LOCUS       sequence-392604-       11541 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAGp-Target-AID-2A-BlastR for PiggyBac genomic integration.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11541)
  AUTHORS   Ishiguro S, Ishida K, Sakata RC, Ichiraku M, Takimoto R, Yogo R, 
            Kijima Y, Mori H, Tanaka M, King S, Tarumoto S, Tsujimura T, Bashth 
            O, Masuyama N, Adel A, Toyoshima H, Seki M, Oh JH, Archambault AS, 
            Nishida K, Kondo A, Kuhara S, Aburatani H, Klein Geltink RI, 
            Yamamoto T, Shakiba N, Takashima Y, Yachie N
  TITLE     A multi-kingdom genetic barcoding system for precise clone 
            isolation.
  JOURNAL   Nat Biotechnol. 2025 May 21. doi: 10.1038/s41587-025-02649-1.
  PUBMED    40399693
REFERENCE   2  (bases 1 to 11541)
  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 
            Biotechnol. 2025 May 21. doi: 10.1038/s41587-025-02649-1."
COMMENT     Sequence Label: pNM1325
FEATURES             Location/Qualifiers
     source          1..11541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2739..2759
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2967..3032
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3666..3686
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3693..3944
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             3945..3998
                     /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             4005..4403
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(4431..4448)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4437..4661
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4708..4724)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4732..4748
                     /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(4756..4762,4763..4780,4781..4786))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4801..4822
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   5056..5118
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5159..5179)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5159..5177)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5198..5214)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5222..5238
                     /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(5246..5252,5253..5270,5271..5276))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5291..5312
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5429..5446)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5600..6188)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5680..5699)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6359..7150,7151..7219))
                     /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     6982..7001
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7220..7324)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7350..7805)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7487..7508)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7699..7718
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7932..7954
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7946..7963
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7947..7963
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7973..7992
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7973..7991
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   8021..8055
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        8358..8737
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          9015..10031
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     9955..9977
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     10039..10058
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             10119..10139
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(10164..10190,10191..10193,10194..11541,1..2726)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(10164..11541,1..2726)
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
ORIGIN
        1 ccagattcgc gtggatgact cgcaaatcag aagagaccat cactccctgg aacttcgagg
       61 aagtcgtgga taagggggcc tctgcccagt ccttcatcga aaggatgact aactttgata
      121 aaaatctgcc taacgaaaag gtgcttccta aacactctct gctgtacgag tacttcacag
      181 tttataacga gctcaccaag gtcaaatacg tcacagaagg gatgagaaag ccagcattcc
      241 tgtctggaga gcagaagaaa gctatcgtgg acctcctctt caagacgaac cggaaagtta
