LOCUS       sequence-364760-       11230 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FKBP12 (F36V mutant) degron-tagged dCas9-KRAB domain fused to 
            tagBFP, expressed under EF1-alpha promoter in a piggyBac plasmid. 
            Expresses Blasticidin resistance under PGK promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11230)
  AUTHORS   Noviello G, Gjaltema RAF, Schulz EG
  TITLE     CasTuner is a degron and CRISPR/Cas-based toolkit for analog tuning 
            of endogenous gene expression.
  JOURNAL   Nat Commun. 2023 Jun 3;14(1):3225. doi: 10.1038/s41467-023-38909-4.
  PUBMED    37270532
REFERENCE   2  (bases 1 to 11230)
  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: "Nat 
            Commun."; date: "2023-06-3"; volume: "14(1):3225. doi"; pages: " 
            10.1038/s41467-023-38909-4"
COMMENT     Sequence Label: 
            pSLPB2B-FKBP12_F36V-SpdCas9-KRAB-tagBFP-PGK-Blasticidin
FEATURES             Location/Qualifiers
     source          1..11230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3..704
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    719..943
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1294..1313
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1369..1767
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    1768..2356
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1821..1841)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2239..2250)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     repeat_region   2539..2601
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2631..2651)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2631..2649)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2670..2686)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2670..2686)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2683..2705)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2694..2710
                     /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(2718..2724,2725..2742,2743..2748))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2763..2784
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2901..2918)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3072..3660)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3152..3171)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3831..4622,4623..4691))
                     /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     4454..4473
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4692..4796)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4822..5277)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4959..4980)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5171..5190
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5404..5426
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5418..5435
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5419..5435
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5445..5464
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5445..5463
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   5479..5513
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        5835..6046
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             6059..6327
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             6379..6399
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6409..6729
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             join(6781..6804,6805..6807,6808..9294,9295..9297,
                     9298..10881)
                     /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 H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6781..10881
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             10882..10902
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10903..10920
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             10999..11184
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
ORIGIN
        1 ccatgagcga gctgattaag gagaacatgc acatgaagct gtacatggag ggcaccgtgg
       61 acaaccatca cttcaagtgc acatccgagg gcgaaggcaa gccctacgag ggcacccaga
      121 ccatgagaat caaggtggtc gagggcggcc ctctcccctt cgccttcgac atcctggcta
      181 ctagcttcct ctacggcagc aagaccttca tcaaccacac ccagggcatc cccgacttct
      241 tcaagcagtc cttccctgag ggcttcacat gggagagagt caccacatac gaagacgggg
      301 gcgtgctgac cgctacccag gacaccagcc tccaggacgg ctgcctcatc tacaacgtca
      361 agatcagagg ggtgaacttc acatccaacg gccctgtgat gcagaagaaa acactcggct
      421 gggaggcctt caccgagacg ctgtaccccg ctgacggcgg cctggaaggc agaaacgaca
      481 tggccctgaa gctcgtgggc gggagccatc tgatcgcaaa catcaagacc acatatagat
      541 ccaagaaacc cgctaagaac ctcaagatgc ctggcgtcta ctatgtggac tacagactgg
      601 aaagaatcaa ggaggccaac aacgagacct acgtcgagca gcacgaggtg gcagtggcca
      661 gatactgcga cctccctagc aaactggggc acaagcttaa ttagccatgg attaacgact
      721 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
      781 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
      841 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
      901 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcctctgg cctcgcacac
      961 attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac
     1021 cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga
     1081 cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc
     1141 aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct
     1201 gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg
     1261 gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt
     1321 ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcaat ggccaagcct
     1381 ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc
     1441 atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact
     1501 ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact
     1561 gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg
     1621 ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc
     1681 aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg
     1741 ccctctggtt atgtgtggga gggctaaaat caacctctgg attacaaaat ttgtgaaaga
     1801 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     1861 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     1921 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     1981 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     2041 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     2101 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     2161 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     2221 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     2281 ccggctctgc ggcctcttcc gcgtctccgc cttcgccctc agacgagtcg gatctccctt
     2341 tgggccgcct ccccgccggt acctaaaagt tttgttactt tatagaagaa attttgagtt
     2401 tttgtttttt tttaataaat aaataaacat aaataaattg tttgttgaat ttattattag
     2461 tatgtaagtg taaatataat aaaacttaat atctattcaa attaataaat aaacctcgat
     2521 atacagaccg ataaaacaca tgcgtcaatt ttacgcatga ttatctttaa cgtacgtcac
     2581 aatatgatta tctttctagg gttaatctag tattaatcca gcttttgttc cctttagtga
