LOCUS       sequence-197955-       10522 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 10522)
  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 10522)
  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..10522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(268..289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     480..499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     713..735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     727..744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   874..936
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1438..2615
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1669..2606
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2563..2583
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(2646..2679)
                     /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(2730..2763)
                     /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(2775..3488,3489..3491,3492..3494))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2820..2841)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3169..3188
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3428..3449
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3491..3500
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3510..4436)
                     /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      4433..4442
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    4447..4480
                     /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    4531..4564
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4578..5166
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4631..4651)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5049..5060)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5173..5397
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        5409..6620
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6457..6476
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      6650..6659
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6656..7054
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYAWEG"
     polyA_signal    7093..7214
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7130..7149)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7184..7203
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7976..8010
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8501..8517)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8501..8517)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8514..8536)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8525..8541
                     /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(8549..8555,8556..8573,8574..8579))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8594..8615
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8732..8749)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8903..9491)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8983..9002)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9662..10453,10454..10522))
                     /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     10285..10304
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgc gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaaa caaaaactca
     1081 aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc aaagccccca
     1141 gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct
     1201 ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg
     1261 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
     1321 acacctgggg ggatacgggg aaaaggcctg taagctttgc aaagatggat aaagttttaa
     1381 acagagagga atctttgcag ctaatggacc ttctaggtct tgaaaggagt gggaattggc
     1441 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
     1501 ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
     1561 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     1621 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg
     1681 tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact
     1741 tccactggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga
     1801 gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg
     1861 ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg
     1921 ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag
     1981 atagtcttgt aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg
     2041 gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc
     2101 ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc
     2161 tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg
     2221 cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc
     2281 ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct
     2341 cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt
     2401 tctcgagctt ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt
     2461 ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct
     2521 ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg
     2581 ttcaaagttt ttttcttcca tttcaggtgt cgtgaggtac cggatcctct agagtcgact
     2641 ccggaataac ttcgtatagg atactttata cgaagttatg cagaatggta gctggattgt
     2701 agctgctatt agcaatatga aacctcttaa taacttcgta tagcatacat tatacgaagt
     2761 tatggcgcgc cctattactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc
     2821 acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac
     2881 tggtagctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc
     2941 tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag
     3001 ttcaccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag
     3061 ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc
     3121 tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg
     3181 ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg
     3241 aagaagtcgt gctgcttcat gtggtcgggg tagcgggcga agcactgcag gccgtagccg
     3301 aaggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc
     3361 agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg
     3421 ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg
     3481 cccttgctca ccatggtggc ggccgctggc acggctccgg cctcggcttc gtcttcgacg
     3541 agagtctcga cctcgatctc cgttccgccg atgttcagtt tggtggtttt gcggatatct
     3601 ccgtgaatca atatgtgctc gtggatcagg acgcgcaggt agtgtcccaa caacccccaa
     3661 caatttttac tcatcagatc aataatcgtg tgacctacgg tggagccata gacgctcagg
     3721 acgccaaaac cttcgggccc caaaatgaag agaattggga acatacccca gctcacgaaa
     3781 aacagccatg ccatgccggt cacgacctgg cggcaccgac cctttggcac agtatgataa
     3841 ccctcgatat atgctttggc ggcgtgaaaa aatgtgttcg cgccatagca caatccaaga
     3901 caaaagaaga tgactttaac atagccggtt gccatggcgc tggtagcccc ccacacgata
     3961 gtcccgatgt ctgagacaag gagtcccatg gttctcctgc tgtagtcgtt gctcaggccg
     4021 gtgaggttgc tcaggcggat aaggatgaca ggacaagtga gcagccactc tgcatagcgc
     4081 agccactgca cccggtgtcc tgtggcaagg tagagcatag agggattctt aaactcaaaa
     4141 aagaactcga gaatcacctt aaccatttca atggcgcaca catagatctc ctcccagccg
     4201 catgtagatt tccaggtttg gtaggcatag aacatcagca gcaaaatgct gaatcctgct
     4261 gcaagccact gcaggacatt tgacgcggtc tgagcgccgt tcgtgccgcg agattcaatc
