LOCUS       sequence-457334-       13579 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fusion protein Cas9+GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13579)
  AUTHORS   Pulido-Quetglas C, Aparicio-Prat E, Arnan C, Polidori T, Hermoso T, 
            Palumbo E, Ponomarenko J, Guigo R, Johnson R
  TITLE     Scalable Design of Paired CRISPR Guide RNAs for Genomic Deletion.
  JOURNAL   PLoS Comput Biol. 2017 Mar 2;13(3):e1005341. doi: 
            10.1371/journal.pcbi.1005341. eCollection 2017 Mar.
  PUBMED    28253259
REFERENCE   2  (bases 1 to 13579)
  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: "PLoS Comput
            Biol."; date: "2017-03-2"; volume: "13(3):e1005341. doi"; pages: " 
            10.1371/journal.pcbi.1005341. eCollection 2017 Mar"
COMMENT     Sequence Label: pLentiCas9-GFP
FEATURES             Location/Qualifiers
     source          1..13579
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    52..169
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(467..511)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    696..929
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1422..1547
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             1594..1774
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(1792..1990)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1817..1837)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1992..2371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2543..2562
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2637..2741
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2742..2810,2811..3602)
                     /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(2960..2979)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3773..4361
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4262..4281
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4515..4532
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4649..4670
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4685..4690,4691..4708,4709..4715)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4723..4739
                     /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     4728..4750
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4747..4763
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4747..4763
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4812..4933
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4823..4842)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4877..4896
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5063..5437)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(5456..5503)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     rep_origin      5565..5699
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5619..5638)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     5844..5864
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5873..6301)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5983..6004)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6195..6214
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    6347..6571
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6560..6577
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             6603..6783
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     7288..7304
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7289..7305
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    7308..7896
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             7414..7425
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7823..7843
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7921..8319)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(8320..8376)
                     /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             complement(8386..8403)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(join(8404..9114,9115..9117,9118..9120))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKLSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8446..8467)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     8795..8814