     2641 gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     2701 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     2761 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     2821 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     2881 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2941 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3001 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3061 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3121 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3181 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3241 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3301 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3361 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3421 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3481 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     3541 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3601 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3661 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3721 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3781 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3841 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3901 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3961 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4021 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4081 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4141 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4201 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4261 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4321 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4381 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4441 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4501 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4561 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4621 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4681 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4741 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4801 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     4861 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     4921 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     4981 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     5041 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     5101 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     5161 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     5221 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     5281 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     5341 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     5401 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
     5461 agggcgaatt ggggttaacc ctagaaagat agtctgcgta aaattgacgc atgcattctt
     5521 gaaatattgc tctctctttc taaatagcgc gaatccgtcg ctgtgcattt aggacatctc
     5581 agtcgccgct tggagctccc gtgaggcgtg cttgtcaatg cggtaagtgt cactgatttt
     5641 gaactataac gaccgcgtga gtcaaaatga cgcatgatta tcttttacgt gacttttaag
     5701 atttaactca tacgataatt atattgttat ttcatgttct acttacgtga taacttatta
     5761 tatatatatt ttcttgttat agatatcaac tagaatacgc gtaaggatct gcgatcgctc
     5821 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     5881 ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     5941 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     6001 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg
     6061 ggctcgcatc tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt
     6121 gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt
     6181 aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag
     6241 actcagccgg ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc
     6301 gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg gcgcctactc tagagctagc
     6361 gaattcgccg ccaccatgcc taagaaaaag aggaaggtga gcggccgcgg agttcaggtg
     6421 gaaaccatct ccccaggaga cgggcgcacc ttccccaagc gcggccagac ctgcgtggtg
     6481 cactacaccg ggatgcttga agatggaaag aaagttgatt cctcccggga cagaaacaag
     6541 ccctttaagt ttatgctagg caagcaggag gtgatccgag gctgggaaga aggggttgcc
     6601 cagatgagtg tgggtcagag agccaaactg actatatctc cagattatgc ctatggtgcc
     6661 actgggcacc caggcatcat cccaccacat gccactctcg tcttcgatgt ggagcttcta
     6721 aaactggaag gatccggcgg atctggaggt agtggtggat ctggaggttc tggtggctcc
     6781 gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg ggccgtgatc
     6841 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
     6901 agcatcaaga agaacctgat cggcgccctg ctgttcgaca gcggagaaac agccgaggcc
     6961 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
     7021 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
     7081 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
     7141 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
     7201 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
     7261 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
     7321 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
     7381 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
     7441 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggcaacctg
     7501 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
     7561 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
     7621 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
     7681 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
     7741 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
     7801 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
     7861 tacatcgatg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
     7921 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
     7981 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
     8041 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
     8101 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
     8161 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
     8221 gtggacaagg gcgccagcgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
     8281 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtac
     8341 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
     8401 ggcgagcaga aaaaagccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
     8461 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
     8521 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
     8581 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
     8641 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
     8701 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
     8761 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
     8821 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
     8881 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
     8941 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
     9001 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
     9061 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
     9121 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
     9181 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
     9241 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggacgctatc
     9301 gtgcctcaga gctttctgaa ggacgactcc atcgataaca aagtgctgac tcggagcgac
     9361 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
     9421 tactggcgcc agctgctgaa tgccaagctg attacccaga ggaagttcga caatctgacc
     9481 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
     9541 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
     9601 aagtacgacg agaacgacaa actgatccgg gaagtgaaag tgatcaccct gaagtccaag
     9661 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
     9721 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
     9781 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
     9841 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
     9901 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
     9961 ctgatcgaga caaacggcga aacaggcgag atcgtgtggg ataagggccg ggactttgcc
    10021 accgtgcgga aagtgctgtc tatgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
    10081 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgacaa gctgatcgcc
    10141 agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat
    10201 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
    10261 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
    10321 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
    10381 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag
    10441 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
    10501 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
    10561 cacaaacact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
    10621 ctggccgacg ctaatctgga caaggtgctg agcgcctaca acaagcacag agacaagcct
    10681 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
    10741 gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag
    10801 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
    10861 ctgtctcagc tgggaggcga ccccaagaag aagcgcaagg tgcaccacca ccaccaccac
    10921 ggaggaggtg gaagcggagg aggaggaagc ggaggaggag gtagctggat ggatgctaag
    10981 tcactaactg cctggtcccg gacactggtg accttcaagg atgtatttgt ggacttcacc
    11041 agggaggagt ggaagctgct ggacactgct cagcagatcg tgtacagaaa tgtgatgctg
    11101 gagaactata agaacctggt ttccttgggt tatcagctta ctaagccaga tgtgatcctc
    11161 cggttggaga agggagaaga gcccggagga ggtggaagcg gaggaggagg aagcggagga
    11221 ggaggtagcg
//
