SB-CRISPR-TAL1

LOCUS       SB-CRISPR-TAL1              12164 bp DNA     circular SYN 16-MAR-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12164)
  AUTHORS   Amane
  TITLE     Direct Submission
  JOURNAL   Exported Aug 15, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..12164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..88)
                     /direction=LEFT
                     /label=pUC ori
                     /note="/application_notes=Facilitates plasmid replication 
                     in E.coli; regulates high-copy plasmid number (500-700)."
                     /note="/description=pUC origin of replication"
                     /note="/full_name=pUC origin of replication"
                     /note="/official_designation=pUC ori"
                     /note="/uuid=e53065ee22b54256bf5eff25edb79b7e"
     CDS             complement(259..1119)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAAKLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1120..1224)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1251..1706
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     1847..1863
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1873..1891
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1913..1916
                     /label=TATA
                     /label=nonstandard type:  Miscellaneous
                     /note="/application_notes=Increases Sleeping Beauty 
                     transpositionefficiency."
                     /note="/description=TA dinucleotide base-pairs"
                     /note="/official_designation=TATA"
                     /note="/uuid=01b0e78bde8343a6bfec0117ebf44b7c"
     misc_feature    complement(1917..2143)
                     /label=IR/DR(L)
                     /label=nonstandard type:  ITR
                     /note="/application_notes=Recognized by Sleeping 
                     Beautytranspose; DNA flanked by IR/DR(L) and IR/DR(R) can 
                     be transposed by sleeping beauty transpose into  into TA 
                     dinucleotide sites."
                     /note="/description=Inverted/direct repeats of Sleeping 
                     Beautytransposon (Left)"
                     /note="/official_designation=IR/DR(L)"
                     /note="/uuid=8ce65d2787884651ae37ca3e826abaf0"
     primer_bind     complement(2144..2169)
                     /label=601-5403
     primer_bind     2211..2246
                     /label=242-Cas9-F
     promoter        2226..2466
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     2226..2246
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2397..2416
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     2471..2494
                     /label=296-TAL1-F
     primer_bind     complement(2475..2498)
                     /label=296-TAL1-R
     misc_feature    2475..2494
                     /label=TAL1-KO-1
     misc_RNA        2495..2570
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        2667..2952
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        2954..3229
                     /label=chicken beta-actin promoter
     intron          3230..3457
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      3469..3478
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3478..3543
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3550..3570
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3595..7695
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7696..7743
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     7730..7769
                     /label=281-Hygro-F
     CDS             7756..7812
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             7822..8844
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     complement(8825..8867)
                     /label=281-Hygro-R
     primer_bind     complement(8875..8892)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8881..9088
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(9083..9112)
                     /label=242-Cas9-R
     promoter        9100..9599
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     protein_bind    9603..9627
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    9628..9633
                     /label=Kozak
                     /label=nonstandard type:  Miscellaneous
                     /note="/application_notes=Facilitates translation 
                     initiation ofATG start codon downstream of the Kozak 
                     sequence."
                     /note="/description=Kozak translation initiation sequence"
                     /note="/full_name=Kozak"
                     /note="/official_designation=Kozak"
                     /note="/uuid=5f18b25a1234443494a618363f09f052"
     CDS             9634..10335
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     complement(10310..10350)
                     /label=735-178-2
     primer_bind     complement(10311..10336)
                     /label=220
     primer_bind     10336..10357
                     /label=734-178-1
     protein_bind    complement(10336..10340)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    10343..10864
                     /label=rBG pA
     polyA_site      10483..10538
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     misc_feature    10935..11162
                     /label=IR/DR(R)
                     /label=nonstandard type:  ITR
                     /note="/application_notes=Recognized by Sleeping 
                     Beautytranspose; DNA flanked by IR/DR(L) and IR/DR(R) can 
                     be transposed by sleeping beauty transpose into  into TA 
                     dinucleotide sites."
                     /note="/description=Inverted/direct repeats of Sleeping 
                     Beautytransposon (Right)"
                     /note="/official_designation=IR/DR(R)"
                     /note="/uuid=f7966ed16d574adbb44bb560d4cbf7bc"
     misc_feature    complement(11163..11166)
                     /label=TATA
                     /label=nonstandard type:  Miscellaneous
                     /note="/application_notes=Increases Sleeping Beauty 
                     transpositionefficiency."