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             complement(9127..9150)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(9151..9198)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(9199..13302)
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(9199..13302)
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     regulatory      13299..13308
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(13327..13538)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
ORIGIN
        1 tagctagcta attaaccaaa ctggatctct gctgtccctg taataaaccc gaaaattttg
       61 aatttttgta atttgttttt gtaattcttt agtttgtatg tctgttgcta ttatgtctac
      121 tattctttcc cctgcactgt accccccaat cccccctttt cttttaaaat tgtggatgaa
      181 tactgccatt tgtctgcaga attggcgcac gcagtgccga tccgttcact aatcgaatgg
      241 atctgtctct gtctctctct ccaccttctt cttctattcc ttcgggcctg tcgggtcccc
      301 tcggggttgg gaggtgggtc tgaaacgata atggtgaata tccctgccta actctattca
      361 ctatagaaag tacagcaaaa actattctta aacctaccaa gcctcctact atcattatga
      421 ataattttat ataccacagc caatttgtta tgttaaacca attccacaaa cttgcccatt
      481 tatctaattc caataattct tgttcattct tttcttgctg gttttgcgat tcttcaatta
      541 aggagtgtat taagcttgtg taattgttaa tttctctgtc ccactccatc caggtcgtgt
      601 gattccaaat ctgttccaga gatttattac tccaactagc attccaaggc acagcagtgg
      661 tgcaaatgag ttttccagag caaccccaaa tccccaggag ctgttgatcc tttaggtatc
      721 tttccacagc caggattctt gcctggagct gcttgatgcc ccagactgtg agttgcaaca
      781 gatgctgttg cgcctcaata gccctcagca aattgttctg ctgctgcact ataccagaca
      841 ataattgtct ggcctgtacc gtcagcgtca ttgacgctgc gcccatagtg cttcctgctg
      901 ctcccaagaa cccaaggaac aaagctccta ttcccactgc tcttttttct ctctgcacca
      961 ctcttctctt tgccttggtg ggtgctactc ctaatggttc aatttttact actttatatt
     1021 tatataattc acttctccaa ttgtccctca tatctcctcc tccaggtctg aagatcagcg
     1081 gccgcttgct gtgcggtggt cttacttttg ttttgctctt cctctatctt gtctaaagct
     1141 tccttggtgt cttttatctc tatcctttga tgcacacaat agagggttgc tactgtatta
     1201 tataatgatc taagttcttc tgatcctgtc tgaagggatg gttgtagctg tcccagtatt
     1261 tgtctacagc cttctgatgt ttctaacagg ccaggattaa ctgcgaatcg ttctagctcc
     1321 ctgcttgccc atactatatg ttttaattta tattttttct ttccccctgg ccttaaccga
     1381 attttttccc atcgcgatct aattctcccc cgcttaatac tgacgctctc gcacccatct
     1441 ctctccttct agcctccgct agtcaaaatt tttggcgtac tcaccagtcg ccgcccctcg
     1501 cctcttgccg tgcgcgcttc agcaagccga gtcctgcgtc gagagagctc ctctggtttc
     1561 cctttcgctt tcaagtccct gttcgggcgc cactgctaga gattttccac actgactaaa
     1621 agggtctgag ggatctctag ttaccagagt cacacaacag acgggcacac actacttgaa
     1681 gcactcaagg caagctttat tgaggcttaa gcagtgggtt ccctagttag ccagagagct
     1741 cccaggctca gatctggtct aaccagagag acccagtaca ggcaaaacgc gctgcttata
     1801 tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga
     1861 cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg
     1921 tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa
     1981 aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag
     2041 tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca
     2101 atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt
     2161 aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac
     2221 gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc
     2281 gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat
     2341 tactattaat aactagtcaa taatcaatgt caacgcgtat atctggcccg tacatcgcga
     2401 agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc aattgtcggt caagccttgc
     2461 cttgttgtag cttaaatttt gctcgcgcac tactcagcga cctccaacac acaagcaggg
     2521 agcagatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatagg
     2581 ggatcgggag atctcccgat ccgtcgacgt caggtggcac ttttcgggga aatgtgcgcg
