LOCUS       sequence-410725-        9699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac transposon vector suitable for inserting the ChR2-YFP 
            optogenetic actuator driven by the ubiquitous CAG promoter into the 
            genome of mammalian cell lines..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9699)
  AUTHORS   Harley P, Kerins C, Gatt A, Neves G, Riccio F, Machado CB, 
            Cheesbrough A, R'Bibo L, Burrone J, Lieberam I
  TITLE     Aberrant axon initial segment plasticity and intrinsic excitability 
            of ALS hiPSC motor neurons.
  JOURNAL   Cell Rep. 2023 Dec 26;42(12):113509. doi: 
            10.1016/j.celrep.2023.113509. Epub 2023 Nov 28.
  PUBMED    38019651
REFERENCE   2  (bases 1 to 9699)
  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: "Cell Rep.";
            date: "2023-12-26"; volume: "42(12):113509. doi"; pages: " 
            10.1016/j.celrep.2023.113509. Epub 2023 Nov 28"
COMMENT     Sequence Label: PB-CAG-ChR2-YFP
FEATURES             Location/Qualifiers
     source          1..9699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     266..285
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     519..536
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    653..674
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(689..694,695..712,713..719)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    727..743
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     732..754
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     751..767
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     751..767
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   1258..1292
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    complement(1588..1635)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     polyA_signal    1664..1888
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(1924..2943)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             complement(1924..2943)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     promoter        complement(2977..3024)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     primer_bind     complement(3089..3108)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     3482..3505
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    complement(3529..3576)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     polyA_signal    3638..3862
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(join(3941..4654,4655..4657,4658..4660))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3986..4007)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4335..4354
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4594..4615
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4657..4666
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4676..5602)
                     /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"
     primer_bind     complement(5659..5678)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          5686..6706
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(5744..5766)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     enhancer        6943..7246
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     repeat_region   7509..7571
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7701..7718)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7701..7717)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7710..7732)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7859..8314
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7946..7965)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8156..8177
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8340..8444
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8445..8513,8514..9305)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8663..8682)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(9476..9699,1..365)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
       61 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      121 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      181 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      241 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      301 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      361 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      421 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      481 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      541 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
      601 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
      661 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
      721 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
      781 agcgcgcccg ccgggtaact cacggggtat ccatgtccat ttctgcggca tccagccagg
      841 atacccgtcc tcgctgacgt aatatcccag cgccgcaccg ctgtcattaa tctgcacacc
      901 ggcacggcag ttccggctgt cgccggtatt gttcgggttg ctgatgcgct tcgggctgac
      961 catccggaac tgtgtccgga aaagccgcga cgaactggta tcccaggtgg cctgaacgaa
     1021 cagttcaccg ttaaaggcgt gcatggccac accttcccga atcatcatgg taaacgtgcg
     1081 ttttcgctca acgtcaatgc agcagcagtc atcctcggca aactctttcc atgccgcttc