      301 ccgtgaaaca gctcaaagaa gactatttca aaaagattga atgtttcgac tctgttgaaa
      361 tcagcggagt ggaggatcgc ttcaacgcat ccctgggaac gtatcacgat ctcctgaaaa
      421 tcattaaaga caaggacttc ctggacaatg aggagaacga ggacattctt gaggacattg
      481 tcctcaccct tacgttgttt gaagataggg agatgattga agaacgcttg aaaacttacg
      541 ctcatctctt cgacgacaaa gtcatgaaac agctcaagag gcgccgatat acaggatggg
      601 ggcggctgtc aagaaaactg atcaatggga tccgagacaa gcagagtgga aagacaatcc
      661 tggattttct taagtccgat ggatttgcca accggaactt catgcagttg atccatgatg
      721 actctctcac ctttaaggag gacatccaga aagcacaagt ttctggccag ggggacagtc
      781 ttcacgagca catcgctaat cttgcaggta gcccagctat caaaaaggga atactgcaga
      841 ccgttaaggt cgtggatgaa ctcgtcaaag taatgggaag gcataagccc gagaatatcg
      901 ttatcgagat ggcccgagag aaccaaacta cccagaaggg acagaagaac agtagggaaa
      961 ggatgaagag gattgaagag ggtataaaag aactggggtc ccaaatcctt aaggaacacc
     1021 cagttgaaaa cacccagctt cagaatgaga agctctacct gtactacctg cagaacggca
     1081 gggacatgta cgtggatcag gaactggaca tcaatcggct ctccgactac gacgtggatc
     1141 atatcgtgcc ccagtctttt ctcaaagatg attctattga taataaagtg ttgacaagat
     1201 ccgataaaaa tagagggaag agtgataacg tcccctcaga agaagttgtc aagaaaatga
     1261 aaaattattg gcggcagctg ctgaacgcca aactgatcac acaacggaag ttcgataatc
     1321 tgactaaggc tgaacgaggt ggcctgtctg agttggataa agccggcttc atcaaaaggc
     1381 agcttgttga gacacgccag atcaccaagc acgtggccca aattctcgat tcacgcatga
     1441 acaccaagta cgatgaaaat gacaaactga ttcgagaggt gaaagttatt actctgaagt
     1501 ctaagctggt ctcagatttc agaaaggact ttcagtttta taaggtgaga gagatcaaca
     1561 attaccacca tgcgcatgat gcctacctga atgcagtggt aggcactgca cttatcaaaa
     1621 aatatcccaa gcttgaatct gaatttgttt acggagacta taaagtgtac gatgttagga
     1681 aaatgatcgc aaagtctgag caggaaatag gcaaggccac cgctaagtac ttcttttaca
     1741 gcaatattat gaattttttc aagaccgaga ttacactggc caatggagag attcggaagc
     1801 gaccacttat cgaaacaaac ggagaaacag gagaaatcgt gtgggacaag ggtagggatt
     1861 tcgcgacagt ccggaaggtc ctgtccatgc cgcaggtgaa catcgttaaa aagaccgaag
     1921 tacagaccgg aggcttctcc aaggaaagta tcctcccgaa aaggaacagc gacaagctga
     1981 tcgcacgcaa aaaagattgg gaccccaaga aatacggcgg attcgattct cctacagtcg
     2041 cttacagtgt actggttgtg gccaaagtgg agaaagggaa gtctaaaaaa ctcaaaagcg
     2101 tcaaggaact gctgggcatc acaatcatgg agcgatcaag cttcgaaaaa aaccccatcg
     2161 actttctcga ggcgaaagga tataaagagg tcaaaaaaga cctcatcatt aagcttccca
     2221 agtactctct ctttgagctt gaaaacggcc ggaaacgaat gctcgctagt gcgggcgagc
     2281 tgcagaaagg taacgagctg gcactgccct ctaaatacgt taatttcttg tatctggcca
     2341 gccactatga aaagctcaaa gggtctcccg aagataatga gcagaaacag ctgttcgtgg
     2401 aacaacacaa acactacctt gatgagatca tcgagcaaat aagcgaattc tccaaaagag
     2461 tgatcctcgc cgacgctaac ctcgataagg tgctttctgc ttacaataag cacagggata
     2521 agcccatcag ggagcaggca gaaaacatta tccacttgtt tactctgacc aacttgggcg
     2581 cgcctgcagc cttcaagtac ttcgacacca ccatagacag aaagcggtac acctctacaa
     2641 aggaggtcct ggacgccaca ctgattcatc agtcaattac ggggctctat gaaacaagaa
     2701 tcgacctctc tcagctcggt ggagacagca gggctgaccc caagaagaag aggaaggtgg
     2761 gtggaggagg taccggcggt ggaggctcag cagaatacgt acgagctctg tttgacttca
     2821 atgggaatga cgaggaggat ctccccttta agaagggcga tattctccgc atcagagata
     2881 agcccgaaga acaatggtgg aatgccgagg atagcgaagg gaaaaggggc atgattctgg
     2941 tgccatatgt ggagaaatat tccggtgact acaaagacca tgatggggat tacaaagacc
     3001 acgacatcga ctacaaagac gacgacgata aatcagggat gacagacgcc gagtacgtgc
     3061 gcattcatga gaaactggat atttacacct tcaagaagca gttcttcaac aacaagaaat
     3121 ctgtgtcaca ccgctgctac gtgctgtttg agttgaagcg aaggggcgaa agaagggctt
     3181 gcttttgggg ctatgccgtc aacaagcccc aaagtggcac cgagagagga atacacgctg
     3241 agatattcag tatccgaaag gtggaagagt atcttcggga taatcctggg cagtttacga
     3301 tcaactggta ttccagctgg agtccttgcg ctgattgtgc cgagaaaatt ctggaatggt
     3361 ataatcagga acttcgggga aacgggcaca cattgaaaat ctgggcctgc aagctgtact
     3421 acgagaagaa tgcccggaac cagataggac tctggaatct gagggacaat ggtgtaggcc
     3481 tgaacgtgat ggtttccgag cactatcagt gttgtcggaa gattttcatc caaagctctc
     3541 ataaccagct caatgaaaac cgctggttgg agaaaacact gaaacgtgcg gagaagtgga
     3601 gatccgagct gagcatcatg atccaggtca agattctgca taccactaag tctccagccg
     3661 ttggtcccaa gaagaaaaga aaagtcggta ccatgaccaa cctttccgac atcatagaga
