LOCUS       sequence-373487-       11218 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 11218)
  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 11218)
  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..11218
                     /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     1282..1301
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1357..1755
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    1756..2344
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1809..1829)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2227..2238)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     repeat_region   2527..2589
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(2619..2639)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2619..2637)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2658..2674)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2658..2674)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2671..2693)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2682..2698
                     /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(2706..2712,2713..2730,2731..2736))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2751..2772
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2889..2906)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3060..3648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3140..3159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3819..4610,4611..4679))
                     /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     4442..4461
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4680..4784)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4810..5265)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4947..4968)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5159..5178
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5392..5414
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5406..5423
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5407..5423
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5433..5452
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5433..5451
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   5467..5501
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        5823..6034
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             6047..6315
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             6367..6387
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6397..6717
                     /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(6769..6792,6793..6795,6796..9282,9283..9285,
                     9286..10869)
                     /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             6769..10869
                     /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             10870..10890
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10891..10908
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             10987..11172
                     /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 gcgggcgccc
     1261 gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt
     1321 tctcctcttc ctcatctccg ggcctttcga cctgcaatgg ccaagccttt gtctcaagaa
     1381 gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac
     1441 tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta
     1501 tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg
     1561 gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc
     1621 ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg
     1681 aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat
     1741 gtgtgggagg gctaaaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     1801 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     1861 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     1921 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     1981 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     2041 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     2101 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     2161 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     2221 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     2281 cctcttccgc gtctccgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     2341 ccgccggtac ctaaaagttt tgttacttta tagaagaaat tttgagtttt tgtttttttt
     2401 taataaataa ataaacataa ataaattgtt tgttgaattt attattagta tgtaagtgta
     2461 aatataataa aacttaatat ctattcaaat taataaataa acctcgatat acagaccgat
     2521 aaaacacatg cgtcaatttt acgcatgatt atctttaacg tacgtcacaa tatgattatc
     2581 tttctagggt taatctagta ttaatccagc ttttgttccc tttagtgagg gttaattgcg
     2641 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2701 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2761 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2821 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2881 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2941 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3001 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3061 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3121 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3181 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3241 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3301 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3361 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3421 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3481 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3541 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3601 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3661 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3721 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3781 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3841 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3901 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3961 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4021 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4081 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4141 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4201 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4261 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4321 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4381 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4441 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4501 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4561 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4621 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4681 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4741 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4801 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     4861 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     4921 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     4981 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     5041 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     5101 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     5161 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     5221 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca
     5281 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     5341 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     5401 gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg
     5461 ggctaaccct agaaagatag tctgcgtaaa attgacgcat gcattcttga aatattgctc
     5521 tctctttcta aatagcgcga atccgtcgct gtgcatttag gacatctcag tcgccgcttg
     5581 gagctcccgt gaggcgtgct tgtcaatgcg gtaagtgtca ctgattttga actataacga
     5641 ccgcgtgagt caaaatgacg catgattatc ttttacgtga cttttaagat ttaactcata
     5701 cgataattat attgttattt catgttctac ttacgtgata acttattata tatatatttt
     5761 cttgttatag atatcaacta gaatacgcgt aaggatctgc gatcgctccg gtgcccgtca
     5821 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
     5881 aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
     5941 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
     6001 tctttttcgc aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc
     6061 tccttcacgc gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct
     6121 gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct
     6181 caggtcgaga ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct
     6241 ctccacgctt tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct
     6301 gcgccgttac agatccaagc tgtgaccggc gcctactcta gagctagcga attcgccgcc
     6361 accatgccta agaaaaagag gaaggtgagc ggccgcggag ttcaggtgga aaccatctcc
     6421 ccaggagacg ggcgcacctt ccccaagcgc ggccagacct gcgtggtgca ctacaccggg
     6481 atgcttgaag atggaaagaa agttgattcc tcccgggaca gaaacaagcc ctttaagttt
     6541 atgctaggca agcaggaggt gatccgaggc tgggaagaag gggttgccca gatgagtgtg
     6601 ggtcagagag ccaaactgac tatatctcca gattatgcct atggtgccac tgggcaccca
     6661 ggcatcatcc caccacatgc cactctcgtc ttcgatgtgg agcttctaaa actggaagga
     6721 tccggcggat ctggaggtag tggtggatct ggaggttctg gtggctccga caagaagtac
     6781 agcatcggcc tggccatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac
     6841 aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag
     6901 aacctgatcg gcgccctgct gttcgacagc ggagaaacag ccgaggccac ccggctgaag
     6961 agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc
     7021 ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc
     7081 ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag
     7141 gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc
     7201 accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg
     7261 ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc
     7321 atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc
     7381 gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg
     7441 atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gcaacctgat tgccctgagc
     7501 ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag
     7561 ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag
     7621 tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc
     7681 ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac
     7741 gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag
     7801 aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta catcgatggc
     7861 ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc
     7921 accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc
     7981 gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg
     8041 caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc
     8101 ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg
     8161 accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc
     8221 gccagcgccc agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag
     8281 aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtacaa cgagctgacc
     8341 aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa
     8401 aaagccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa
     8461 gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat
     8521 cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac
     8581 ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg
     8641 tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac
     8701 aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag
     8761 ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc
     8821 gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa
     8881 gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc
     8941 aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac
     9001 gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga
     9061 gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa
     9121 gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag
     9181 ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac
     9241 caggaactgg acatcaaccg gctgtccgac tacgatgtgg acgctatcgt gcctcagagc
     9301 tttctgaagg acgactccat cgataacaaa gtgctgactc ggagcgacaa gaaccggggc
     9361 aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcgccag
     9421 ctgctgaatg ccaagctgat tacccagagg aagttcgaca atctgaccaa ggccgagaga
     9481 ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg
     9541 cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag
     9601 aacgacaaac tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat
     9661 ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac
     9721 gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa
     9781 agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc
     9841 gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt
     9901 ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca
     9961 aacggcgaaa caggcgagat cgtgtgggat aagggccggg actttgccac cgtgcggaaa
    10021 gtgctgtcta tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc
    10081 agcaaagagt ctatcctgcc caagaggaac agcgacaagc tgatcgccag aaagaaggac
    10141 tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg
    10201 gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg
    10261 atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag
    10321 ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag
    10381 ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa
    10441 ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg
    10501 aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca caaacactac
    10561 ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct
    10621 aatctggaca aggtgctgag cgcctacaac aagcacagag acaagcctat cagagagcag
    10681 gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag
    10741 tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc
    10801 accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg
    10861 ggaggcgacc ccaagaagaa gcgcaaggtg caccaccacc accaccacgg aggaggtgga
    10921 agcggaggag gaggaagcgg aggaggaggt agctggatgg atgctaagtc actaactgcc
    10981 tggtcccgga cactggtgac cttcaaggat gtatttgtgg acttcaccag ggaggagtgg
    11041 aagctgctgg acactgctca gcagatcgtg tacagaaatg tgatgctgga gaactataag
    11101 aacctggttt ccttgggtta tcagcttact aagccagatg tgatcctccg gttggagaag
    11161 ggagaagagc ccggaggagg tggaagcgga ggaggaggaa gcggaggagg aggtagcg
//