     2641 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2701 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2761 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2821 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2881 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2941 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     3001 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     3061 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     3121 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     3181 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     3241 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     3301 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     3361 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     3421 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     3481 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3541 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3601 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     3661 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3721 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3781 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3841 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3901 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3961 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4021 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4081 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4141 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4201 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4261 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4321 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4381 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4441 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4501 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     4561 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     4621 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     4681 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     4741 ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag gtcgacggta
     4801 tacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     4861 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     4921 caataaacaa gttggggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca
     4981 tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg
     5041 aatcgaaatc tcgtagcacg tgtcagtcct gctcctcggc cacgaagtgc acgcagttgc
     5101 cggccgggtc gcgcagggcg aactcccgcc cccacggctg ctcgccgatc tcggtcatgg
     5161 ccggcccgga ggcgtcccgg aagttcgtgg acacgacctc cgaccactcg gcgtacagct
     5221 cgtccaggcc gcgcacccac acccaggcca gggtgttgtc cggcaccacc tggtcctgga
     5281 ccgcgctgat gaacagggtc acgtcgtccc ggaccacacc ggcgaagtcg tcctccacga
     5341 agtcccggga gaacccgagc cggtcggtcc agaactcgac cgctccggcg acgtcgcgcg
     5401 cggtgagcac cggaacggca ctggtcaact tggccatggt ttagttcctc accttgtcgt
     5461 attatactat gccgatatac tatgccgatg attaattgtc aacacgtgct gatcagatcc
     5521 gaaaatggat atacaagctc ccgggagctt tttgcaaaag cctaggcctc caaaaaagcc
     5581 tcctcactac ttctggaata gctcagaggc agaggcggcc tcggcctctg cataaataaa
     5641 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggac
     5701 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     5761 ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     5821 tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagaa ttaattcgcg
     5881 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     5941 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     6001 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     6061 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     6121 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     6181 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     6241 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     6301 tggggatacc ccctagagcc ccagctggtt ctttccgcct cagaagccat agagcccacc
     6361 gcatccccag catgcctgct attgtcttcc caatcctccc ccttgctgtc ctgccccacc
     6421 ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc ctcattttat
     6481 taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa
     6541 acaacagatg gctggcaact agaaggcaca gtcgaggctg atcagcgggt ttaaacgggc
     6601 cctgctagag attttccaca ctgactaaaa gggtctgagg gatctctagt taccagagtc
     6661 acacaacaga cgggcacaca ctacttgaag cactcaaggc aagctttatt gaggcttaag
     6721 cagtgggttc cctagttagc cagagagctc ccaggctcag atctggtcta accagagaga
     6781 cccagtacag tccggatgca gctctcgggc catgtgatga aatgctaggc ggctgtcaaa
     6841 cctccactct aatacttctc tctccgggtc atccatccca tgcaggctca cagggtgtaa