     1141 aacctcgcgg gaaaaggcac gggcttcttc ctccccgatg cccagatagc gccagcttgg
     1201 gcgatgactg agccggaaaa aagacccgac gatatgatcc tgatgcagct agattaaccc
     1261 tagaaagata gtctgcgtaa aattgacgca tgcattcttg aaatattgct ctctctttct
     1321 aaatagcgcg aatccgtcgc tgtgcattta ggacatctca gtcgccgctt ggagctcccg
     1381 tgaggcgtgc ttgtcaatgc ggtaagtgtc actgattttg aactataacg accgcgtgag
     1441 tcaaaatgac gcatgattat cttttacgtg acttttaaga tttaactcat acgataatta
     1501 tattgttatt tcatgttcta cttacgtgat aacttattat atatatattt tcttgttata
     1561 gatatcaagc ttatcgatac cgtcgacgaa gttcctatac tttctagaga ataggaactt
     1621 cggaatagga acttctccat ggtagccctt tccgcctcag aagccataga gcccaccgca
     1681 tccccagcat gcctgctatt gtcttcccaa tcctccccct tgctgtcctg ccccacccca
     1741 ccccccagaa tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag
     1801 gaaaggacag tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca
     1861 acagatggct ggcaactaga aggcacagtc gaggcgtcag cgagctctag agaattgatc
     1921 cccctattcc tttgccctcg gacgagtgct ggggcgtcgg tttccactat cggcgagtac
     1981 ttctacacag ccatcggtcc agacggccgc gcttctgcgg gcgatttgtg tacgcccgac
     2041 agtcccggct ccggatcgga cgattgcgtc gcatcgaccc tgcgcccaag ctgcatcatc
     2101 gaaattgccg tcaaccaagc tctgatagag ttggtcaaga ccaatgcgga gcatatacgc
     2161 ccggagccgc ggcgatcctg caagctccgg atgcctccgc tcgaagtagc gcgtctgctg
     2221 ctccatacaa gccaaccacg gcctccagaa gaagatgttg gcgacctcgt attgggaatc
     2281 cccgaacatc gcctcgctcc agtcaatgac cgctgttatg cggccattgt ccgtcaggac
     2341 attgttggag ccgaaatccg cgtgcacgag gtgccggact tcggggcagt cctcggccca
     2401 aagcatcagc tcatcgagag cctgcgcgac ggacgcactg acggtgtcgt ccatcacagt
     2461 ttgccagtga tacacatggg gatcagcaat cgcgcatatg aaatcacgcc atgtagtgta
     2521 ttgaccgatt ccttgcggtc cgaatgggcc gaacccgctc gtctggctaa gatcggccgc
     2581 agcgatcgca tccatggcct ccgcgaccgg ctgcagaaca gcgggcagtt cggtttcagg
     2641 caggtcttgc aacgtgacac cctgtgcacg gcgggagatg caataggtca ggctctcgct
     2701 gaattcccca atgtcaagca cttccggaat cgggagcgcg gccgatgcaa agtgccgata
     2761 aacataacga tctttgtaga aaccatcggc gcagctattt acccgcagga catatccacg
     2821 ccctcctaca tcgaagctga aagcacgaga ttcttcgccc tccgagagct gcatcaggtc
     2881 ggagacgctg tcgaactttt cgatcagaaa cttctcgaca gacgtcgcgg tgagttcagg
     2941 ctttccggat ctatccatgg tttagttcct caccttgtcg tattatacta tgccgatata
     3001 ctatgccgat gattaattgt caacacgtgc tgctgcaggt cgaaaggccc ggagatgagg
     3061 aagaggagaa cagcgcggca gacgtgcgct tttgaagcgt gcagaatgcc gggcctccgg
     3121 aggaccttcg ggcgcccgcc ccgcccctga gcccgcccct gagcccgccc ccggacccac
     3181 cccttcccag cctctgagcc cagaaagcga aggagcaaag ctgctattgg ccgctgcccc
     3241 aaaggcctac ccgcttccat tgctcagcgg tgctgtccat ctgcacgaga ctagtgagac
     3301 gtgctacttc catttgtcac gtcctgcacg acgcgagctg cggggcgggg gggaacttcc
     3361 tgactagggg aggagtagaa ggtggcgcga aggggccacc aaagaacgga gccggttggc
     3421 gcctaccggt ggatgtggaa tgtgtgcgag gccagaggcc acttgtgtag cgccaagtgc
     3481 ccagcggggc tgctaaagcg catgctccag actgccttgg gaaaagcgga agttcctata
     3541 ctttctagag aataggaact tcggaatagg aacttctcca tggtagccct taattaaact
     3601 agactcgagc cccagctggt tctttccgcc tcagaagcca tagagcccac cgcatcccca
     3661 gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc
     3721 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
     3781 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
     3841 ggctggcaac tagaaggcac agtcgaggct gatcagcgag ctctagggct agcgggcccc
     3901 catgggatcc actagtggat cccccgggct gcaggaattc ttacttgtac agctcgtcca
     3961 tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga tcgcgcttct
     4021 cgttggggtc tttgctcagg gcggactggt agctcaggta gtggttgtcg ggcagcagca
     4081 cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt
     4141 cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca
     4201 tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg atgttgccgt
     4261 cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg ccctcgaact
     4321 tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg cgctcctgga
     4381 cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc
     4441 gggcgaagca ctgcaggccg tagccgaagg tggtcacgag ggtgggccag ggcacgggca
     4501 gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc
     4561 cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg
     4621 gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtggcggcc gctggcacgg
     4681 ctccggcctc ggcttcgtct tcgacgagag tctcgacctc gatctccgtt ccgccgatgt
     4741 tcagtttggt ggttttgcgg atatctccgt gaatcaatat gtgctcgtgg atcaggacgc
     4801 gcaggtagtg tcccaacaac ccccaacaat ttttactcat cagatcaata atcgtgtgac
     4861 ctacggtgga gccatagacg ctcaggacgc caaaaccttc gggccccaaa atgaagagaa
     4921 ttgggaacat accccagctc acgaaaaaca gccatgccat gccggtcacg acctggcggc
     4981 accgaccctt tggcacagta tgataaccct cgatatatgc tttggcggcg tgaaaaaatg
     5041 tgttcgcgcc atagcacaat ccaagacaaa agaagatgac tttaacatag ccggttgcca
     5101 tggcgctggt agccccccac acgatagtcc cgatgtctga gacaaggagt cccatggttc
     5161 tcctgctgta gtcgttgctc aggccggtga ggttgctcag gcggataagg atgacaggac
     5221 aagtgagcag ccactctgca tagcgcagcc actgcacccg gtgtcctgtg gcaaggtaga