     3721 aggaaacagg caaacagttg gtcatccaag agtcgatact catgcttcct gaagaagttg
     3781 aggaggtcat tgggaataag ccggaaagtg acattctcgt acacactgcg tatgatgaga
     3841 gcaccgatga gaacgtgatg ctgctcacgt cagatgcccc agagtacaaa ccctgggctc
     3901 tggtgattca ggactctaat ggagagaaca agatcaagat gctagaaggc aggggaagcc
     3961 ttctgacttg tggggatgtg gaagaaaacc ctggtccatc tagaatggcc aagcctttgt
     4021 ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc atccccatct
     4081 ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg
     4141 tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg
     4201 ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca
     4261 tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag
     4321 ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct
     4381 ctggttatgt gtgggagggc taaatcgatg atctagagct cgctgatcag cctcgactgt
     4441 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     4501 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     4561 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     4621 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     4681 ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4741 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4801 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4861 cctgtcgtgc cagcggatct taaaagtttt gttactttat agaagaaatt ttgagttttt
     4921 gttttttttt aataaataaa taaacataaa taaattgttt gttgaattta ttattagtat
     4981 gtaagtgtaa atataataaa acttaatatc tattcaaatt aataaataaa cctcgatata
     5041 cagaccgata aaacacatgc gtcaatttta cgcatgatta tctttaacgt acgtcacaat
     5101 atgattatct ttctagggtt aatctagtat acgcgtttaa ttaatccagc ttttgttccc
     5161 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     5221 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5281 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5341 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5401 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5461 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5521 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5581 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5641 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5701 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5761 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5821 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5881 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5941 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6001 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6061 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6121 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6181 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6241 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6301 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6361 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6421 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6481 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6541 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6601 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6661 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6721 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6781 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6841 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6901 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6961 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7021 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7081 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7141 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7201 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7261 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7321 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     7381 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     7441 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     7501 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     7561 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     7621 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     7681 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     7741 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     7801 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     7861 ttcgctatta cgccagctgg cgaaagggag atgtgctgca aggcgattaa gttgggtaac