     4321 catccggcac agtaacattg atcctcaggg accaggacgg acccgttcac caccacagga
     4381 ttagtaacga acaaaagttc gcgtccgacg gcagacaaag cgccgccata gtccatggtg
     4441 gctagcataa cttcgtataa agtatcctat acgaagttat ttgccttaac ccagaaatta
     4501 tcactgttat tctttagaat ggtgcaaaga ataacttcgt ataatgtatg ctatacgaag
     4561 ttatgaattc ggatcctaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     4621 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     4681 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     4741 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     4801 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     4861 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     4921 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     4981 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     5041 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     5101 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     5161 ccccgcgaat tcctgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     5221 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     5281 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     5341 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggatc
     5401 gatgatctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcctcacggc
     5461 gagcgctgcc acgtcagacg aagggcgcag cgagcgtcct gatccttccg cccggacgct
     5521 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga
     5581 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac
     5641 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga
     5701 acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg
     5761 atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct
     5821 gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga
     5881 gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg
     5941 ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg
     6001 ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt
     6061 gaggccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg gcaccatctg
     6121 gggaccctga cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg
     6181 gcggcagtta tggcggtgcc gttgggcagt gcacccgtac ctttgggagc gcgcgccctc
     6241 gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc agggtggggc cacctgccgg
     6301 taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc
     6361 tcctgaatcg acaggcgccg gacctctggt gaggggaggg ataagtgagg cgtcagtttc
     6421 tttggtcggt tttatgtacc tatcttctta agtagctgaa gctccggttt tgaactatgc
     6481 gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa atgtaatcat
     6541 ttgggtcaat atgtaatttt cagtgttaga ctagtaaatt gtccgctaaa ttctggccgt
     6601 ttttggcttt tttgttagac gaagcttggg ctgcaggtcg actctagatg ccaccatggc
     6661 caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag
     6721 catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat
     6781 cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct
     6841 gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga
     6901 gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc
     6961 tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg ggattcgtga
     7021 attgctgccc tctggttatg cgtgggaggg ctaagtcgac aatcaacctc tggattacaa
     7081 aatttgtggg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     7141 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     7201 atcaatgtat cttatcatgt ctggaattga ctcaaatgat gtcaattagt ctatcagaag
     7261 ctatctggtc tcccttccgg gggacaagac atccctgttt aatatttaaa cagcagtgtt
     7321 cccaaactgg gttcttatat cccttgctct ggtcaaccag gttgcagggt ttcctgtcct
     7381 cacaggaacg aagtccctaa agaaacagtg gcagccaggt ttagccccgg aattgactgg
     7441 attccttttt tagggcccat tggtatggct ttttccccgt atccccccag gtgtctgcag
     7501 gctcaaagag cagcgagaag cgttcagagg aaagcgatcc cgtgccacct tccccgtgcc
     7561 cgggctgtcc ccgcacgctg ccggctcggg gatgcggggg gagcgccgga ccggagcgga
     7621 gccccgggcg gctcgctgct gccccctagc gggggaggga cgtaattaca tccctggggg
     7681 ctttgggggg gggctgtccc tgatatctat aacaagaaaa tatatatata ataagttatc
     7741 acgtaagtag aacatgaaat aacaatataa ttatcgtatg agttaaatct taaaagtcac
     7801 gtaaaagata atcatgcgtc attttgactc acgcggtcgt tatagttcaa aatcagtgac
     7861 acttaccgca ttgacaagca cgcctcacgg gagctccaag cggcgactga gatgtcctaa
     7921 atgcacagcg acggattcgc gctatttaga aagagagagc aatatttcaa gaatgcatgc
     7981 gtcaatttta cgcagactat ctttctaggg ttaatctagc tgcatcagga tcatatcgtc
     8041 gggtcttttt tccggctcag tcatcgccca agctggcgct atctgggcat cggggaggaa
     8101 gaagcccgtg ccttttcccg cgaggttgaa gcggcatgga aagagtttgc cgaggatgac
     8161 tgctgctgca ttgacgttga gcgaaaacgc acgtttacca tgatgattcg ggaaggtgtg
     8221 gccatgcacg cctttaacgg tgaactgttc gttcaggcca cctgggatac cagttcgtcg
     8281 cggcttttcc ggacacagtt ccggatggtc agcccgaagc gcatcagcaa cccgaacaat
     8341 accggcgaca gccggaactg ccgtgccggt gtgcagatta atgacagcgg tgcggcgctg
     8401 ggatattacg tcagcgagga cgggtatcct ggctggatgc cgcagaaatg gacatggata
     8461 ccccgtgagt tacccggcgg gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     8521 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     8581 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     8641 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     8701 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     8761 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     8821 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     8881 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     8941 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     9001 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     9061 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     9121 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     9181 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     9241 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     9301 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     9361 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     9421 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     9481 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     9541 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     9601 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     9661 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     9721 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     9781 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     9841 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     9901 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     9961 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
    10021 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
    10081 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
    10141 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
    10201 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
    10261 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
    10321 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
    10381 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
    10441 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
    10501 cacggaaatg ttgaatactc at
//