     5281 gcatagaggg attcttaaac tcaaaaaaga actcgagaat caccttaacc atttcaatgg
     5341 cgcacacata gatctcctcc cagccgcatg tagatttcca ggtttggtag gcatagaaca
     5401 tcagcagcaa aatgctgaat cctgctgcaa gccactgcag gacatttgac gcggtctgag
     5461 cgccgttcgt gccgcgagat tcaatccatc cggcacagta acattgatcc tcagggacca
     5521 ggacggaccc gttcaccacc acaggattag taacgaacaa aagttcgcgt ccgacggcag
     5581 acaaagcgcc gccatagtcc atggtggacc ggtaagtgac agtggcgcgc cgaattcttt
     5641 gccaaaatga tgagacagca caataaccag cacgttgccc aggagctgta ggaaaaagaa
     5701 gaaggcatga acatggttag cagaggctct agctagagcc gccggtcaca cgccagaagc
     5761 cgaaccccgc cctgccccgt cccccccgaa ggcagccgtc cccccgcgga cagccccgag
     5821 gctggagagg gagaagggga cggcggcgcg gcgacgcacg aaggccctcc ccgcccattt
     5881 ccttcctgcc ggcgccgcac cgcttcgccc cgcgcccgct agagggggtg cggcggcgcc
     5941 tcccagattt cggctccgca cagatttggg acaaaggaag tccctgcgcc ctctcgcacg
     6001 attaccataa aaggcaatgg ctgcggctcg ccgcgcctcg acagccgccg gcgctccggg
     6061 ggccgccgcg cccctccccc gagccctccc cggcccgagg cggccccgcc ccgcccggca
     6121 cccccacctg ccgccacccc ccgcccggca cggcgagccc cgcgccacgc cccgtacgga
     6181 gccccgcacc cgaagccggg ccgtgctcag caactcgggg aggggggtgc agggggggtt
     6241 gcagcccgac cgacgcgccc acaccccctg ctcacccccc cacgcacaca ccccgcacgc
     6301 agcctttgtt cccctcgcag ccccccccgc accgcggggc accgcccccg gccgcgctcc
     6361 cctcgcgcac actgcggagc gcacaaagcc ccgcgccgcg cccgcagcgc tcacagccgc
     6421 cgggcagcgc ggagccgcac gcggcgctcc ccacgcacac acacacgcac gcaccccccg
     6481 agccgctccc cccgcacaaa gggccctccc ggagcccctc aaggctttca cgcagccaca
     6541 gaaaagaaac aagccgtcat taaaccaagc gctaattaca gcccggagga gaagggccgt
     6601 cccgcccgct cacctgtggg agtaacgcgg tcagtcagag ccggggcggg cggcgcgagg
     6661 cggcggcgga gcggggcacg gggcgaaggc agcgcgcagc gactcccgcc cgccgcgcgc
     6721 ttcgcttttt atagggccgc cgccgccgcc gcctcgccat aaaaggaaac tttcggagcg
     6781 cgccgctctg attggctgcc gccgcacctc tccgcctcgc cccgccccgc cccycgcccc
     6841 smycssyscm smaaataatt aaaaaataaa taaatacaaa attgggggtg gggagggggg
     6901 ggagatgggg agagtgaagc agaacgtggg gctcacctcg accatggtaa tagcgatgac
     6961 taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg
     7021 ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac
     7081 ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa
     7141 agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc
     7201 gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata
     7261 tgggctatga actaatgacc ccgtaattga ttactattaa taactagtgg atcccccggg
     7321 ctgcaggaat tcgataaaag ttttgttact ttatagaaga aattttgagt ttttgttttt
     7381 tttaataaat aaataaacat aaataaattg tttgttgaat ttattattag tatgtaagtg
     7441 taaatataat aaaacttaat atctattcaa attaataaat aaacctcgat atacagaccg
     7501 ataaaacaca tgcgtcaatt ttacacatga ttatctttaa cgtacgtcac aatatgatta
     7561 tctttctagg gttaatctag ctgcgtgttc tgcagcgtgt cgagcatctt catctgctcc
     7621 atcacgctgt aaaacacatt tgcaccgcga gtctgcccgt cctccacggg ttcaaaaacg
     7681 tgaatgaacg aggcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     7741 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     7801 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     7861 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     7921 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     7981 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     8041 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     8101 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     8161 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     8221 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     8281 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     8341 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     8401 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     8461 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     8521 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     8581 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     8641 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     8701 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     8761 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     8821 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     8881 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     8941 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     9001 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     9061 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     9121 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     9181 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     9241 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     9301 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     9361 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     9421 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     9481 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     9541 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     9601 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     9661 actctgtagc accgcctaca tacctcgctc tgctaatcc
//
