LOCUS       sequence-178320-       12817 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Advanced lentiviral CRISPR-Cas9 vector for induction of chromosomal 
            translocations; control, MLL-sgRNA + luciferase-sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12817)
  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 12817)
  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-ctrl)
FEATURES             Location/Qualifiers
     source          1..12817
                     /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"
     regulatory      4051..4060
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4060..4083
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4090..4110
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4135..8235
                     /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             4135..8235
                     /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             8236..8283
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8302..8358
                     /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      8614..8623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    9085..9673
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9138..9158)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9556..9567)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9745..9978
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10056..10177
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10093..10112)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10147..10166
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10217..10352
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10372..10391)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10373..10391)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10401..10418)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10401..10417)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10410..10432)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10559..11014
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10646..10665)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10856..10877
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11040..11144
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11145..11213,11214..12005)
                     /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(11363..11382)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12176..12764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12665..12684
                     /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 aaacaccgtc caaaacaaca acggcggcgt 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 cgagctgcag taacgccatt ttgcaaggca tggaaaaata
     3841 ccaaaccaag aatagagaag ttcagatcaa gggcgggtac atgaaaatag ctaacgttgg
     3901 gccaaacagg atatctgcgg tgagcagttt cggccccggc ccggggccaa gaacagmtgg
     3961 tgwycgcgcg cttctgcttc ccgagctcta taaaagagct cacaacccct cactcggcgc
     4021 gccagtcctc cgacagactg agtcggatcc gccaccatgg attacaaaga cgatgacgat
     4081 aagatggccc caaagaagaa gcggaaggtc ggtatccacg gagtcccagc agccgacaag
     4141 aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac
     4201 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
     4261 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg
     4321 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
     4381 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
     4441 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
     4501 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
     4561 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
     4621 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
     4681 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
     4741 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa
     4801 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc
     4861 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
     4921 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
     4981 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
     5041 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
     5101 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
     5161 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt
     5221 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
     5281 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg
     5341 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
     5401 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
     5461 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
     5521 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
     5581 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
     5641 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag
     5701 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
     5761 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
     5821 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
     5881 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
     5941 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
     6001 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
     6061 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
     6121 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
     6181 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
     6241 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
     6301 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
     6361 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
     6421 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
     6481 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
     6541 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
     6601 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct
     6661 cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac
     6721 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
     6781 cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc
     6841 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
     6901 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
     6961 gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
     7021 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
     7081 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
     7141 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
     7201 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
     7261 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
     7321 gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg
     7381 cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
     7441 ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag
     7501 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
     7561 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
     7621 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
     7681 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
     7741 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga
     7801 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
     7861 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag
     7921 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
     7981 gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga
     8041 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
     8101 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
     8161 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct
     8221 cagctgggag gcgacaagcg tcctgctgct actaagaaag ctggtcaagc taagaaaaag
     8281 aaagctagcg gaggaagcgg cgccaccaat ttcagcctgc tgaaacaggc cggcgacgtg
     8341 gaagagaacc ctggccctat ggtgtccaag ggcgaagagg acaacatggc cagcctgcct
     8401 gccacccacg agctgcacat cttcggcagc atcaacggcg tggacttcga catggtggga
     8461 cagggcaccg gcaaccccaa cgacggctac gaggaactga acctgaagtc cacaaagggc
     8521 gacctgcagt tcagcccctg gattctggtg ccccacatcg gctacggctt ccaccagtac
     8581 ctgccctacc ccgacggcat gagccctttc caagccgcca tggtggatgg cagcggctac
     8641 caggtgcacc ggaccatgca gtttgaggac ggcgccagcc tgaccgtgaa ctacaggtac
     8701 acatacgagg gcagccacat caagggcgag gcccaagtga agggcacagg ctttccagcc
     8761 gacggccccg tgatgaccaa tagcctgaca gccgccgact ggtgcagaag caagaaaacc
     8821 taccccaatg acaagaccat catcagcacc ttcaagtggt cctacaccac cggcaatggc
     8881 aagcggtaca gaagcaccgc ccggaccacc tacaccttcg ccaaacccat ggccgccaac
     8941 tacctgaaga accagcctat gtacgtgttc cgcaagaccg agctgaagca ctccaagaca
     9001 gaactgaact tcaaagagtg gcagaaagcc ttcaccgacg tgatgggcat ggacgagctg
     9061 tacaagtgaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     9121 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     9181 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     9241 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     9301 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     9361 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     9421 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     9481 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     9541 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     9601 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     9661 gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     9721 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     9781 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     9841 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     9901 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     9961 tgtggaaaat ctctagcagt acgtatagta gttcatgtca tcttattatt cagtatttat
    10021 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
    10081 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
    10141 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
    10201 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
    10261 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
    10321 ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt
    10381 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
    10441 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
    10501 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
    10561 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
    10621 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
    10681 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
    10741 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
    10801 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
    10861 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
    10921 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
    10981 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
    11041 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
    11101 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
    11161 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
    11221 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
    11281 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
    11341 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
    11401 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
    11461 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
    11521 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
    11581 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
    11641 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
    11701 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
    11761 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
    11821 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
    11881 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
    11941 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
    12001 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
    12061 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
    12121 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
    12181 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
    12241 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
    12301 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
    12361 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
    12421 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
    12481 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
    12541 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
    12601 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
    12661 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
    12721 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
    12781 cctttttacg gttcctggcc ttttgctggc cttttgc
//