                     /note="/description=TA dinucleotide base-pairs"
                     /note="/official_designation=TATA"
                     /note="/uuid=01b0e78bde8343a6bfec0117ebf44b7c"
     primer_bind     complement(11206..11225)
                     /label=446-jc79-1
     promoter        complement(11223..11241)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11262..11278)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    11286..11302
                     /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(11310..11340)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11355..11376
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(11664..12164)
                     /direction=LEFT
                     /label=pUC ori
                     /note="/application_notes=Facilitates plasmid replication 
                     in E.coli; regulates high-copy plasmid number (500-700)."
                     /note="/description=pUC origin of replication"
                     /note="/full_name=pUC origin of replication"
                     /note="/official_designation=pUC ori"
                     /note="/uuid=e53065ee22b54256bf5eff25edb79b7e"
ORIGIN
        1 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
       61 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
      121 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
      181 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
      241 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
      301 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
      361 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
      421 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
      481 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
      541 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
      601 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
      661 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaactt
      721 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
      781 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
      841 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
      901 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
      961 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     1021 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     1081 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     1141 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     1201 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg
     1261 tagcggcgca ttaagcgcgg cgggagtggt ggttacgcgc agcgtgaccg ctacacttgc
     1321 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     1381 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     1441 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     1501 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     1561 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     1621 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     1681 taacaaaata ttaacgctta caatttccat tcgccattca ggctgcgcaa ctgttgggaa
     1741 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     1801 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     1861 agtgagcgcg cgtaatacga ctcactatag ggcgaattgg agctcggatc cctatacagt
     1921 tgaagtcgga agtttacata cacttaagtt ggagtcatta aaactcgttt ttcaactact
     1981 ccacaaattt cttgttaaca aacaatagtt ttggcaagtc agttaggaca tctactttgt
     2041 gcatgacaca agtcattttt ccaacaattg tttacagaca gattatttca cttataattc
     2101 actgtatcac aattccagtg ggtcagaagt ttacatacac taagttgact gtgcctttaa
     2161 acagcttgga aaattccaga aaatgatgtc atggctttag aagctagatc tggccatcta
     2221 gagatgaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
     2281 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
     2341 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
     2401 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
     2461 aaggacgaaa caccgaagat acgccgcaca acttgtttta gagctagaaa tagcaagtta
     2521 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttgttt
     2581 tagagctaga aatagcaagt taaaataagg ctagtccgtt tttagcgcgt gcgccaattc
     2641 tgcagacaaa tggctctaga ggtacccgtt acataactta cggtaaatgg cccgcctggc
     2701 tgaccgccca acgacccccg cccattgacg tcaatagtaa cgccaatagg gactttccat
     2761 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     2821 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattgt
     2881 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     2941 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
     3001 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg
     3061 gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag
     3121 gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc
     3181 gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc
     3241 gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg
     3301 actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa
     3361 ttagctgagc aagaggtaag ggtttaaggg atggttggtt ggtggggtat taatgtttaa
     3421 ttacctggag cacctgcctg aaatcacttt ttttcaggtt ggaccggtgc caccatggac
     3481 tataaggacc acgacggaga ctacaaggat catgatattg attacaaaga cgatgacgat
     3541 aagatggccc caaagaagaa gcggaaggtc ggtatccacg gagtcccagc agccgacaag
     3601 aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac
     3661 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
     3721 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg
     3781 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
     3841 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
     3901 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
     3961 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
     4021 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
     4081 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
     4141 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
     4201 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa
     4261 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc
     4321 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
     4381 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
     4441 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
     4501 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
     4561 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
     4621 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt
     4681 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
     4741 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg
     4801 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
     4861 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
     4921 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
     4981 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
     5041 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
     5101 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag
     5161 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
     5221 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
     5281 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
     5341 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
     5401 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
     5461 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
     5521 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
