LOCUS       sequence-174984-       11472 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR interference.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11472)
  AUTHORS   Schwarzer A, Emmrich S, Schmidt F, Beck D, Ng M, Reimer C, Adams FF,
            Grasedieck S, Witte D, Kabler S, Wong JWH, Shah A, Huang Y, Jammal 
            R, Maroz A, Jongen-Lavrencic M, Schambach A, Kuchenbauer F, Pimanda 
            JE, Reinhardt D, Heckl D, Klusmann JH
  TITLE     The non-coding RNA landscape of human hematopoiesis and leukemia.
  JOURNAL   Nat Commun. 2017 Aug 9;8(1):218. doi: 10.1038/s41467-017-00212-4.
  PUBMED    28794406
REFERENCE   2  (bases 1 to 11472)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2017-08-9"; volume: "8(1):218. doi"; pages: " 
            10.1038/s41467-017-00212-4"
COMMENT     Sequence Label: pLKO5d.SFFV.dCas9-KRAB.P2A.BSD
FEATURES             Location/Qualifiers
     source          1..11472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    41..158
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        216..623
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      623..632
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             635..661
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             665..685
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(695..718,719..721,722..3277,3278..3280,3281..4795)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(695..718,719..721,722..3208,3209..3211,3212..4795)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4796..4843
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4856..4876
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4925..5110
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             5132..5188
                     /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             5189..5584
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    5618..6206
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5671..5691)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6089..6100)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6278..6511
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6589..6710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6626..6645)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6680..6699
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6750..6885
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6905..6924)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6906..6924)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6934..6951)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6934..6950)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6943..6965)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7092..7547
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7179..7198)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7389..7410
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7573..7677
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7678..7746,7747..8538)
                     /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(7896..7915)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8709..9297
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9198..9217
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9451..9468
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9585..9606
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9621..9626,9627..9644,9645..9651)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9659..9675
                     /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     9664..9686
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9683..9699
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9683..9699
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9718..9738
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9720..9738
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9766..9992
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             9993..10173
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    10220..10345
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10838..11071
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11256..11300
                     /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"
ORIGIN
        1 aattctagat cttgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
       61 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
      121 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
      181 gagatccact ttggcgccgg ctcgagcgag ctgcagtaac gccattttgc aaggcatgga
      241 aaaataccaa accaagaata gagaagttca gatcaagggc gggtacatga aaatagctaa
      301 cgttgggcca aacaggatat ctgcggtgag cagtttcggc cccggcccgg ggccaagaac
      361 agatggtcac cgcagtttcg gccccggccc gaggccaaga acagatggtc cccagatatg
      421 gcccaaccct cagcagtttc ttaagaccca tcagatgttt ccaggctccc ccaaggacct
      481 gaaatgaccc tgcgccttat ttgaattaac caatcagcct gcttctcgct tctgttcgcg
      541 cgcttctgct tcccgagctc tataaaagag ctcacaaccc ctcactcggc gcgccagtcc
      601 tccgacagac tgagtcggat ccgccaccat gggctaccca tacgatgttc cagattacgc
      661 ttcgccgaag aaaaagcgca aggtcgaagc gtccgacaag aagtacagca tcggcctggc
      721 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
      781 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
      841 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
      901 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
      961 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
     1021 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
     1081 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
     1141 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
     1201 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
     1261 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
     1321 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
     1381 cggcgagaag aagaatggcc tgttcggcaa cctgattgcc ctgagcctgg gcctgacccc
     1441 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
     1501 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
     1561 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
     1621 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
     1681 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
     1741 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
     1801 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
     1861 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
     1921 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
     1981 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
     2041 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
     2101 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
     2161 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
     2221 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
     2281 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
     2341 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
     2401 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
     2461 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
     2521 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
     2581 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
     2641 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
     2701 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
     2761 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
     2821 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
     2881 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
     2941 gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
     3001 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
     3061 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
     3121 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
     3181 caaccggctg tccgactacg atgtggacgc catcgtgcct cagagctttc tgaaggacga
     3241 ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt
     3301 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
     3361 gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
     3421 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
     3481 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
     3541 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
     3601 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
     3661 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta
     3721 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg
     3781 caaggctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
     3841 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
     3901 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
     3961 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
     4021 cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa
     4081 gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
     4141 aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga
     4201 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
     4261 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
     4321 gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc
     4381 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
     4441 ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat
     4501 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt
     4561 gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat
     4621 ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac
     4681 catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca
     4741 gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaagcg
     4801 tcctgctgct actaagaaag ctggtcaagc taagaaaaag aaagctagag gctctcccaa
     4861 gaaaaagaga aaggttgaag gcggaggcgg tggcatggat gctaagtcac taactgcctg
     4921 gtcccggaca ctggtgacct tcaaggatgt atttgtggac ttcaccaggg aggagtggaa
     4981 gctgctggac actgctcagc agatcgtgta cagaaatgtg atgctggaga actataagaa
     5041 cctggtttcc ttgggttatc agcttactaa gccagatgtg atcctccggt tggagaaggg
     5101 agaagagccc ggcggagcta gcggcagcgg cgccaccaac ttcagcctgc tgaagcaggc
     5161 cggcgacgtg gaggagaacc ccggccccat ggccaagcct ttgtctcaag aagaatccac
     5221 cctcattgaa agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt
     5281 cgccagcgca gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt
     5341 tactggggga ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg
     5401 caacctgact tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg
     5461 acggtgccga caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag
     5521 tgatggacag ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga
     5581 gggcctgcag ctgcagtagt aaacgcgtta agtcgacaat caacctctgg attacaaaat
     5641 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5701 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5761 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5821 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5881 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     5941 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     6001 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     6061 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     6121 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     6181 gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct
     6241 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     6301 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     6361 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     6421 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     6481 cccttttagt cagtgtggaa aatctctagc agtacgtata gtagttcatg tcatcttatt
     6541 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     6601 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6661 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     6721 ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     6781 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     6841 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa
     6901 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     6961 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     7021 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     7081 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     7141 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     7201 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     7261 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     7321 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     7381 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     7441 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     7501 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     7561 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     7621 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     7681 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     7741 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     7801 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     7861 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     7921 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     7981 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     8041 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     8101 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     8161 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     8221 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     8281 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     8341 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     8401 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     8461 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     8521 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     8581 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     8641 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     8701 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     8761 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     8821 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     8881 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     8941 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     9001 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     9061 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     9121 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     9181 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     9241 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     9301 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     9361 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     9421 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     9481 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     9541 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     9601 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     9661 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
     9721 attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa
     9781 tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga
     9841 aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa
     9901 ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata
     9961 ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag
    10021 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
    10081 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
    10141 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
    10201 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
    10261 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
    10321 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
    10381 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
    10441 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
    10501 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
    10561 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
    10621 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
    10681 agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat
    10741 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
    10801 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
    10861 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac
    10921 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
    10981 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
    11041 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
    11101 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt
    11161 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
    11221 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
    11281 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
    11341 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
    11401 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
    11461 ccccgagggg ac
//