     6901 caagcgggtg ttctctcctt cattggcttc ttctaccttc tcttgctcaa ctggtactag
     6961 cttgtagcac catccaaagg tcagtggata tctgatccct ggccctggtg tgtagttctg
     7021 ccaatcaggg aagtagcctt gtgtgtggta gatccacaga tcaaggatat cttgtcttcg
     7081 ttgggagtga attagccctt ccagtccccc cttttctttt aaaaagtggc taagatctac
     7141 agctgccttg taagtcattg gtcttaaagg tacctgaggt gtgactggaa aacccacctc
     7201 ctcctcctct tgtgcttcta gccaggcaca atcagcattg gtagctgctg tattgctact
     7261 tgtgattgct ccatgttttt ctaggtctcg aggtcgacgg tatcgatgcg gggaggcggc
     7321 ccaaagggag atccgactcg tctgagggcg aaggcgaaga cgcggaagag gccgcagagc
     7381 cggcagcagg ccgcgggaag gaaggtccgc tggattgagg gccgaaggga cgtagcagaa
     7441 ggacgtcccg cgcagaatcc aggtggcaac acaggcgagc agccaaggaa aggacgatga
     7501 tttccccgac aacaccacgg aattgtcagt gcccaacagc cgagcccctg tccagcagcg
     7561 ggcaaggcag gcggcgatga gttccgccgt ggcaataggg agggggaaag cgaaagtccc
     7621 ggaaaggagc tgacaggtgg tggcaatgcc ccaaccagtg ggggttgcgt cagcaaacac
     7681 agtgcacacc acgccacgtt gcctgacaac gggccacaac tcctcataaa gagacagcaa
     7741 ccaggattta tacaaggagg agaaaatgaa agccatacgg gaagcaatag catgatacaa
     7801 aggcattaaa gcagcgtatc cacatagcgt aaaaggagca acatagttaa gaataccagt
     7861 caatctttca caaattttgt aatccagagg ttgattaccg ataagcttga tatcgaattc
     7921 ttagccctcc cacacataac cagagggcag caattcacga atcccaactg ccgtcggctg
     7981 tccatcactg tccttcacta tggctttgat cccaggatgc agatcgagaa gcacctgtcg
     8041 gcaccgtccg caggggctca agatgcccct gttctcattt ccgatcgcga cgatacaagt
     8101 caggttgcca gctgccgcag cagcagcagt gcccagcacc acgagttctg cacaaggtcc
     8161 cccagtaaaa tgatatacat tgacaccagt gaagatgcgg ccgtcgctag agagagctgc
     8221 gctggcgacg ctgtagtctt cagagatggg gatgctgttg attgtagccg ttgctctttc
     8281 aatgagggtg gattcttctt gagacaaagg cttggccatc ggtccaggat tctcttcgac
     8341 atctccggct tgtttcagca gagagaagtt tgttgcgccg gatccgtggt gatggtgatg
     8401 atgcttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag
     8461 gaccatgtga tcgcgcttct cgttggggtc tttgctcagg gcggactggg tgctcaggta
     8521 gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc
     8581 ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc ttgaagttca ccttgatgcc
     8641 gttcttctgc ttgtcggcca tgatatagac gttgtggctg ttgtagttgt actccagctt
     8701 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     8761 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
     8821 gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg
     8881 cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg tggtcacgag
     8941 ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc
     9001 gtaggtggca tcgccctcgc cctcgccgga cacgctgagc ttgtggccgt ttacgtcgcc
     9061 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat
     9121 ggatccctta tcgtcatcgt ctttgtaatc tttcttcttc ttagcctgtc cagccttctt
     9181 tgtggcggca ggtcgcttgt cgcctcccag ctgagacagg tcgatccgtg tctcgtacag
     9241 gccggtgatg ctctggtgga tcagggtggc gtccagcacc tctttggtgc tggtgtacct
     9301 cttccggtcg atggtggtgt caaagtactt gaaggcggca ggggctccca gattggtcag
     9361 ggtaaacagg tggatgatat tctcggcctg ctctctgatg ggcttatccc ggtgcttgtt
     9421 gtaggcggac agcactttgt ccagattagc gtcggccagg atcactctct tggagaactc
     9481 gctgatctgc tcgatgatct cgtccaggta gtgcttgtgc tgttccacaa acagctgttt
     9541 ctgctcatta tcctcggggg agcccttcag cttctcatag tggctggcca ggtacaggaa
     9601 gttcacatat ttggagggca gggccagttc gtttcccttc tgcagttcgc cggcagaggc
     9661 cagcattctc ttccggccgt tttccagctc gaacagggag tacttaggca gcttgatgat
     9721 caggtccttt ttcacttctt tgtagccctt ggcttccaga aagtcgatgg gattcttctc
     9781 gaagctgctt ctttccatga tggtgatccc cagtagctct ttcacactct tcagtttctt
     9841 ggacttgccc ttttccactt tggccaccac cagcacagaa taggccacgg tggggctgtc
     9901 gaagccgccg tacttcttag ggtcccagtc cttctttctg gcgatcagct tatcgctgtt
     9961 cctcttgggc aggatagact ctttgctgaa gccgcctgtc tgcacctcgg tctttttcac
    10021 gatattcact tggggcatgc tcagcacttt ccgcacggtg gcaaaatccc ggcccttatc
    10081 ccacacgatc tccccggttt cgccgtttgt ctcgatcaga ggccgcttcc ggatctcgcc
    10141 gttggccagg gtaatctcgg tcttgaaaaa gttcatgatg ttgctgtaga agaagtactt
    10201 ggcggtagcc ttgccgattt cctgctcgct cttggcgatc atcttccgca cgtcgtacac
