LOCUS       sequence-496439-       13754 bp    DNA     circular SYN 07-JAN-2026
DEFINITION  Overexpression of PPH-T2A-dCas9-NFZ-P2A-BFP and blasticidin 
            resistance gene. Contains Cp36 recombination site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13754)
  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 13754)
  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: pPD231 PPH-T2A-dCas9-NFZ-P2A-BFP
FEATURES             Location/Qualifiers
     source          1..13754
                     /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             5762..6304
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     CDS             6329..6700
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     CDS             6716..6769
                     /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             6773..6820
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(6821..6844,6845..6847,6848..9403,9404..9406,
                     9407..10921)
                     /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             10976..10996
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11012..11077
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             11555..11611
                     /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    12376..12497
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12387..12406)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     12441..12460
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      12612..12658
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      12688..12824
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    12869..12940
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(13166..13754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13246..13265)
                     /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 agtctacaca
     5941 ggccggcgag gggactctga gtgaagctct gctgcacctg cagttcgacg ctgatgagga
     6001 cctgggagct ctgctgggga acagcaccga tcccggagtg ttcacagatc tggcctccgt
     6061 ggacaactct gagtttcagc agctgctgaa tcagggcgtg tccatgtctc atagtacagc
     6121 cgaaccaatg ctgatggagt accccgaagc cattacccgg ctggtgaccg gcagccagcg
     6181 gccccccgac cccgctccaa ctcccctggg aaccagcggc ctgcctaatg ggctgtccgg
     6241 agatgaagac ttctcaagca tcgctgatat ggactttagt gccctgctgt cacagatttc
     6301 ctctagtggg cagggaggag gtggaagcgg cttcagcgtg gacaccagtg ccctgctgga
     6361 cctgttcagc ccctcggtga ccgtgcccga catgagcctg cctgaccttg acagcagcct
     6421 ggccagtatc caagagctcc tgtctcccca ggagcccccc aggcctcccg aggcagagaa
     6481 cagcagcccg gattcaggga agcagctggt gcactacaca gcgcagccgc tgttcctgct
     6541 ggaccccggc tccgtggaca ccgggagcaa cgacctgccg gtgctgtttg agctgggaga
     6601 gggctcctac ttctccgaag gggacggctt cgccgaggac cccaccatct ccctgctgac
     6661 aggctcggag cctcccaaag ccaaggaccc cactgtctcc gatatcggca gtggagaggg
     6721 cagaggaagt ctgctaacat gcggtgacgt cgaggagaat cctggcccaa tgaaaaggcc
     6781 ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag gacaagaagt acagcatcgg
     6841 cctggccatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc
     6901 cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat
     6961 cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc
     7021 cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa
     7081 cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga
     7141 agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta
     7201 ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa
     7261 ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt
     7321 cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct
     7381 ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc
     7441 caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca
     7501 gctgcccggc gagaagaaga atggcctgtt cggcaacctg attgccctga gcctgggcct
     7561 gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa
     7621 ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga
     7681 cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt
     7741 gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca
     7801 ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa
     7861 agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag
     7921 ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga
     7981 actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg
     8041 cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga
     8101 tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat
     8161 cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa
     8221 gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc
     8281 ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct
     8341 gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa
     8401 atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat
     8461 cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta
     8521 cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa
     8581 cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga
     8641 caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga
     8701 cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat
     8761 gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa
     8821 cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt
     8881 cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat
     8941 ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc
     9001 cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt
     9061 gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca
     9121 gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat
     9181 caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa
     9241 cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact
     9301 ggacatcaac cggctgtccg actacgatgt ggaccacatc gtgcctcaga gctttctgaa
     9361 ggacgactcc atcgacaaca aggtgctgac cagaagcgac aaggcccggg gcaagagcga
     9421 caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa
     9481 cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct
     9541 gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac
     9601 aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa
     9661 gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa
     9721 ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta
     9781 cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt
     9841 cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga
     9901 aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac
     9961 cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga
    10021 aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag
    10081 catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga
    10141 gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc
    10201 taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa
    10261 agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat
    10321 catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa
    10381 agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa
    10441 cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct
    10501 gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc
    10561 ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga
    10621 gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga
    10681 caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa
    10741 tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga
    10801 caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat
    10861 ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga
    10921 cagcgctgga ggaggtggaa gcggaggagg aggaagcgga ggaggaggta gcggacctaa
    10981 gaaaaagagg aaggtggcgg ccgctggatc cgactacaaa gaccatgacg gtgattataa
    11041 agatcatgac atcgactaca aggatgacga tgacaagggt ggatccggat caggctccga
    11101 gggccagagc gacgagagag ccctgctgga ccagctgcac accctgctga gcaacaccga
    11161 cgccaccggc ctggaggaga tcgacagagc cctgggcatc cccgagctgg tgaaccaggg
    11221 ccaggccctg gagcccaagc agggaggagg aggtagcgga cacgagaagt tccccagcga
    11281 cctggacctg gacatgttca acggcagcct ggagtgcgac atggagagca tcatcagaag
    11341 cgagctgatg gacgccgacg gcctggactt caacttcgac agcggaggag gaggtagcgg
    11401 attcgtgacc ctgaaggacg tgggcatgga cttcaccctg ggcgactggg agcagctggg
    11461 cctggagcag ggcgacacct tctgggacac cgccctggac aactgccagg acctgttcct
    11521 gctggacccc cccggtggat ccggatcagg ctccgccacg aacttctctc tgttaaagca
    11581 agcaggtgac gtggaagaaa accccggtcc cggatccatg gttagtaagg gagaggagtt
    11641 gataaaagag aatatgcaca tgaaactgta tatggaaggg acagtggaca accatcactt
    11701 caagtgcacc tctgagggag aaggaaagcc atacgaaggc acccagacta tgcgaattaa
    11761 ggtcgtggag ggagggcctc tccccttcgc cttcgatatt cttgcaacat ctttcctcta
    11821 cgggagcaaa accttcatca atcacacaca aggcatacct gactttttta agcagagttt
    11881 tccagagggc tttacctggg agcgtgtgac tacatacgaa gatgggggtg tgctgacggc
    11941 tacacaggat acttcacttc aggacggttg tctgatctac aacgtcaaga tcaggggggt
    12001 caatttcacc tcgaatggac cagtaatgca aaaaaagacc ctgggatggg aagcttttac
    12061 tgaaactctc tatcccgccg acggcggcct cgaaggcaga aacgatatgg ctttgaagct
    12121 ggttggcggg tcccatctaa ttgccaatgc caaaaccaca tataggagca aaaagcctgc
    12181 aaagaacctg aagatgccgg gtgtgtacta tgttgactat agattagagc ggatcaaaga
    12241 ggcaaataac gaaacgtacg tggagcagca cgaggtagcc gtggcgcgct actgcgattt
    12301 accctccaaa ctgggccata aactgaacta acttgactga ctgagataca gcgtaccttc
    12361 agctcacaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
    12421 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
    12481 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
    12541 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc
    12601 tatcggtatc gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg
    12661 cccctcgggc cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg
    12721 ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct tccccaactt
    12781 gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct
    12841 gcaggcgatc tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact
    12901 aggggtcaga agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg
    12961 ctatcgaaac ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt
    13021 tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt
    13081 ctccgattac gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc
    13141 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
    13201 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
    13261 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
    13321 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
    13381 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
    13441 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
    13501 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
    13561 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
    13621 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
    13681 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
    13741 aaaaaggatc tcaa
//
