LOCUS       sequence-392598-       12346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  All-in-vector of SpCas9 gRNA cloning sites (BsmBI) and 
            CMVp-Target-AID expression unit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12346)
  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 12346)
  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: pSI-218
FEATURES             Location/Qualifiers
     source          1..12346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(56..511)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(193..214)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     405..424
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     638..660
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     652..669
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     653..669
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        679..697
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        764..1012
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     764..784
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     935..954
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2894..2969
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        2983..3362
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3363..3566
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3517..3537
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3611..3629
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3658..3678
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3703..3729,3730..3732,3733..7806)
                     /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             3703..7806
                     /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"
     CDS             7819..7839
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8047..8112
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             8746..8766
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8773..9024
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             9025..9078
                     /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             9085..9879
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(9139..9158)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9749..9768
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    10053..10174
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10090..10109)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10144..10163
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10211..10231)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(10211..10229)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10250..10266)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10250..10266)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10263..10285)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10274..10290
                     /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(10298..10304,10305..10322,10323..10328))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10343..10364
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10481..10498)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10652..11240)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10732..10751)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11411..12202,12203..12271))
                     /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     12034..12053
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(12272..12346,1..30))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
       61 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
      121 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
      181 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
      241 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
      301 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
      361 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
      421 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
      481 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
      541 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      601 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      661 acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac cgggccaatt
      721 cgaacgctga cgtcatcaac ccgctccaag gaatcgcggg cccgagggcc tatttcccat
      781 gattccttca tatttgcata tacgatacaa ggctgttaga gagataatta gaattaattt
      841 gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg
      901 gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg
      961 aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccggagacgg
     1021 ttgtaaatga gcacacaaaa tacacatgct aaaatattat attctatgac ctttataaaa
     1081 tcaaccaaaa tcttcttttt aataacttta gtatcaataa ttagaatttt tatgttcctt
     1141 tttgcaaact tttaataaaa atgagcaaaa taaaaaaacg ctagttttag taactcgcgt
     1201 tgttttcttc acctttaata atagctactc caccacttgt tcctaagcgg tcagctcctg
     1261 cttcaatcat tttttgagca tcttcaaatg ttctaactcc accagctgct ttaactaaag
     1321 cattgtcttt aacaactgac ttcattagtt taacatcttc aaatgttgca cctgattttg
     1381 aaaatcctgt tgatgtttta acaaattcta atccagcttc aacagctatt tcacaagctt
     1441 tcatgatttc ttcttttgtt aataaacaat tttccataat acatttaaca acatgtgatc
     1501 cagctgcttt ttttacagct ttcatgtctt ctaaaactaa ttcataattt ttgtctttta
     1561 atgcaccaat atttaatacc atatcaattt ctgttgcacc atctttaatt gcttcagaaa
     1621 cttcgaatgc ttttgtagct gttgtgcatg cacctagagg aaaacctaca acatttgtta
     1681 ttcctacatt tgtgcctttt aataattctt tacaatagct tgttcaatat gaattaacac
     1741 aaactgttgc aaaatcaaat tcaattgctt catcacataa ttgtttaatt tcagctttcg
     1801 tagcatcttg ttttaataat gtgtgatcta tatatttgtt tagtttcatt ttttctccta
     1861 tatattcatt tttaatttta attctttaat aatttcgtct actttaactt tagcgttttg
     1921 aacagattca ccaacaccta taaaataaat ttttagttta ggttcagttc cacttgggcg
     1981 aacagcaaat catgacttat cttctaaata aaattttagt aagtcttgtc ctggcatatt
     2041 atacattcca tcgatgtagt cttcaacatt aacaacttta agtccagcaa tttgagttaa
     2101 gggtgttgct ctcaatgatt tcattaatgg ttcaattttt aatttctttt cttctggttt
     2161 aaaattcaag tttaaagtga aagtgtaata tgcacccatt tctttaaata aatcttctaa
     2221 atagtctact aatgttttat tttgtttttt ataaaatcaa gcagcctctg ctattaatat
     2281 agaagcttgt attccatctt tatctctagc tgagtcatca attacatatc cataactttc
     2341 ttcataagca aaaacaaaat ttaatccgtt atcttcttct ttagcaattt ctctacccat
     2401 tcatttaaat ccagttaaag tttttacaat attaactcca tatttttcat gagcgattct
     2461 atcacccaaa tcacttgtta caaaacttga atatagagcc ggattttttg gaatgctatt
     2521 taagcgtttt agatttgata attttcaatc aattaaaatt ggtcctgttt gatttccatc
     2581 taatcttaca aaatgaccat catgttttat tgccattcca aatctgtcag catctgggtc
     2641 attcataata ataatatctg catcatgttt aataccatat tcaagcggta tttttcatgc
     2701 aggatcaaat tctggatttg gatttacaac atttttaaat gtttcatctt caaatgcatg
     2761 ctcttcaacc tcaataacgt tatatcctga ttcacgtaat atttttgggg taaatttagt
     2821 tcctgttcca ttaactgcgc taaaaataat ttttaaatct tttttagctt cttgctcttt
     2881 tttgtacgtc tctgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
     2941 aacttgaaaa agtggcaccg agtcggtgct tttttacgcg ttgacattga ttattgacta
     3001 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     3061 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     3121 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     3181 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     3241 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     3301 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     3361 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     3421 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     3481 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     3541 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
     3601 ttatcgaaat taatacgact cactataggg agacccaagc tggctagcga aatggcaccg
     3661 aagaagaagc gtaaagtcgg aatccacgga gttcctgcgg caatggacaa gaagtactcc
     3721 attgggctcg ctatcggcac aaacagcgtc ggttgggccg tcattacgga cgagtacaag
     3781 gtgccgagca aaaaattcaa agttctgggc aataccgatc gccacagcat aaagaagaac
     3841 ctcattggcg ccctcctgtt cgactccggg gaaacggccg aagccacgcg gctcaaaaga
     3901 acagcacggc gcagatatac ccgcagaaag aatcggatct gctacctgca ggagatcttt
     3961 agtaatgaga tggctaaggt ggatgactct ttcttccata ggctggagga gtcctttttg
     4021 gtggaggagg ataaaaagca cgagcgccac ccaatctttg gcaatatcgt ggacgaggtg
     4081 gcgtaccatg aaaagtaccc aaccatatat catctgagga agaagcttgt agacagtact
     4141 gataaggctg acttgcggtt gatctatctc gcgctggcgc atatgatcaa atttcgggga
     4201 cacttcctca tcgaggggga cctgaaccca gacaacagcg atgtcgacaa actctttatc
     4261 caactggttc agacttacaa tcagcttttc gaagagaacc cgatcaacgc atccggagtt
     4321 gacgccaaag caatcctgag cgctaggctg tccaaatccc ggcggctcga aaacctcatc
     4381 gcacagctcc ctggggagaa gaagaacggc ctgtttggta atcttatcgc cctgtcactc
     4441 gggctgaccc ccaactttaa atctaacttc gacctggccg aagatgccaa gcttcaactg
     4501 agcaaagaca cctacgatga tgatctcgac aatctgctgg cccagatcgg cgaccagtac
     4561 gcagaccttt ttttggcggc aaagaacctg tcagacgcca ttctgctgag tgatattctg