    10261 cttgtagtcg ccgtacacga actcgctttc cagcttaggg tactttttga tcagggcggt
    10321 tcccacgacg gcgttcaggt aggcgtcgtg ggcgtggtgg tagttgttga tctcgcgcac
    10381 tttgtaaaac tggaaatcct tccggaaatc ggacaccagc ttggacttca gggtgatcac
    10441 tttcacttcc cggatcagct tgtcattctc gtcgtactta gtgttcatcc gggagtccag
    10501 gatctgtgcc acgtgctttg tgatctgccg ggtttccacc agctgtctct tgatgaagcc
    10561 ggccttatcc agttcgctca ggccgcctct ctcggccttg gtcagattgt cgaactttct
    10621 ctgggtaatc agcttggcgt tcagcagctg ccgccagtag ttcttcatct tcttcacgac
    10681 ctcttcggag ggcacgttgt cgctcttgcc ccggttcttg tcgcttctgg tcagcacctt
    10741 gttgtcgatg gagtcgtcct tcagaaagct ctgaggcacg atatggtcca catcgtagtc
    10801 ggacagccgg ttgatgtcca gttcctggtc cacgtacata tcccgcccat tctgcaggta
    10861 gtacaggtac agcttctcgt tctgcagctg ggtgttttcc acggggtgtt ctttcaggat
    10921 ctggctgccc agctctttga tgccctcttc gatccgcttc attctctcgc ggctgttctt
    10981 ctgtcccttc tgggtggtct ggttctctct ggccatttcg atcacgatgt tctcgggctt
    11041 gtgccggccc atcactttca cgagctcgtc caccaccttc actgtctgca ggatgccctt
    11101 cttaatggcg gggctgccgg ccagattggc aatgtgctcg tgcaggctat cgccctggcc
    11161 ggacacctgg gctttctgga tgtcctcttt aaaggtcagg ctgtcgtcgt ggatcagctg
    11221 catgaagttt ctgttggcga agccgtcgga cttcaggaaa tccaggattg tcttgccgga
    11281 ctgcttgtcc cggatgccgt tgatcagctt ccggctcagc ctgccccagc cggtgtatct
    11341 ccgccgcttc agctgcttca tcactttgtc gtcgaacagg tgggcatagg ttttcagccg
    11401 ttcctcgatc atctctctgt cctcaaacag tgtcagggtc agcacgatat cttccagaat
    11461 gtcctcgttt tcctcattgt ccaggaagtc cttgtccttg ataattttca gcagatcgtg
    11521 gtatgtgccc agggaggcgt tgaaccgatc ttccacgccg gagatttcca cggagtcgaa
    11581 gcactcgatt ttcttgaagt agtcctcttt cagctgcttc acggtcactt tccggttggt
    11641 cttgaacagc aggtccacga tggccttttt ctgctcgccg ctcaggaagg cgggctttct
    11701 cattccctcg gtcacgtatt tcactttggt cagctcgtta tacacggtga agtactcgta
    11761 cagcaggctg tgcttgggca gcaccttctc gttgggcagg ttcttatcga agttggtcat
    11821 ccgctcgatg aagctctggg cggaagcgcc cttgtccacc acttcctcga agttccaggg
    11881 ggtgatggtt tcctcgctct ttctggtcat ccaggcgaat ctgctgtttc ccctggccag
    11941 agggcccacg tagtagggga tgcggaaggt caggatcttc tcgatctttt cccggttgtc
    12001 cttcaggaat gggtaaaaat cttcctgccg ccgcagaatg gcgtgcagct ctcccaggtg
    12061 gatctggtgg gggatgctgc cgttgtcgaa ggtccgctgc ttccgcagca ggtcctctct
    12121 gttcagcttc acgagcagtt cctcggtgcc gtccatcttt tccaggatgg gcttgatgaa
    12181 cttgtagaac tcttcctggc tggctccgcc gtcaatgtag ccggcgtagc cgttcttgct
    12241 ctggtcgaag aaaatctctt tgtacttctc aggcagctgc tgccgcacga gagctttcag
    12301 cagggtcagg tcctggtggt gctcgtcgta tctcttgatc atagaggcgc tcaggggggc
    12361 cttggtgatc tcggtgttca ctctcaggat gtcgctcagc aggatggcgt cggacaggtt
    12421 cttggcggcc agaaacaggt cggcgtactg gtcgccgatc tgggccagca ggttgtccag
    12481 gtcgtcgtcg taggtgtcct tgctcagctg cagtttggca tcctcggcca ggtcgaagtt
    12541 gctcttgaag ttgggggtca ggcccaggct cagggcaatc aggtttccga acaggccatt
    12601 cttcttctcg ccgggcagct gggcgatcag attttccagc cgtctgctct tgctcagtct
    12661 ggcagacagg atggccttgg cgtccacgcc gctggcgttg atggggtttt cctcgaacag
    12721 ctggttgtag gtctgcacca gctggatgaa cagcttgtcc acgtcgctgt tgtcggggtt
    12781 caggtcgccc tcgatcagga agtggccccg gaacttgatc atgtgggcca gggccagata
    12841 gatcagccgc aggtcggcct tgtcggtgct gtccaccagt ttctttctca ggtggtagat
    12901 ggtggggtac ttctcgtggt aggccacctc gtccacgatg ttgccgaaga tggggtgccg
    12961 ctcgtgcttc ttatcctctt ccaccaggaa ggactcttcc agtctgtgga agaagctgtc
    13021 gtccaccttg gccatctcgt tgctgaagat ctcttgcaga tagcagatcc ggttcttccg
    13081 tctggtgtat cttcttctgg cggttctctt cagccgggtg gcctcggctg tttcgccgct
    13141 gtcgaacagc agggctccga tcaggttctt cttgatgctg tgccggtcgg tgttgcccag
    13201 caccttgaat ttcttgctgg gcaccttgta ctcgtcggtg atcacggccc agcccacaga
    13261 gttggtgccg atgtccaggc cgatgctgta cttcttgtcc atggtggcag cgctctagaa
    13321 ccggtcctgt gttctggcgg caaacccgtt gcgaaaaaga acgttcacgg cgactactgc
    13381 acttatatac ggttctcccc caccctcggg aaaaaggcgg agccagtaca cgacatcact
    13441 ttcccagttt accccgcgcc accttctcta ggcaccggat caattgccga cccctccccc
    13501 caacttctcg gggactgtgg gcgatgtgcg ctctgcccac tgacgggcac cggagccaat
    13561 tcccactcct ttcaagacc
//
