LOCUS       sequence-456693-       13548 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Derived from LentiCRISPR v1 but expresses mCherry instead of 
            puromycin resistance. 3rd generation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13548)
  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 13548)
  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-mCherry
FEATURES             Location/Qualifiers
     source          1..13548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        47..273
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             274..454
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    501..626
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1119..1352
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1537..1581
                     /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        1759..2007
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1759..1779
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1930..1949
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        3889..3964
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    4012..4129
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4198..4409
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4429..4438
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4438..4461
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4468..4488
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4513..8613
                     /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             4513..8613
                     /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             8614..8661
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8668..8724
                     /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             8725..9432
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(8867..8885)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     9135..9154
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    9447..10035
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9500..9520)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9918..9929)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10107..10340
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10418..10539
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10455..10474)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10509..10528
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10579..10714
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10734..10753)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10735..10753)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10763..10780)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10763..10779)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10772..10794)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10921..11376
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11008..11027)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11218..11239
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11402..11506
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11507..11575,11576..12367)
                     /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(11725..11744)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12538..13126
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     13027..13046
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13280..13297
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13414..13435
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13450..13455,13456..13473,13474..13480)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13488..13504
                     /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     13493..13515
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13512..13528
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13512..13528
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     join(13547..13548,1..19)
                     /label=T3
                     /note="T3 promoter, forward primer"
ORIGIN
        1 aattaaccct cactaaaggg aacaaaagct ggagctgcaa gcttaatgta gtcttatgca
       61 atactcttgt agtcttgcaa catggtaacg atgagttagc aacatgcctt acaaggagag
      121 aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc cttattagga
      181 aggcaacaga cgggtctgac atggattgga cgaaccactg aattgccgca ttgcagagat
      241 attgtattta agtgcctagc tcgatacata aacgggtctc tctggttaga ccagatctga
      301 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      361 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      421 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      481 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      541 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      601 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      661 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      721 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      781 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
      841 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
      901 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
      961 aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
     1021 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
     1081 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
     1141 ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga
     1201 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
     1261 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
     1321 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
     1381 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
     1441 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
     1501 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
     1561 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
     1621 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
     1681 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
     1741 accccgaggg gacccagaga gggcctattt cccatgattc cttcatattt gcatatacga
     1801 tacaaggctg ttagagagat aattagaatt aatttgactg taaacacaaa gatattagta
     1861 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
     1921 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta
     1981 tatatcttgt ggaaaggacg aaacaccgga gacggttgta aatgagcaca caaaatacac
     2041 atgctaaaat attatattct atgaccttta taaaatcaac caaaatcttc tttttaataa
     2101 ctttagtatc aataattaga atttttatgt tcctttttgc aaacttttaa taaaaatgag
     2161 caaaataaaa aaacgctagt tttagtaact cgcgttgttt tcttcacctt taataatagc
     2221 tactccacca cttgttccta agcggtcagc tcctgcttca atcatttttt gagcatcttc
     2281 aaatgttcta actccaccag ctgctttaac taaagcattg tctttaacaa ctgacttcat
     2341 tagtttaaca tcttcaaatg ttgcacctga ttttgaaaat cctgttgatg ttttaacaaa
     2401 ttctaatcca gcttcaacag ctatttcaca agctttcatg atttcttctt ttgttaataa
     2461 acaattttcc ataatacatt taacaacatg tgatccagct gcttttttta cagctttcat
     2521 gtcttctaaa actaattcat aatttttgtc ttttaatgca ccaatattta ataccatatc
     2581 aatttctgtt gcaccatctt taattgcttc agaaacttcg aatgcttttg tagctgttgt
     2641 gcatgcacct agaggaaaac ctacaacatt tgttattcct acatttgtgc cttttaataa
     2701 ttctttacaa tagcttgttc aatatgaatt aacacaaact gttgcaaaat caaattcaat
     2761 tgcttcatca cataattgtt taatttcagc tttcgtagca tcttgtttta ataatgtgtg
     2821 atctatatat ttgtttagtt tcattttttc tcctatatat tcatttttaa ttttaattct
     2881 ttaataattt cgtctacttt aactttagcg ttttgaacag attcaccaac acctataaaa
     2941 taaattttta gtttaggttc agttccactt gggcgaacag caaatcatga cttatcttct
     3001 aaataaaatt ttagtaagtc ttgtcctggc atattataca ttccatcgat gtagtcttca
     3061 acattaacaa ctttaagtcc agcaatttga gttaagggtg ttgctctcaa tgatttcatt
     3121 aatggttcaa tttttaattt cttttcttct ggtttaaaat tcaagtttaa agtgaaagtg
     3181 taatatgcac ccatttcttt aaataaatct tctaaatagt ctactaatgt tttattttgt
     3241 tttttataaa atcaagcagc ctctgctatt aatatagaag cttgtattcc atctttatct
     3301 ctagctgagt catcaattac atatccataa ctttcttcat aagcaaaaac aaaatttaat
     3361 ccgttatctt cttctttagc aatttctcta cccattcatt taaatccagt taaagttttt
     3421 acaatattaa ctccatattt ttcatgagcg attctatcac ccaaatcact tgttacaaaa
     3481 cttgaatata gagccggatt ttttggaatg ctatttaagc gttttagatt tgataatttt
     3541 caatcaatta aaattggtcc tgtttgattt ccatctaatc ttacaaaatg accatcatgt
     3601 tttattgcca ttccaaatct gtcagcatct gggtcattca taataataat atctgcatca
     3661 tgtttaatac catattcaag cggtattttt catgcaggat caaattctgg atttggattt
     3721 acaacatttt taaatgtttc atcttcaaat gcatgctctt caacctcaat aacgttatat
     3781 cctgattcac gtaatatttt tggggtaaat ttagttcctg ttccattaac tgcgctaaaa
     3841 ataattttta aatctttttt agcttcttgc tcttttttgt acgtctctgt tttagagcta
     3901 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     3961 gtgctttttt gaattctaga tcttgagaca aatggcagta ttcatccaca attttaaaag
     4021 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     4081 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
     4141 cagggacagc agagatccac tttggcgccg gctcgagtgg ctccggtgcc cgtcagtggg
     4201 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg
     4261 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     4321 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     4381 ttcgcaacgg gtttgccgcc agaacacagg tgtcgtgacg cgggatccgc caccatggat
     4441 tacaaagacg atgacgataa gatggcccca aagaagaagc ggaaggtcgg tatccacgga
     4501 gtcccagcag ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc
     4561 tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac
     4621 accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa
     4681 acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac
     4741 cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc
     4801 ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc
     4861 atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac
     4921 ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc
     4981 ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac
     5041 aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag
     5101 gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc
     5161 aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg
     5221 ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac
     5281 ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac
     5341 ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc
     5401 gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg
     5461 agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct
     5521 ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac
     5581 ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag
     5641 cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac
     5701 ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga
     5761 gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg
     5821 gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg
     5881 ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac
     5941 ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac
     6001 ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac
     6061 ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc
     6121 gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg
     6181 aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc
     6241 gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg
     6301 ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa
     6361 gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa
     6421 acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc
     6481 ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag
     6541 acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc
     6601 cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc
     6661 gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc
     6721 ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag
     6781 aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc
     6841 cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa
     6901 gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag
     6961 aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat
     7021 gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg
     7081 accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag
     7141 aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc
     7201 gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc
     7261 aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc
     7321 cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc
     7381 ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag
     7441 atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg
     7501 atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac
     7561 gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc
     7621 ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc
     7681 cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc
     7741 cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag
     7801 accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat
     7861 aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc
     7921 accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg
     7981 aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat
     8041 cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag
     8101 ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc
     8161 ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac
     8221 ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg
     8281 tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc
     8341 aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac
     8401 cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat
     8461 ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc
     8521 agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag
     8581 acacggatcg acctgtctca gctgggaggc gacaagcgtc ctgctgctac taagaaagct
     8641 ggtcaagcta agaaaaagaa agctagcgcc acgaacttct ctctgttaaa gcaagcaggc
     8701 gacgtggaag aaaaccccgg tcccgtgagc aagggcgagg aggataacat ggccatcatc
     8761 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
     8821 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
     8881 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
     8941 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
     9001 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     9061 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     9121 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag
     9181 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
     9241 gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag
     9301 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac
     9361 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag
     9421 ctgtacaagt gacgcgttaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     9481 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     9541 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     9601 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     9661 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     9721 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     9781 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     9841 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     9901 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     9961 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
    10021 gggccgcctc cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag
    10081 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
    10141 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
    10201 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
    10261 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
    10321 agtgtggaaa atctctagca gtacgtatag tagttcatgt catcttatta ttcagtattt
    10381 ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa
    10441 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
    10501 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc
    10561 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
    10621 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
    10681 ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata
    10741 gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
    10801 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
    10861 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
    10921 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
    10981 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
    11041 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
    11101 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
    11161 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
    11221 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
    11281 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
    11341 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
    11401 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
    11461 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
    11521 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
    11581 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
    11641 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
    11701 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
    11761 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
    11821 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
    11881 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
    11941 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
    12001 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
    12061 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
    12121 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
    12181 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
    12241 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
    12301 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
    12361 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
    12421 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
    12481 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
    12541 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
    12601 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
    12661 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
    12721 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
    12781 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
    12841 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
    12901 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
    12961 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
    13021 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
    13081 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
    13141 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
    13201 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
    13261 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
    13321 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
    13381 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
    13441 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
    13501 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgc
//
