LOCUS       sequence-496442-       14604 bp    DNA     circular SYN 07-JAN-2026
DEFINITION  Overexpression of PPH-T2A-dCas9-NFZ-P2A-BFP with HBB IVS2 intron in 
            PPH coding sequence and blasticidin resistance gene. Contains Cp36 
            recombination site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14604)
  AUTHORS   Arana S, Du PP, Vaughan-Jackson A, Enright N, Spees K, Valbuena R, 
            Garcia CA, Nguyen T, Venida A, Seczynska M, Bintu L, Lehner PJ, 
            Prolo LM, Bassik MC
  TITLE     Reduced Cas9 transgene silencing by incorporation of intron 
            sequences.
  JOURNAL   Nat Commun. 2025 Nov 27;16(1):10656. doi: 
            10.1038/s41467-025-65669-0.
  PUBMED    41309578
REFERENCE   2  (bases 1 to 14604)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 7, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2025-11-27"; volume: "16(1):10656. doi"; pages: " 
            10.1038/s41467-025-65669-0"
COMMENT     Sequence Label: pPD292 PPH/HBB_IVS2/-T2A-dCas9-NFZ-P2A-BFP
FEATURES             Location/Qualifiers
     source          1..14604
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        500..1709
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1546..1565
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             1713..2111
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    2138..2186
                     /note="synthetic polyadenylation signal"
     CDS             join(2452..2520,2521..3312)
                     /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(2670..2689)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3325..3461
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3613..3684
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3689..3735)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3753..3801
                     /note="synthetic polyadenylation signal"
     promoter        3942..5120
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4173..5111
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5068..5088
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(5652..5676)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5726..5746
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7179..7550
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     CDS             7566..7619
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             7623..7670
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(7671..7694,7695..7697,7698..10253,10254..10256,
                     10257..11771)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             11826..11846
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11862..11927
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             12405..12461
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     polyA_signal    13226..13347
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13237..13256)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     13291..13310
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      13462..13508
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      13538..13674
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    13719..13790
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(14016..14604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14096..14115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt
      241 aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat
      301 cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc
      361 tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga
      421 tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata
      481 tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc
      541 ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg
      601 cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc
      661 agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag
      721 agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt
      781 ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt
      841 cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga
      901 gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga
      961 agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac
     1021 tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc
     1081 ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac
     1141 ccaaggtctt gagcccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg
     1201 caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct
     1261 gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg
     1321 cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc
     1381 acctgccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag
     1441 ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc
     1501 gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt
     1561 gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa
     1621 tgtaatcatt tgggtcaata tgtaattttc agtgttagac ttgtaaattg tccgctaaat
     1681 tctggccgtt tttggctttt ttgttagaca acatggccaa gcctttgtct caagaagaat
     1741 ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca
     1801 gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc
     1861 attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag
     1921 ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct
     1981 gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg
     2041 acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt
     2101 gggagggcta agctagtatg taagcctagt cttagataat aaaatcgcta tccatcgaag
     2161 atggatgtgt gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg
     2221 atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat
     2281 ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag
     2341 cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc
     2401 ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt
     2461 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2521 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt
     2581 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc
     2641 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2701 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat
     2761 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2821 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg
     2881 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2941 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3001 atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3061 cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3121 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3181 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3241 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3301 aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg
     3361 cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc
     3421 cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcccgaa tcgtttaaac
     3481 tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc
     3541 atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt tggcagcgat
     3601 cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct ctagtggtca
     3661 taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt ttagaggccc
     3721 caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac acatccatct
     3781 tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccctta accgcttttg
     3841 aatgtccgtc ttagttaatc cctaattgaa cgctccaaaa ggtgacttcc aatagggatt
     3901 tattcctttt aaaattaacg gcataatcgt agatctcgtg aggctccggt gcccgtcagt
     3961 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     4021 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     4081 gcctttttcc cgagggtggg ggagaaccgt atataagtgc actagtcgcc gtgaacgttc
     4141 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
     4201 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt
     4261 acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg tggccttgcg
     4321 cttaaggagc cccttcgcct cgtgcttgag ttgtggcctg gcctgggcgc tggggccgcc
     4381 gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca
     4441 tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg
     4501 cgggccaaga tcagcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc
     4561 gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg
     4621 gacgggggta gtctcaagct gcccggcctg ctctggtgcc tggcctcgcg ccgccgtgta
     4681 tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat
     4741 ggccgcttcc cggccctgct gcagggagca caaaatggag gacgcggcgc tcgggagagc
     4801 gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat
     4861 gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcc agcttttgga
     4921 gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt
     4981 gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc
     5041 tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc
     5101 ttccatttca ggtgtcgtga aaactacccc taaaagccaa aagatctttg tcgatcctac
     5161 catccactcg acacacccgc cagcggccgc tgccaagctt gctagcgcca ccatgtcgcg
     5221 gaggcggcac agctacgaga acgatggtgg acaacctcac aaaaggagga agacgtctat
     5281 gctagcctcc aaaaccatcg ttctttcggt cggcgaggct actcgcactc tgactgagat
     5341 ccagtccacc gcagaccgtc agatcttcga agagaaggtc gggcctctgg tgggtcggct
     5401 gcgcctcacg gcttcgctcc gtcaaaacgg agccaagacc gcgtatcgag tcaacctaaa
     5461 actggatcag gcggacgtcg ttgattccgg acttccgaaa gtgcgctaca ctcaggtatg
     5521 gtcgcacgac gtgacaatcg ttgcgaatag caccgaggcc tcgcgcaaat cgttgtacga
     5581 tttgaccaag tccctcgtcg cgacctcgca ggtcgaagat cttgtcgtca accttgtgcc
     5641 gctgggccgt tcttcaactt tcttgtacaa agtgggcgga ggaggtggaa gcggaggagg
     5701 aggaagcgga ggaggaggta gcggacctaa gaaaaagagg aaggtggcgg ccgctggtac
     5761 cccttcaggg cagatcagca accaggccct ggctctggcc cctagctccg ctccagtgct
     5821 ggcccagact atggtgccct ctagtgctat ggtgcctctg gcccagccac ctgctccagc
     5881 ccctgtgctg accccaggac caccccagtc actgagcgct ccagtgccca agtctgtgag
     5941 tctatgggac gcttgatgtt ttctttcccc ttcttttcta tggttaagtt catgtcatag
     6001 gaaggggata agtaacaggg tacagtttag aatgggaaac agacgaatga ttgcatcagt
     6061 gtggaagtct caggatcgtt ttagtttctt ttatttgctg ttcataacaa ttgttttctt
     6121 ttgtttaatt cttgctttct ttttttttct tctccgcaat ttttactatt atacttaatg
     6181 ccttaacatt gtgtataaca aaaggaaata tctctgagat acattaagta acttaaaaaa
     6241 aaactttaca cagtctgcct agtacattac tatttggaat atatgtgtgc ttatttgcat
     6301 attcataatc tccctacttt attttctttt atttttaatt gatacataat cattatacat
     6361 atttatgggt taaagtgtaa tgttttaata tgtgtacaca tattgaccaa atcagggtaa
     6421 ttttgcattt gtaattttaa aaaatgcttt cttcttttaa tatacttttt tgtttatctt
     6481 atttctaata ctttccctaa tctctttctt tcagggcaat aatgatacaa tgtatcatgc
     6541 ctctttgcac cattctaaag aataacagtg ataatttctg ggttaaggca atagcaatat
     6601 ctctgcatat aaatatttct gcatataaat tgtaactgat gtaagaggtt tcatattgct
     6661 aatagcagct acaatccagc taccattctg cttttatttt atggttggga taaggctgga
     6721 ttattctgag tccaagctag gcccttttgc taatcatgtt catacctctt atcttcctcc
     6781 cacagacaca ggccggcgag gggactctga gtgaagctct gctgcacctg cagttcgacg
     6841 ctgatgagga cctgggagct ctgctgggga acagcaccga tcccggagtg ttcacagatc
     6901 tggcctccgt ggacaactct gagtttcagc agctgctgaa tcagggcgtg tccatgtctc
     6961 atagtacagc cgaaccaatg ctgatggagt accccgaagc cattacccgg ctggtgaccg
     7021 gcagccagcg gccccccgac cccgctccaa ctcccctggg aaccagcggc ctgcctaatg
     7081 ggctgtccgg agatgaagac ttctcaagca tcgctgatat ggactttagt gccctgctgt
     7141 cacagatttc ctctagtggg cagggaggag gtggaagcgg cttcagcgtg gacaccagtg
     7201 ccctgctgga cctgttcagc ccctcggtga ccgtgcccga catgagcctg cctgaccttg
     7261 acagcagcct ggccagtatc caagagctcc tgtctcccca ggagcccccc aggcctcccg
     7321 aggcagagaa cagcagcccg gattcaggga agcagctggt gcactacaca gcgcagccgc
     7381 tgttcctgct ggaccccggc tccgtggaca ccgggagcaa cgacctgccg gtgctgtttg
     7441 agctgggaga gggctcctac ttctccgaag gggacggctt cgccgaggac cccaccatct
     7501 ccctgctgac aggctcggag cctcccaaag ccaaggaccc cactgtctcc gatatcggca
     7561 gtggagaggg cagaggaagt ctgctaacat gcggtgacgt cgaggagaat cctggcccaa
     7621 tgaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag gacaagaagt
     7681 acagcatcgg cctggccatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt
     7741 acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga
     7801 agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga
     7861 agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga
     7921 tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct
     7981 tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg
     8041 aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca
     8101 gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc
     8161 ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt
     8221 tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg
     8281 gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc
     8341 tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggcaacctg attgccctga
     8401 gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc
     8461 agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc
     8521 agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca
     8581 tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat
     8641 acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg
     8701 agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg
     8761 gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg
     8821 gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct
     8881 tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc
     8941 ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga
     9001 ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga
     9061 tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg
     9121 gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg
     9181 agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga
     9241 ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga
     9301 aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga
     9361 aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag
     9421 atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg
     9481 acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac
     9541 tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg
     9601 acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga
     9661 agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt
     9721 ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta
     9781 aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg
     9841 ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg
     9901 acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca
     9961 gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg
    10021 aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc
    10081 agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg
    10141 accaggaact ggacatcaac cggctgtccg actacgatgt ggaccacatc gtgcctcaga
    10201 gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac aaggcccggg
    10261 gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc
    10321 agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga
    10381 gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc
    10441 ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg
    10501 agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg
    10561 atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc
    10621 acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg
    10681 aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga
    10741 gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact
    10801 ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga
    10861 caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga
    10921 aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct
    10981 tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg
    11041 actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg
    11101 tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg
    11161 ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca
    11221 agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg
    11281 agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg
    11341 aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc
    11401 tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact
    11461 acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg
    11521 ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc
    11581 aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca
    11641 agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg
    11701 ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc
    11761 tgggaggcga cagcgctgga ggaggtggaa gcggaggagg aggaagcgga ggaggaggta
    11821 gcggacctaa gaaaaagagg aaggtggcgg ccgctggatc cgactacaaa gaccatgacg
    11881 gtgattataa agatcatgac atcgactaca aggatgacga tgacaagggt ggatccggat
    11941 caggctccga gggccagagc gacgagagag ccctgctgga ccagctgcac accctgctga
    12001 gcaacaccga cgccaccggc ctggaggaga tcgacagagc cctgggcatc cccgagctgg
    12061 tgaaccaggg ccaggccctg gagcccaagc agggaggagg aggtagcgga cacgagaagt
    12121 tccccagcga cctggacctg gacatgttca acggcagcct ggagtgcgac atggagagca
    12181 tcatcagaag cgagctgatg gacgccgacg gcctggactt caacttcgac agcggaggag
    12241 gaggtagcgg attcgtgacc ctgaaggacg tgggcatgga cttcaccctg ggcgactggg
    12301 agcagctggg cctggagcag ggcgacacct tctgggacac cgccctggac aactgccagg
    12361 acctgttcct gctggacccc cccggtggat ccggatcagg ctccgccacg aacttctctc
    12421 tgttaaagca agcaggtgac gtggaagaaa accccggtcc cggatccatg gttagtaagg
    12481 gagaggagtt gataaaagag aatatgcaca tgaaactgta tatggaaggg acagtggaca
    12541 accatcactt caagtgcacc tctgagggag aaggaaagcc atacgaaggc acccagacta
    12601 tgcgaattaa ggtcgtggag ggagggcctc tccccttcgc cttcgatatt cttgcaacat
    12661 ctttcctcta cgggagcaaa accttcatca atcacacaca aggcatacct gactttttta
    12721 agcagagttt tccagagggc tttacctggg agcgtgtgac tacatacgaa gatgggggtg
    12781 tgctgacggc tacacaggat acttcacttc aggacggttg tctgatctac aacgtcaaga
    12841 tcaggggggt caatttcacc tcgaatggac cagtaatgca aaaaaagacc ctgggatggg
    12901 aagcttttac tgaaactctc tatcccgccg acggcggcct cgaaggcaga aacgatatgg
    12961 ctttgaagct ggttggcggg tcccatctaa ttgccaatgc caaaaccaca tataggagca
    13021 aaaagcctgc aaagaacctg aagatgccgg gtgtgtacta tgttgactat agattagagc
    13081 ggatcaaaga ggcaaataac gaaacgtacg tggagcagca cgaggtagcc gtggcgcgct
    13141 actgcgattt accctccaaa ctgggccata aactgaacta acttgactga ctgagataca
    13201 gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca aaccacaact
    13261 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
    13321 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
    13381 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatt
    13441 ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg gtcttgaggg
    13501 gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga aaaaaatgcc
    13561 tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct
    13621 tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt
    13681 cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat ggtcttttct
    13741 ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta atctcattgg
    13801 ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc gagatcgact
    13861 tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg tccttgctat
    13921 tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag
    13981 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
    14041 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
    14101 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
    14161 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
    14221 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
    14281 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
    14341 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
    14401 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
    14461 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
    14521 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
    14581 acgcgcagaa aaaaaggatc tcaa
//
