LOCUS       sequence-376527-       13557 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Derived from LentiCRISPR v1 but expresses EGFP instead of puromycin 
            resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13557)
  AUTHORS   McComb S, Aguade-Gorgorio J, Harder L, Marovca B, Cario G, Eckert C,
            Schrappe M, Stanulla M, von Stackelberg A, Bourquin JP, Bornhauser 
            BC
  TITLE     Activation of concurrent apoptosis and necroptosis by SMAC mimetics 
            for the treatment of refractory and relapsed ALL.
  JOURNAL   Sci Transl Med. 2016 May 18;8(339):339ra70. doi: 
            10.1126/scitranslmed.aad2986.
  PUBMED    27194728
REFERENCE   2  (bases 1 to 13557)
  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: "Sci Transl 
            Med."; date: "2016-05-18"; volume: "8(339):339ra70. doi"; pages: " 
            10.1126/scitranslmed.aad2986"
COMMENT     Sequence Label: pLentiCRISPR-EGFP
FEATURES             Location/Qualifiers
     source          1..13557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /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"
     promoter        1713..1961
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1713..1733
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1884..1903
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        3843..3918
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    3966..4083
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4152..4363
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4383..4392
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4392..4415
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4422..4442
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4467..8567
                     /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             4467..8567
                     /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             8568..8615
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8622..8678
                     /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             join(8679..8681,8682..9395)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(8679..8681,8682..9392)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(8982..9001)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9329..9350
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    9410..9998
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9463..9483)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9881..9892)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10070..10303
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10381..10502
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10418..10437)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10472..10491
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10542..10677
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10697..10716)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10698..10716)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10726..10743)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10726..10742)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10735..10757)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10884..11339
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10971..10990)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11181..11202
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11365..11469
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11470..11538,11539..12330)
                     /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(11688..11707)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12501..13089
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12990..13009
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13243..13260
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13377..13398
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13413..13418,13419..13436,13437..13443)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13451..13467
                     /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     13456..13478
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13475..13491
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13475..13491
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     13510..13530
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        13512..13530
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggaccca gagagggcct atttcccatg attccttcat
     1741 atttgcatat acgatacaag gctgttagag agataattag aattaatttg actgtaaaca
     1801 caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag
     1861 ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg
     1921 atttcttggc tttatatatc ttgtggaaag gacgaaacac cggagacggt tgtaaatgag
     1981 cacacaaaat acacatgcta aaatattata ttctatgacc tttataaaat caaccaaaat
     2041 cttcttttta ataactttag tatcaataat tagaattttt atgttccttt ttgcaaactt
     2101 ttaataaaaa tgagcaaaat aaaaaaacgc tagttttagt aactcgcgtt gttttcttca
     2161 cctttaataa tagctactcc accacttgtt cctaagcggt cagctcctgc ttcaatcatt
     2221 ttttgagcat cttcaaatgt tctaactcca ccagctgctt taactaaagc attgtcttta
     2281 acaactgact tcattagttt aacatcttca aatgttgcac ctgattttga aaatcctgtt
     2341 gatgttttaa caaattctaa tccagcttca acagctattt cacaagcttt catgatttct
     2401 tcttttgtta ataaacaatt ttccataata catttaacaa catgtgatcc agctgctttt
     2461 tttacagctt tcatgtcttc taaaactaat tcataatttt tgtcttttaa tgcaccaata
     2521 tttaatacca tatcaatttc tgttgcacca tctttaattg cttcagaaac ttcgaatgct
     2581 tttgtagctg ttgtgcatgc acctagagga aaacctacaa catttgttat tcctacattt
     2641 gtgcctttta ataattcttt acaatagctt gttcaatatg aattaacaca aactgttgca
     2701 aaatcaaatt caattgcttc atcacataat tgtttaattt cagctttcgt agcatcttgt
     2761 tttaataatg tgtgatctat atatttgttt agtttcattt tttctcctat atattcattt
     2821 ttaattttaa ttctttaata atttcgtcta ctttaacttt agcgttttga acagattcac
     2881 caacacctat aaaataaatt tttagtttag gttcagttcc acttgggcga acagcaaatc
     2941 atgacttatc ttctaaataa aattttagta agtcttgtcc tggcatatta tacattccat
     3001 cgatgtagtc ttcaacatta acaactttaa gtccagcaat ttgagttaag ggtgttgctc
     3061 tcaatgattt cattaatggt tcaattttta atttcttttc ttctggttta aaattcaagt
     3121 ttaaagtgaa agtgtaatat gcacccattt ctttaaataa atcttctaaa tagtctacta
     3181 atgttttatt ttgtttttta taaaatcaag cagcctctgc tattaatata gaagcttgta
     3241 ttccatcttt atctctagct gagtcatcaa ttacatatcc ataactttct tcataagcaa
     3301 aaacaaaatt taatccgtta tcttcttctt tagcaatttc tctacccatt catttaaatc
     3361 cagttaaagt ttttacaata ttaactccat atttttcatg agcgattcta tcacccaaat
     3421 cacttgttac aaaacttgaa tatagagccg gattttttgg aatgctattt aagcgtttta
     3481 gatttgataa ttttcaatca attaaaattg gtcctgtttg atttccatct aatcttacaa
     3541 aatgaccatc atgttttatt gccattccaa atctgtcagc atctgggtca ttcataataa
     3601 taatatctgc atcatgttta ataccatatt caagcggtat ttttcatgca ggatcaaatt
     3661 ctggatttgg atttacaaca tttttaaatg tttcatcttc aaatgcatgc tcttcaacct
     3721 caataacgtt atatcctgat tcacgtaata tttttggggt aaatttagtt cctgttccat
     3781 taactgcgct aaaaataatt tttaaatctt ttttagcttc ttgctctttt ttgtacgtct
     3841 ctgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa
     3901 gtggcaccga gtcggtgctt ttttgaattc tagatcttga gacaaatggc agtattcatc
     3961 cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac
     4021 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat
     4081 tttcgggttt attacaggga cagcagagat ccactttggc gccggctcga gtggctccgg
     4141 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
     4201 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
     4261 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
     4321 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtgtcgt gacgcgggat
     4381 ccgccaccat ggattacaaa gacgatgacg ataagatggc cccaaagaag aagcggaagg
     4441 tcggtatcca cggagtccca gcagccgaca agaagtacag catcggcctg gacatcggca
     4501 ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca
     4561 aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga gccctgctgt
     4621 tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca
     4681 ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg
     4741 tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc
     4801 acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc
     4861 ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc
     4921 tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg
     4981 acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca
     5041 accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt
     5101 ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga
     5161 agaagaatgg cctgttcgga aacctgattg ccctgagcct gggcctgacc cccaacttca
     5221 agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg
     5281 acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg
     5341 ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca
     5401 ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga
     5461 ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg
     5521 accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct
     5581 acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc
     5641 tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc
     5701 agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc
     5761 tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg
     5821 gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca
     5881 tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag agcttcatcg
     5941 agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc
     6001 tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg
     6061 gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt
     6121 tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg
     6181 agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca
     6241 cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg
     6301 aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg
     6361 aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc
     6421 ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca
     6481 agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact
     6541 tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg
     6601 tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca
     6661 ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc
     6721 ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg
     6781 gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca
     6841 gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc
     6901 tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc
     6961 tgtccgacta cgatgtggac catatcgtgc ctcagagctt tctgaaggac gactccatcg
     7021 acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg
     7081 aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta
     7141 cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata
     7201 aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac
     7261 agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag
     7321 tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt
     7381 acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg
     7441 tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact
     7501 acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta
     7561 ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg
     7621 ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg
     7681 tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga
     7741 atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca
     7801 agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg
     7861 gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca
     7921 agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca
     7981 gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg
     8041 acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa
     8101 tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg
     8161 tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg
     8221 agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga
     8281 tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg
     8341 cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt
     8401 ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc
     8461 ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca
     8521 ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacaag cgtcctgctg
     8581 ctactaagaa agctggtcaa gctaagaaaa agaaagctag cgccacgaac ttctctctgt
     8641 taaagcaagc aggcgacgtg gaagaaaacc ccggtcccgt gagcaagggc gaggagctgt
     8701 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     8761 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     8821 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     8881 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     8941 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     9001 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     9061 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     9121 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     9181 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     9241 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     9301 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     9361 ggatcactct cggcatggac gagctgtaca agtaacgcgt taagtcgaca atcaacctct
     9421 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     9481 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     9541 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     9601 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     9661 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     9721 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     9781 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     9841 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     9901 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     9961 tcagacgagt cggatctccc tttgggccgc ctccccgcgt cgactttaag accaatgact
    10021 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
    10081 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
    10141 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
    10201 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
    10261 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtacgta tagtagttca
    10321 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
    10381 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
    10441 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
    10501 tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc
    10561 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
    10621 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
    10681 ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
    10741 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
    10801 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
    10861 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
    10921 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
    10981 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
    11041 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
    11101 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
    11161 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
    11221 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
    11281 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
    11341 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
    11401 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
    11461 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
    11521 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
    11581 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
    11641 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
    11701 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
    11761 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
    11821 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
    11881 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
    11941 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
    12001 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
    12061 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
    12121 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
    12181 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
    12241 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
    12301 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
    12361 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
    12421 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
    12481 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
    12541 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
    12601 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
    12661 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
    12721 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
    12781 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
    12841 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
    12901 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
    12961 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
    13021 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
    13081 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
    13141 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
    13201 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
    13261 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
    13321 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
    13381 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
    13441 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
    13501 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaa
//
