcacccggt
     4081 gtcctgtggc aaggtagagc atagagggat tcttaaactc aaaaaagaac tcgagaatca
     4141 ccttaaccat ttcaatggcg cacacataga tctcctccca gccgcatgta gatttccagg
     4201 tttggtaggc atagaacatc agcagcaaaa tgctgaatcc tgctgcaagc cactgcagga
     4261 catttgacgc ggtctgagcg ccgttcgtgc cgcgagattc aatccatccg gcacagtaac
     4321 attgatcctc agggaccagg acggacccgt tcaccaccac aggattagta acgaacaaaa
     4381 gttcgcgtcc gacggcagac aaagcgccgc catagtccat ggtggctagc ataacttcgt
     4441 ataaagtatc ctatacgaag ttatttgcct taacccagaa attatcactg ttattcttta
     4501 gaatggtgca aagaataact tcgtataatg tatgctatac gaagttatga attcggatcc
     4561 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     4621 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     4681 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     4741 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     4801 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     4861 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     4921 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     4981 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     5041 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     5101 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc gaattcctgt
     5161 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     5221 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     5281 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     5341 agacaatagc aggcatgctg gggatgcggt gggctctatg gatcgatgat ctggcctccg
     5401 cgccgggttt tggcgcctcc cgcgggcgcc cccctcctca cggcgagcgc tgccacgtca
     5461 gacgaagggc gcagcgagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc
     5521 tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac
     5581 ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag
     5641 tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat
     5701 aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc
     5761 ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg
     5821 ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct
     5881 gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa
     5941 tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg
     6001 aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgaggcc ttcgctaatg
     6061 cgggaaagct cttattcggg tgagatgggc tggggcacca tctggggacc ctgacgtgaa
     6121 gtttgtcact gactggagaa ctcggtttgt cgtctgttgc gggggcggca gttatggcgg
     6181 tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc
     6241 acccgttctg ttggcttata atgcagggtg gggccacctg ccggtaggtg tgcggtaggc
     6301 ttttctccgt cgcaggacgc agggttcggg cctagggtag gctctcctga atcgacaggc
     6361 gccggacctc tggtgagggg agggataagt gaggcgtcag tttctttggt cggttttatg
     6421 tacctatctt cttaagtagc tgaagctccg gttttgaact atgcgctcgg ggttggcgag
     6481 tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa
     6541 ttttcagtgt tagactagta aattgtccgc taaattctgg ccgtttttgg cttttttgtt
     6601 agacgaagct tgggctgcag gtcgactcta gatgccacca tggccaagcc tttgtctcaa
     6661 gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa
     6721 gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat
     6781 gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct
     6841 gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg
     6901 agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata
     6961 gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt
     7021 tatgcgtggg agggctaagt cgacaatcaa cctctggatt acaaaatttg tgggttaact
     7081 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     7141 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     7201 atgtctggaa ttgactcaaa tgatgtcaat tagtctatca gaagctatct ggtctccctt
     7261 ccgggggaca agacatccct gtttaatatt taaacagcag tgttcccaaa ctgggttctt
     7321 atatcccttg ctctggtcaa ccaggttgca gggtttcctg tcctcacagg aacgaagtcc
     7381 ctaaagaaac agtggcagcc aggtttagcc ccggaattga ctggattcct tttttagggc
     7441 ccattggtat ggctttttcc ccgtatcccc ccaggtgtct gcaggctcaa agagcagcga
     7501 gaagcgttca gaggaaagcg atcccgtgcc accttccccg tgcccgggct gtccccgcac
     7561 gctgccggct cggggatgcg gggggagcgc cggaccggag cggagccccg ggcggctcgc
     7621 tgctgccccc tagcggggga gggacgtaat tacatccctg ggggctttgg ggggggctgt
     7681 ccctgatatc tataacaaga aaatatatat ataataagtt atcacgtaag tagaacatga
     7741 aataacaata taattatcgt atgagttaaa tcttaaaagt cacgtaaaag ataatcatgc
     7801 gtcattttga ctcacgcggt cgttatagtt caaaatcagt gacacttacc gcattgacaa
     7861 gcacgcctca cgggagctcc aagcggcgac tgagatgtcc taaatgcaca gcgacggatt
     7921 cgcgctattt agaaagagag agcaatattt caagaatgca tgcgtcaatt ttacgcagac
     7981 tatctttcta gggttaatct agctgcatca ggatcatatc gtcgggtctt ttttccggct
     8041 cagtcatcgc ccaagctggc gctatctggg catcggggag gaagaagccc gtgccttttc
     8101 ccgcgaggtt gaagcggcat ggaaagagtt tgccgaggat gactgctgct gcattgacgt
     8161 tgagcgaaaa cgcacgttta ccatgatgat tcgggaaggt gtggccatgc acgcctttaa
     8221 cggtgaactg ttcgttcagg ccacctggga taccagttcg tcgcggcttt tccggacaca
     8281 gttccggatg gtcagcccga agcgcatcag caacccgaac aataccggcg acagccggaa
     8341 ctgccgtgcc ggtgtgcaga ttaatgacag cggtgcggcg ctgggatatt acgtcagcga
     8401 ggacgggtat cctggctgga tgccgcagaa atggacatgg ataccccgtg agttacccgg
     8461 cgggcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     8521 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     8581 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     8641 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     8701 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     8761 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     8821 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     8881 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     8941 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     9001 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     9061 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     9121 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     9181 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     9241 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     9301 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     9361 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     9421 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     9481 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     9541 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     9601 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     9661 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     9721 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     9781 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     9841 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     9901 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     9961 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
    10021 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
    10081 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
    10141 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
    10201 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
    10261 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
    10321 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
    10381 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
    10441 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
    10501 ctcatactct tcctttttca a
//
