LOCUS       sequence-178333-       12818 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral CRISPR-Cas9 vector for induction of chromosomal 
            translocations; MLL-ENL,(t[11;19]).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12818)
  AUTHORS   Reimer J, Knoess S, Labuhn M, Charpentier EM, Gohring G, 
            Schlegelberger B, Klusmann JH, Heckl D
  TITLE     CRISPR-Cas9-induced t(11;19)/MLL-ENL translocations initiate 
            leukemia in human hematopoietic progenitor cells in vivo.
  JOURNAL   Haematologica. 2017 Jun 1. pii: haematol.2017.164046. doi: 
            10.3324/haematol.2017.164046.
  PUBMED    28572162
REFERENCE   2  (bases 1 to 12818)
  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: 
            "Haematologica. 2017 Jun 1. pii: haematol.2017.164046. doi: 
            10.3324/haematol.2017.164046."
COMMENT     Sequence Label: L-CRISPR-CTN (MLL-ENL)
FEATURES             Location/Qualifiers
     source          1..12818
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     101..118
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    235..256
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(271..276,277..294,295..301)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    309..325
                     /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     314..336
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     333..349
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     333..349
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     368..388
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        370..388
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        433..812
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     enhancer        869..1060
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(892..912)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1079..1305
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1306..1486
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1533..1658
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2151..2384
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2569..2613
                     /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    2911..3028
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3079..3327
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3079..3099
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3250..3269
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        3452..3666
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     3587..3606
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     promoter        3821..4032
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4052..4061
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4061..4084
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4091..4111
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4136..8236
                     /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             4136..8236
                     /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             8237..8284
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8303..8359
                     /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"
     regulatory      8615..8624
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    9086..9674
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9139..9159)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9557..9568)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9746..9979
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10057..10178
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10094..10113)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10148..10167
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10218..10353
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10373..10392)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10374..10392)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10402..10419)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10402..10418)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10411..10433)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10560..11015
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10647..10666)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10857..10878
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11041..11145
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11146..11214,11215..12006)
                     /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(11364..11383)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12177..12765
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12666..12685
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
       61 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      121 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
      181 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
      241 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      301 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
      361 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag cggtaccgtc
      421 gacctgcagg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
      481 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      541 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      601 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      661 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      721 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      781 acatctacgt attagtcatc gctattacca tggcgcgcca tgcagggcct gaaataacct
      841 ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt
      901 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
      961 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     1021 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc ttaagaggcg cgccttaatg
     1081 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
     1141 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
     1201 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
     1261 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
     1321 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     1381 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     1441 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     1501 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     1561 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     1621 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     1681 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     1741 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     1801 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     1861 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     1921 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     1981 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     2041 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     2101 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     2161 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     2221 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     2281 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     2341 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     2401 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     2461 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     2521 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     2581 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     2641 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     2701 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     2761 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
     2821 gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt
     2881 ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta
     2941 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     3001 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt
     3061 ttggttaatt aaggtaccga gggcctattt cccatgattc cttcatattt gcatatacga
     3121 tacaaggctg ttagagagat aattagaatt aatttgactg taaacacaaa gatattagta
     3181 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
     3241 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta
     3301 tatatcttgt ggaaaggacg aaacaccgac acgacatgtg ctcagcgggt ttaagagcta
     3361 tgctggaaac agcatagcaa gtttaaataa ggctagtccg ttatcaactt gaaaaagtgg
     3421 caccgagtcg gtgctttttt gaattcaatt cgaacgctga cgtcatcaac ccgctccaag
     3481 gaatcgcggg cccagtgtca ctaggcggga acacccagcg cgcgtgcgcc ctggcaggaa
     3541 gatggctgtg agggacaggg gagtggcgcc ctgcaatatt tgcatgtcgc tatgtgttct
     3601 gggaaatcac cataaacgtg aaatgtcttt ggatttggga atcttataag ttctgtatga
     3661 gaccacagat ctaattagca ggtgggttta gcgcgtttaa gagctatgct ggaaacagca
     3721 tagcaagttt aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
     3781 ttttttctag aaacctcgag tggctccggt gcccgtcagt gggcagagcg cacatcgccc
     3841 acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg
     3901 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
     3961 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
     4021 gccagaacac aggtgtcgtg acgcgggatc cgccaccatg gattacaaag acgatgacga
     4081 taagatggcc ccaaagaaga agcggaaggt cggtatccac ggagtcccag cagccgacaa
     4141 gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga
     4201 cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat
     4261 caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg
     4321 gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca
     4381 agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga
     4441 gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt
     4501 ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt
     4561 ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa
     4621 gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa
     4681 gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc
     4741 cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga
     4801 aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc
     4861 cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa
     4921 actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg
     4981 cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag
     5041 cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa
     5101 gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct
     5161 gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat
     5221 tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat
     5281 ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg
     5341 gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct
     5401 gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat
     5461 cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc
     5521 ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga
     5581 caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc
     5641 caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga
     5701 gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga
     5761 gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca
     5821 gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt
     5881 ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga
     5941 caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct
     6001 gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt
     6061 cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag
     6121 ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct
     6181 gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac
     6241 ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca
     6301 cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt
     6361 ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat
     6421 ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg
     6481 gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa
     6541 cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta
     6601 cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc
     6661 tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa
     6721 ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg
     6781 gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc
     6841 cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga
     6901 aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta
     6961 cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt
     7021 gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca
     7081 cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa
     7141 gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc
     7201 caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat
     7261 gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat
     7321 cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt
     7381 gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg
     7441 cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa
     7501 gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt
     7561 gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct
     7621 gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga
     7681 agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct
     7741 gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg
     7801 aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga
     7861 gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa
     7921 gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc
     7981 cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag
     8041 agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc
     8101 cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct
     8161 ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc
     8221 tcagctggga ggcgacaagc gtcctgctgc tactaagaaa gctggtcaag ctaagaaaaa
     8281 gaaagctagc ggaggaagcg gcgccaccaa tttcagcctg ctgaaacagg ccggcgacgt
     8341 ggaagagaac cctggcccta tggtgtccaa gggcgaagag gacaacatgg ccagcctgcc
     8401 tgccacccac gagctgcaca tcttcggcag catcaacggc gtggacttcg acatggtggg
     8461 acagggcacc ggcaacccca acgacggcta cgaggaactg aacctgaagt ccacaaaggg
     8521 cgacctgcag ttcagcccct ggattctggt gccccacatc ggctacggct tccaccagta
     8581 cctgccctac cccgacggca tgagcccttt ccaagccgcc atggtggatg gcagcggcta
     8641 ccaggtgcac cggaccatgc agtttgagga cggcgccagc ctgaccgtga actacaggta
     8701 cacatacgag ggcagccaca tcaagggcga ggcccaagtg aagggcacag gctttccagc
     8761 cgacggcccc gtgatgacca atagcctgac agccgccgac tggtgcagaa gcaagaaaac
     8821 ctaccccaat gacaagacca tcatcagcac cttcaagtgg tcctacacca ccggcaatgg
     8881 caagcggtac agaagcaccg cccggaccac ctacaccttc gccaaaccca tggccgccaa
     8941 ctacctgaag aaccagccta tgtacgtgtt ccgcaagacc gagctgaagc actccaagac
     9001 agaactgaac ttcaaagagt ggcagaaagc cttcaccgac gtgatgggca tggacgagct
     9061 gtacaagtga acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     9121 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     9181 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     9241 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     9301 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     9361 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     9421 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     9481 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     9541 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     9601 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     9661 ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc
     9721 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     9781 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     9841 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     9901 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     9961 gtgtggaaaa tctctagcag tacgtatagt agttcatgtc atcttattat tcagtattta
    10021 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
    10081 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
    10141 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
    10201 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
    10261 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
    10321 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag
    10381 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
    10441 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
    10501 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
    10561 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
    10621 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
    10681 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
    10741 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
    10801 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
    10861 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
    10921 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
    10981 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
    11041 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
    11101 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
    11161 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
    11221 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
    11281 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
    11341 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
    11401 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
    11461 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
    11521 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
    11581 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
    11641 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
    11701 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
    11761 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
    11821 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
    11881 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
    11941 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
    12001 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
    12061 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
    12121 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
    12181 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
    12241 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
    12301 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
    12361 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
    12421 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
    12481 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
    12541 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
    12601 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
    12661 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
    12721 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
    12781 gcctttttac ggttcctggc cttttgctgg ccttttgc
//
