LOCUS       sequence-237896-       13513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral construct for the expression of Cas9 protein and 
            puromycin resistance from EFS promoter and two guide RNAs targeting 
            for deletion PROX1 start codon sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13513)
  TITLE     Otonkoski plasmids 2018
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 13513)
  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: 
            "Unpublished"
COMMENT     Sequence Label: lentiCRISPRv2_PROX1_1+4
FEATURES             Location/Qualifiers
     source          1..13513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(89..105)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        complement(166..241)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(263..511)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(512..528)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        606..817
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      836..845
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             842..4945
                     /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             842..4945
                     /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             4946..4993
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4994..5017
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5027..5083
                     /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             5084..5680
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     5577..5597
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5696..6284
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5749..5769)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6167..6178)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6356..6589
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6615..6632)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6621..6845
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6891..7319
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6978..6997)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7188..7209
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7328..7348)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7333..7662
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7513..7648
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7575..7594
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7710..7757
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7776..8150
                     /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"
     polyA_signal    8280..8401
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8317..8336)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8371..8390
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8450..8466)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8463..8485)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8474..8490
                     /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)."
     promoter        complement(join(8498..8504,8505..8522,8523..8528))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8543..8564
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8681..8698)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8852..9440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8932..8951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9611..10402,10403..10471))
                     /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     10234..10253
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10472..10576)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10651..10670)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        10842..11221
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11223..11421
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11376..11396
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             11439..11619
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11666..11791
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12284..12517
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12702..12746
                     /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    13044..13161
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        complement(13256..13331)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(join(13353..13513,1..88))
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
ORIGIN
        1 tgtttacagt caaattaatt ccaattatct ctctaacagc cttgtatcgt atatgcaaat
       61 atgaaggaat catgggaaat aggccctcac tggccgtcgt tttacagggt gagacgctcg
      121 aggatctcgt ctcacatgag gaaacagcta tgaccatgaa aaaaagcacc gactcggtgc
      181 cactttttca agttgataac ggactagcct tattttaact tgctatttct agctctaaaa
      241 ccgaattata ttctcaaccc gcggtgtttc gtcctttcca caagatatat aaagccaaga
      301 aatcgaaata ctttcaagtt acggtaagca tatgatagtc cattttaaaa cataatttta
      361 aaactgcaaa ctacccaaga aattattact ttctacgtca cgtattttgt actaatatct
      421 ttgtgtttac agtcaaatta attccaatta tctctctaac agccttgtat cgtatatgca
      481 aatatgaagg aatcatggga aataggccct cactggccgt cgttttacag ggtgagacgc
      541 tcgaggatcc gaattcgcta gctaggtctt gaaaggagtg ggaattggct ccggtgcccg
      601 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
      661 ttgatccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
      721 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
      781 cgttcttttt cgcaacgggt ttgccgccag aacacaggac cggttctaga gcgctgccac
      841 catggacaag aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt
      901 gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg
      961 gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga
     1021 ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg
     1081 ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag
     1141 actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg
     1201 caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa
     1261 gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca
     1321 catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga
     1381 cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc
     1441 catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag
     1501 acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa
     1561 cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga
     1621 ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc
     1681 ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat
     1741 cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc
     1801 tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg
     1861 gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc
     1921 cggctacatt gacggcggag ccagccagga agagttctac aagttcatca agcccatcct
     1981 ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg
     2041 gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca
     2101 cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat
     2161 cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag
     2221 cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga
     2281 agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa
     2341 gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt
     2401 gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct
     2461 gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac
     2521 cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat
     2581 ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat
     2641 tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt
     2701 gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc
     2761 ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg
     2821 caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct
     2881 ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga
     2941 cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct
     3001 gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac
     3061 agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt
     3121 gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag
     3181 aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc
     3241 cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg
     3301 ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca
     3361 tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag
     3421 cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa
     3481 gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct
     3541 gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca
     3601 gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa
     3661 cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc
     3721 caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa
     3781 ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa
     3841 gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa
     3901 gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag
     3961 caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg
     4021 gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt
     4081 tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt
     4141 gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat
     4201 cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc
     4261 ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt
     4321 gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga
     4381 ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa
     4441 gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact
     4501 gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag
     4561 ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga
     4621 acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt
     4681 gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa
     4741 gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc
     4801 ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa
     4861 agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat
     4921 cgacctgtct cagctgggag gcgacaagcg acctgccgcc acaaagaagg ctggacaggc
     4981 taagaagaag aaagattaca aagacgatga cgataaggga tccggcgcaa caaacttctc
     5041 tctgctgaaa caagccggag atgtcgaaga gaatcctgga ccgaccgagt acaagcccac
     5101 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     5161 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     5221 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     5281 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     5341 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     5401 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     5461 cgtcggagtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     5521 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     5581 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     5641 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga acgcgttaag tcgacaatca
     5701 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     5761 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     5821 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     5881 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     5941 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     6001 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     6061 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     6121 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     6181 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     6241 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac tttaagacca
     6301 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     6361 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
     6421 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     6481 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     6541 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag ggcccgttta
     6601 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     6661 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     6721 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     6781 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     6841 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     6901 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     6961 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     7021 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     7081 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     7141 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     7201 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     7261 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     7321 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     7381 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     7441 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     7501 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     7561 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     7621 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     7681 atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg
     7741 gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc
     7801 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt
     7861 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt
     7921 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc
     7981 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg
     8041 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc
     8101 gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac
     8161 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     8221 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     8281 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     8341 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     8401 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     8461 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     8521 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     8581 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     8641 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     8701 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     8761 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     8821 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     8881 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     8941 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     9001 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     9061 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     9121 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     9181 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     9241 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     9301 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     9361 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     9421 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     9481 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     9541 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     9601 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     9661 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     9721 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     9781 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     9841 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     9901 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     9961 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
    10021 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
    10081 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
    10141 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
    10201 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
    10261 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
    10321 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
    10381 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
    10441 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
    10501 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
    10561 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
    10621 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
    10681 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
    10741 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
    10801 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
    10861 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
    10921 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
    10981 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
    11041 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
    11101 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
    11161 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
    11221 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
    11281 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
    11341 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
    11401 gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg ttagaccaga
    11461 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
    11521 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
    11581 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
    11641 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
    11701 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
    11761 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
    11821 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
    11881 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
    11941 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
    12001 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
    12061 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
    12121 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
    12181 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
    12241 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
    12301 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
    12361 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
    12421 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
    12481 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
    12541 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
    12601 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
    12661 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
    12721 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
    12781 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
    12841 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
    12901 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
    12961 acagagacag atccattcga ttagtgaacg gatcggcact gcgtgcgcca attctgcaga
    13021 caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca
    13081 ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa
    13141 attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc agtttggtta
    13201 attaactcga cagatctcgt ctcacatggg gaaacagcta tgaccatgaa aaaaagcacc
    13261 gactcggtgc cactttttca agttgataac ggactagcct tattttaact tgctatttct
    13321 agctctaaaa cgggtcccgg gattcttgag ccggtgtttc gtcctttcca caagatatat
    13381 aaagccaaga aatcgaaata ctttcaagtt acggtaagca tatgatagtc cattttaaaa
    13441 cataatttta aaactgcaaa ctacccaaga aattattact ttctacgtca cgtattttgt
    13501 actaatatct ttg
//