     4621 cgagtgaaca cggagatcac caaagctccg ctgagcgcta gtatgatcaa gcgctatgat
     4681 gagcaccacc aagacttgac tttgctgaag gcccttgtca gacagcaact gcctgagaag
     4741 tacaaggaaa ttttcttcga tcagtctaaa aatggctacg ccggatacat tgacggcgga
     4801 gcaagccagg aggaatttta caaatttatt aagcccatct tggaaaaaat ggacggcacc
     4861 gaggagctgc tggtaaagct taacagagaa gatctgttgc gcaaacagcg cactttcgac
     4921 aatggaagca tcccccacca gattcacctg ggcgaactgc acgctatcct caggcggcaa
     4981 gaggatttct accccttttt gaaagataac agggaaaaga ttgagaaaat cctcacattt
     5041 cggataccct actatgtagg ccccctcgcc cggggaaatt ccagattcgc gtggatgact
     5101 cgcaaatcag aagagaccat cactccctgg aacttcgagg aagtcgtgga taagggggcc
     5161 tctgcccagt ccttcatcga aaggatgact aactttgata aaaatctgcc taacgaaaag
     5221 gtgcttccta aacactctct gctgtacgag tacttcacag tttataacga gctcaccaag
     5281 gtcaaatacg tcacagaagg gatgagaaag ccagcattcc tgtctggaga gcagaagaaa
     5341 gctatcgtgg acctcctctt caagacgaac cggaaagtta ccgtgaaaca gctcaaagaa
     5401 gactatttca aaaagattga atgtttcgac tctgttgaaa tcagcggagt ggaggatcgc
     5461 ttcaacgcat ccctgggaac gtatcacgat ctcctgaaaa tcattaaaga caaggacttc
     5521 ctggacaatg aggagaacga ggacattctt gaggacattg tcctcaccct tacgttgttt
     5581 gaagataggg agatgattga agaacgcttg aaaacttacg ctcatctctt cgacgacaaa
     5641 gtcatgaaac agctcaagag gcgccgatat acaggatggg ggcggctgtc aagaaaactg
     5701 atcaatggga tccgagacaa gcagagtgga aagacaatcc tggattttct taagtccgat
     5761 ggatttgcca accggaactt catgcagttg atccatgatg actctctcac ctttaaggag
     5821 gacatccaga aagcacaagt ttctggccag ggggacagtc ttcacgagca catcgctaat
     5881 cttgcaggta gcccagctat caaaaaggga atactgcaga ccgttaaggt cgtggatgaa
     5941 ctcgtcaaag taatgggaag gcataagccc gagaatatcg ttatcgagat ggcccgagag
     6001 aaccaaacta cccagaaggg acagaagaac agtagggaaa ggatgaagag gattgaagag
     6061 ggtataaaag aactggggtc ccaaatcctt aaggaacacc cagttgaaaa cacccagctt
     6121 cagaatgaga agctctacct gtactacctg cagaacggca gggacatgta cgtggatcag
     6181 gaactggaca tcaatcggct ctccgactac gacgtggatc atatcgtgcc ccagtctttt
     6241 ctcaaagatg attctattga taataaagtg ttgacaagat ccgataaaaa tagagggaag
     6301 agtgataacg tcccctcaga agaagttgtc aagaaaatga aaaattattg gcggcagctg
     6361 ctgaacgcca aactgatcac acaacggaag ttcgataatc tgactaaggc tgaacgaggt
     6421 ggcctgtctg agttggataa agccggcttc atcaaaaggc agcttgttga gacacgccag
     6481 atcaccaagc acgtggccca aattctcgat tcacgcatga acaccaagta cgatgaaaat
     6541 gacaaactga ttcgagaggt gaaagttatt actctgaagt ctaagctggt ctcagatttc
     6601 agaaaggact ttcagtttta taaggtgaga gagatcaaca attaccacca tgcgcatgat
     6661 gcctacctga atgcagtggt aggcactgca cttatcaaaa aatatcccaa gcttgaatct
     6721 gaatttgttt acggagacta taaagtgtac gatgttagga aaatgatcgc aaagtctgag
     6781 caggaaatag gcaaggccac cgctaagtac ttcttttaca gcaatattat gaattttttc
     6841 aagaccgaga ttacactggc caatggagag attcggaagc gaccacttat cgaaacaaac
     6901 ggagaaacag gagaaatcgt gtgggacaag ggtagggatt tcgcgacagt ccggaaggtc
     6961 ctgtccatgc cgcaggtgaa catcgttaaa aagaccgaag tacagaccgg aggcttctcc
     7021 aaggaaagta tcctcccgaa aaggaacagc gacaagctga tcgcacgcaa aaaagattgg
     7081 gaccccaaga aatacggcgg attcgattct cctacagtcg cttacagtgt actggttgtg
     7141 gccaaagtgg agaaagggaa gtctaaaaaa ctcaaaagcg tcaaggaact gctgggcatc
     7201 acaatcatgg agcgatcaag cttcgaaaaa aaccccatcg actttctcga ggcgaaagga
     7261 tataaagagg tcaaaaaaga cctcatcatt aagcttccca agtactctct ctttgagctt
     7321 gaaaacggcc ggaaacgaat gctcgctagt gcgggcgagc tgcagaaagg taacgagctg
     7381 gcactgccct ctaaatacgt taatttcttg tatctggcca gccactatga aaagctcaaa
     7441 gggtctcccg aagataatga gcagaagcag ctgttcgtgg aacaacacaa acactacctt
     7501 gatgagatca tcgagcaaat aagcgaattc tccaaaagag tgatcctcgc cgacgctaac
     7561 ctcgataagg tgctttctgc ttacaataag cacagggata agcccatcag ggagcaggca
     7621 gaaaacatta tccacttgtt tactctgacc aacttgggcg cgcctgcagc cttcaagtac
     7681 ttcgacacca ccatagacag aaagcggtac acctctacaa aggaggtcct ggacgccaca
     7741 ctgattcatc agtcaattac ggggctctat gaaacaagaa tcgacctctc tcagctcggt
     7801 ggagacagca gggctgaccc caagaagaag aggaaggtgg gtggaggagg taccggcggt
     7861 ggaggctcag cagaatacgt acgagctctg tttgacttca atgggaatga cgaggaggat
     7921 ctccccttta agaagggcga tattctccgc atcagagata agcccgaaga acaatggtgg
     7981 aatgccgagg atagcgaagg gaaaaggggc atgattctgg tgccatatgt ggagaaatat
     8041 tccggtgact acaaagacca tgatggggat tacaaagacc acgacatcga ctacaaagac
     8101 gacgacgata aatcagggat gacagacgcc gagtacgtgc gcattcatga gaaactggat
     8161 atttacacct tcaagaagca gttcttcaac aacaagaaat ctgtgtcaca ccgctgctac
     8221 gtgctgtttg agttgaagcg aaggggcgaa agaagggctt gcttttgggg ctatgccgtc
