LOCUS       sequence-473226-       12474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral expression of Cas9 and encodes CT47 sgRNA for CLEC12A 
            deletion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12474)
  AUTHORS   Mohanty S, Charles Cano F, Gabdoulline R, Lai CK, Othman B, 
            Sudarsanam H, Eder T, Grebien F, Lipka DB, Henschler R, Heuser M
  TITLE     Clec12a is required for the pathogenesis of NUP98::NSD1 AML.
  JOURNAL   Blood Adv. 2025 May 7:bloodadvances.2024015739. doi: 
            10.1182/bloodadvances.2024015739.
  PUBMED    40334081
REFERENCE   2  (bases 1 to 12474)
  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: "Blood Adv. 
            2025 May 7:bloodadvances.2024015739. doi: 
            10.1182/bloodadvances.2024015739."
COMMENT     Sequence Label: sgRNA Clec12a (CT47)
FEATURES             Location/Qualifiers
     source          1..12474
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        405..616
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      636..645
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             645..668
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             675..695
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             720..4820
                     /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             720..4820
                     /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             4821..4868
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4887..4943
                     /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             4944..5645
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(5077..5095)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5345..5364
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5539..5562
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    5664..6252
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5717..5737)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6135..6146)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6324..6557
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6635..6756
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6672..6691)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6726..6745
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6796..6931
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6951..6970)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6952..6970)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6980..6997)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6980..6996)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6989..7011)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7138..7593
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7225..7244)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7435..7456
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7619..7723
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7724..7792,7793..8584)
                     /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     complement(7942..7961)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8755..9343
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9244..9263
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9497..9514
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9631..9652
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9667..9672,9673..9690,9691..9697)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9705..9721
                     /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)."
     primer_bind     9710..9732
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9729..9745
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9729..9745
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9764..9784
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9766..9784
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        9829..10208
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     enhancer        10265..10456
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(10288..10308)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10475..10701
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             10702..10882
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    10929..11054
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11547..11780
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11965..12009
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    12307..12424
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg catgtccaaa gggttcagaa gtttaagagc tatgctggaa acagcatagc
      301 aagtttaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
      361 ttgaattctc ctctagaact cgagtggctc cggtgcccgt cagtgggcag agcgcacatc
      421 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
      481 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
      541 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
      601 tgccgccaga acacaggtgt cgtgacgcgg gatccgccac catggattac aaagacgatg
      661 acgataagat ggccccaaag aagaagcgga aggtcggtat ccacggagtc ccagcagccg
      721 acaagaagta cagcatcggc ctggacatcg gcaccaactc tgtgggctgg gccgtgatca
      781 ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca
      841 gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca
      901 cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc
      961 tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg
     1021 aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca
     1081 tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac
     1141 tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga
     1201 tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg
     1261 acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca
     1321 acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc
     1381 tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggaaacctga
     1441 ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg
     1501 ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga
     1561 tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc
     1621 tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga
     1681 tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc
     1741 agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct
     1801 acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa
     1861 agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc
     1921 agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca
     1981 ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga
     2041 agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat
     2101 tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg
     2161 tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc
     2221 tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata
     2281 acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg
     2341 gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga
     2401 agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg
     2461 gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca
     2521 aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga
     2581 ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc
     2641 tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc
     2701 tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt
     2761 tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc
     2821 tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg
     2881 agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga
     2941 aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg
     3001 aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga
     3061 agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg
     3121 aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata
     3181 tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccatatcg
     3241 tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca
     3301 agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact
     3361 actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca
     3421 aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg
     3481 tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta
     3541 agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc
     3601 tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc
     3661 accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc
     3721 ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga
     3781 tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca
     3841 tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc
     3901 tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca
     3961 ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga
     4021 caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca
     4081 gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt
     4141 ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag
     4201 agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc
     4261 tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact
     4321 ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga
     4381 agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact
     4441 atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc
     4501 acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc
     4561 tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca
     4621 tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg
     4681 ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg
     4741 tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc
     4801 tgtctcagct gggaggcgac aagcgtcctg ctgctactaa gaaagctggt caagctaaga
     4861 aaaagaaagc tagcggagga agcggcgcca ccaatttcag cctgctgaaa caggccggcg
     4921 acgtggaaga gaaccctggc cctatggtga gcaagggcga ggaggtcatc aaagagttca
     4981 tgcgcttcaa ggtgcgcatg gagggctcca tgaacggcca cgagttcgag atcgagggcg
     5041 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg
     5101 gccccctgcc cttcgcctgg gacatcctgt ccccccagtt catgtacggc tccaaggcgt
     5161 acgtgaagca ccccgccgac atccccgatt acaagaagct gtccttcccc gagggcttca
     5221 agtgggagcg cgtgatgaac ttcgaggacg gcggtctggt gaccgtgacc caggactcct
     5281 ccctgcagga cggcacgctg atctacaagg tgaagatgcg cggcaccaac ttcccccccg
     5341 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag cgcctgtacc
     5401 cccgcgacgg cgtgctgaag ggcgagatcc accaggccct gaagctgaag gacggcggcc
     5461 actacctggt ggagttcaag accatctaca tggccaagaa gcccgtgcaa ctgcccggct
     5521 actactacgt ggacaccaag ctggacatca cctcccacaa cgaggactac accatcgtgg
     5581 aacagtacga gcgctccgag ggccgccacc acctgttcct gtacggcatg gacgagctgt
     5641 acaagtaaac gcgttaagtc gacaatcaac ctctggatta caaaatttgt gaaagattga
     5701 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     5761 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     5821 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     5881 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     5941 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     6001 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     6061 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     6121 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     6181 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     6241 ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag atcttagcca
     6301 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct
     6361 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     6421 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     6481 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     6541 gtggaaaatc tctagcagta cgtatagtag ttcatgtcat cttattattc agtatttata
     6601 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
     6661 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6721 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
     6781 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     6841 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     6901 cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg
     6961 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     7021 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     7081 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
     7141 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     7201 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     7261 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     7321 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     7381 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     7441 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     7501 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     7561 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     7621 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     7681 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     7741 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     7801 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     7861 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     7921 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     7981 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     8041 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     8101 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     8161 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     8221 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     8281 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     8341 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     8401 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     8461 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     8521 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     8581 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     8641 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     8701 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     8761 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     8821 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     8881 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     8941 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     9001 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     9061 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     9121 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     9181 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     9241 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     9301 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     9361 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     9421 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     9481 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     9541 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     9601 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     9661 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     9721 atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta
     9781 aagggaacaa aagctggagc tgcaagcggt accgtcgacc tgcaggttga cattgattat
     9841 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     9901 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     9961 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
    10021 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
    10081 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
    10141 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
    10201 ttaccatggc gcgccatgca gggcctgaaa taacctctga aagaggaact tggttaggta
    10261 ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
    10321 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
    10381 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
    10441 cagcaaccat agtcccttaa gaggcgcgcc ttaatgtagt cttatgcaat actcttgtag
    10501 tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt
    10561 gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg
    10621 ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag
    10681 tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc
    10741 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
    10801 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
    10861 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
    10921 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
    10981 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
    11041 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
    11101 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
    11161 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
    11221 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
    11281 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
    11341 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg
    11401 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
    11461 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
    11521 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
    11581 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
    11641 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
    11701 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
    11761 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
    11821 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
    11881 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
    11941 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
    12001 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcata
    12061 atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat
    12121 agagttaggc agggatattc accattatcg tttcagaccc acctcccaac cccgagggga
    12181 cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt
    12241 cgattagtga acggatcggc actgcgtgcg ccaattctgc agacaaatgg cagtattcat
    12301 ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
    12361 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
    12421 ttttcgggtt tattacaggg acagcagaga tccagtttgg ttaattaagg tacc
//