     5581 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
     5641 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
     5701 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
     5761 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
     5821 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
     5881 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
     5941 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
     6001 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
     6061 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct
     6121 cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac
     6181 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
     6241 cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc
     6301 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
     6361 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
     6421 gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
     6481 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
     6541 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
     6601 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
     6661 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
     6721 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
     6781 gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg
     6841 cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
     6901 ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag
     6961 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
     7021 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
     7081 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
     7141 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
     7201 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga
     7261 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
     7321 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag
     7381 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
     7441 gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga
     7501 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
     7561 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
     7621 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct
     7681 cagctgggag gcgacaaaag gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa
     7741 aaggaattcg ctagcgcgac gaattttagt ctactgaaac aagcgggaga cgtggaggaa
     7801 aaccctggac ctatcgatgg tatgaaaaag cctgaactca ccgcgacgtc tgtcgagaag
     7861 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     7921 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     7981 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
     8041 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc
     8101 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
     8161 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
     8221 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
     8281 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
     8341 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
     8401 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
     8461 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc
     8521 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
     8581 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa
     8641 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
     8701 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
     8761 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc
     8821 agcactcgtc cgagggcaaa ggaataagaa ttctaactag agctcgctga tcagcctcga
     8881 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     8941 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     9001 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     9061 gggaagagaa tagcaggcat gctggggagc ggccgcatcg ggtaggggag gcgcttttcc
     9121 caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg
     9181 gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg
     9241 gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc
     9301 agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga
     9361 tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc
     9421 tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca
     9481 ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca
     9541 cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga
     9601 cctcaagttt gtacaaaaaa gcaggctgcc accatgagcg agctgattaa ggagaacatg
     9661 cacatgaagc tgtacatgga gggcaccgtg gacaaccatc acttcaagtg cacatccgag
     9721 ggcgaaggca agccctacga gggcacccag accatgagaa tcaaggtggt cgagggcggc
     9781 cctctcccct tcgccttcga catcctggct actagcttcc tctacggcag caagaccttc
     9841 atcaaccaca cccagggcat ccccgacttc ttcaagcagt ccttccctga gggcttcaca
     9901 tgggagagag tcaccacata cgaggacggg ggcgtgctga ccgctaccca ggacaccagc
     9961 ctccaggacg gctgcctcat ctacaacgtc aagatcagag gggtgaactt cacatccaac
    10021 ggccctgtga tgcagaagaa aacactcggc tgggaggcct tcaccgagac gctgtacccc
    10081 gctgacggcg gcctggaagg cagaaacgac atggccctga agctcgtggg cgggagccat
    10141 ctgatcgcaa acgccaagac cacatataga tccaagaaac ccgctaagaa cctcaagatg
    10201 cctggcgtct actatgtgga ctacagactg gaaagaatca aggaggccaa caacgagacc
    10261 tacgtcgagc agcacgaggt ggcagtggcc agatactgcg acctccctag caaactgggg
    10321 cacaagctta attgagtggt gatcctcagg tgcaggctgc ctatcagaag gtggtggctg
    10381 gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt
    10441 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
    10501 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
    10561 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
    10621 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc
    10681 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
    10741 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
    10801 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga
    10861 gatcatctgt ggaaggctac tcgaaatgtt tgacccaagt taaacaattt aaaggcaatg
    10921 ctaccaaata ctaattgagt gtatgtaaac ttctgaccca ctgggaatgt gatgaaagaa
    10981 ataaaagctg aaatgaatca ttctctctac tattattctg atatttcaca ttcttaaaat
    11041 aaagtggtga tcctaactga cctaagacag ggaattttta ctaggattaa atgtcaggaa
    11101 ttgtgaaaaa gtgagtttaa atgtatttgg ctaaggtgta tgtaaacttc cgacttcaac
    11161 tgtataggga tcctctagct agagtcgacc tcgagggggg gcccggtacc cagcttttgt
    11221 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
    11281 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
    11341 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
    11401 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
    11461 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
    11521 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
    11581 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
    11641 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
    11701 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
    11761 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
    11821 tccgcctttc tcccttcggg aaccgtggcg ctttctcata gctcacgctg taggtatctc
    11881 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
    11941 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
    12001 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
    12061 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
    12121 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtag
//