     7921 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga
     7981 ctcactatag ggcgaattgg ggcgcgccat tctagattaa ccctagaaag atagtctgcg
     8041 taaaattgac gcatgcattc ttgaaatatt gctctctctt tctaaatagc gcgaatccgt
     8101 cgctgtgcat ttaggacatc tcagtcgccg cttggagctc ccgtgaggcg tgcttgtcaa
     8161 tgcggtaagt gtcactgatt ttgaactata acgaccgcgt gagtcaaaat gacgcatgat
     8221 tatcttttac gtgactttta agatttaact catacgataa ttatattgtt atttcatgtt
     8281 ctacttacgt gataacttat tatatatata ttttcttgtt atagatatca actagaatgc
     8341 tagcatgggc ccatctcgac attgattatt gactagttat taatagtaat caattacggg
     8401 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     8461 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     8521 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     8581 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     8641 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     8701 gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg
     8761 cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta
     8821 attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgccrggs gggggggggs
     8881 ggggsgrggg gsggggsggg gsgrggcgga gaggkgcggc ggcagccaat cagagcggcg
     8941 cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa
     9001 gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg
     9061 cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg
     9121 acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc
     9181 tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct
     9241 cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg
     9301 gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg
     9361 ggagcgcggc cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct
     9421 gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc
     9481 aaccccccct gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc
     9541 tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg
     9601 tgccgggcgg ggcggggccg cctcgggccg gggrgggctc gggggagggg cgcggcggcc
     9661 cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat
     9721 cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga
     9781 ggcgccgccg caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg
     9841 aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc
     9901 ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg
     9961 gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct
    10021 ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag
    10081 aattgggatc cgaattcctc gagggcggcc gcatggcacc gaagaagaag cgtaaagtcg
    10141 gaatccacgg agttcctgcg gcaatggaca agaagtactc cattgggctc gctatcggca
    10201 caaacagcgt cggttgggcc gtcattacgg acgagtacaa ggtgccgagc aaaaaattca
    10261 aagttctggg caataccgat cgccacagca taaagaagaa cctcattggc gccctcctgt
    10321 tcgactccgg ggagacggcc gaagccacgc ggctcaaaag aacagcacgg cgcagatata
    10381 cccgcagaaa gaatcggatc tgctacctgc aggagatctt tagtaatgag atggctaagg
    10441 tggatgactc tttcttccat aggctggagg agtccttttt ggtggaggag gataaaaagc
    10501 acgagcgcca cccaatcttt ggcaatatcg tggacgaggt ggcgtaccat gaaaagtacc
    10561 caaccatata tcatctgagg aagaagcttg tagacagtac tgataaggct gacttgcggt
    10621 tgatctatct cgcgctggcg catatgatca aatttcgggg acacttcctc atcgaggggg
    10681 acctgaaccc agacaacagc gatgtcgaca aactctttat ccaactggtt cagacttaca
    10741 atcagctttt cgaagagaac ccgatcaacg catccggagt tgacgccaaa gcaatcctga
    10801 gcgctaggct gtccaaatcc cggcggctcg aaaacctcat cgcacagctc cctggggaga
    10861 agaagaacgg cctgtttggt aatcttatcg ccctgtcact cgggctgacc cccaacttta
    10921 aatctaactt cgacctggcc gaagatgcca agcttcaact gagcaaagac acctacgatg
    10981 atgatctcga caatctgctg gcccagatcg gcgaccagta cgcagacctt tttttggcgg
    11041 caaagaacct gtcagacgcc attctgctga gtgatattct gcgagtgaac acggagatca
    11101 ccaaagctcc gctgagcgct agtatgatca agcgctatga tgagcaccac caagacttga
    11161 ctttgctgaa ggcccttgtc agacagcaac tgcctgagaa gtacaaggaa attttcttcg
    11221 atcagtctaa aaatggctac gccggataca ttgacggcgg agcaagccag gaggaatttt
    11281 acaaatttat taagcccatc ttggaaaaaa tggacggcac cgaggagctg ctggtaaagc
    11341 ttaacagaga agatctgttg cgcaaacagc gcactttcga caatggaagc atcccccacc
    11401 agattcacct gggcgaactg cacgctatcc tcaggcggca agaggatttc tacccctttt
    11461 tgaaagataa cagggaaaag attgagaaaa tcctcacatt tcggataccc tactatgtag
    11521 gccccctcgc ccggggaaat t
//