     8281 aacaagcccc aaagtggcac cgagagagga atacacgctg agatattcag tatccgaaag
     8341 gtggaagagt atcttcggga taatcctggg cagtttacga tcaactggta ttccagctgg
     8401 agtccttgcg ctgattgtgc cgagaaaatt ctggaatggt ataatcagga acttcgggga
     8461 aacgggcaca cattgaaaat ctgggcctgc aagctgtact acgagaagaa tgcccggaac
     8521 cagataggac tctggaatct gagggacaat ggtgtaggcc tgaacgtgat ggtttccgag
     8581 cactatcagt gttgtcggaa gattttcatc caaagctctc ataaccagct caatgaaaac
     8641 cgctggttgg agaaaacact gaaacgtgcg gagaagtgga gatccgagct gagcatcatg
     8701 atccaggtca agattctgca taccactaag tctccagccg ttggtcccaa gaagaaaaga
     8761 aaagtcggta ccatgaccaa cctttccgac atcatagaga aggaaacagg caaacagttg
     8821 gtcatccaag agtcgatact catgcttcct gaagaagttg aggaggtcat tgggaataag
     8881 ccggaaagtg acattctcgt acacactgcg tatgatgaga gcaccgatga gaacgtgatg
     8941 ctgctcacgt cagatgcccc agagtacaaa ccctgggctc tggtgattca ggactctaat
     9001 ggagagaaca agatcaagat gctagaaggc aggggaagcc ttctgacttg tggggatgtg
     9061 gaagaaaacc ctggtccatc tagaatgatt gaacaagatg gattgcacgc aggttctccg
     9121 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     9181 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     9241 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     9301 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     9361 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     9421 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     9481 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     9541 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     9601 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     9661 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     9721 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     9781 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     9841 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     9901 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     9961 atgaaaggtt gggcttcaga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
    10021 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
    10081 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
    10141 gttgtggttt gtccaaactc atcaatgtat cttagcggcc gccaccgcgg tggagctcca
    10201 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
    10261 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
    10321 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
    10381 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
    10441 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
    10501 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
    10561 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
    10621 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
    10681 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
    10741 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
    10801 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
    10861 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
    10921 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
    10981 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
    11041 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
    11101 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
    11161 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
    11221 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
    11281 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
    11341 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
    11401 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
    11461 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
    11521 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
    11581 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
    11641 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
    11701 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
    11761 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
    11821 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
    11881 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
    11941 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
    12001 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
    12061 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
    12121 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
    12181 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
    12241 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
    12301 tttatcaggg ttattgtctc atgagcggat acatatttga atgtat
//
