LOCUS       sequence-230638-       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  JOURNAL   Unpublished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            "Unpublished"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(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        complement(2652..2727)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(2749..2997)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(2998..3014)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        complement(3075..3150)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(3172..3420)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(3421..3437)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3515..3726
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3745..3754
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3751..7854
                     /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             3751..7854
                     /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             7855..7902
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7903..7926
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7936..7992
                     /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             7993..8589
                     /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     8486..8506
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8605..9193
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8658..8678)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9076..9087)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9265..9498
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9524..9541)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9530..9754
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9800..10228
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9887..9906)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10097..10118
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10237..10257)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10242..10571
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10422..10557
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10484..10503
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10619..10666
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10685..11059
                     /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    11189..11310
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11226..11245)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11280..11299
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11359..11375)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11372..11394)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11383..11399
                     /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(11407..11413,11414..11431,11432..11437))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11452..11473
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11590..11607)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11761..12349)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11841..11860)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12520..13311,13312..13380))
                     /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     13143..13162
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13381..13485)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta actcgacaga tctcgtctca catgaggaaa cagctatgac
     2641 catgaaaaaa agcaccgact cggtgccact ttttcaagtt gataacggac tagccttatt
     2701 ttaacttgct atttctagct ctaaaacggg tcccgggatt cttgagccgg tgtttcgtcc
     2761 tttccacaag atatataaag ccaagaaatc gaaatacttt caagttacgg taagcatatg
     2821 atagtccatt ttaaaacata attttaaaac tgcaaactac ccaagaaatt attactttct
     2881 acgtcacgta ttttgtacta atatctttgt gtttacagtc aaattaattc caattatctc
     2941 tctaacagcc ttgtatcgta tatgcaaata tgaaggaatc atgggaaata ggccctcact
     3001 ggccgtcgtt ttacagggtg agacgctcga ggatctcgtc tcacatgagg aaacagctat
     3061 gaccatgaaa aaaagcaccg actcggtgcc actttttcaa gttgataacg gactagcctt
     3121 attttaactt gctatttcta gctctaaaac cgaattatat tctcaacccg cggtgtttcg
     3181 tcctttccac aagatatata aagccaagaa atcgaaatac tttcaagtta cggtaagcat
     3241 atgatagtcc attttaaaac ataattttaa aactgcaaac tacccaagaa attattactt
     3301 tctacgtcac gtattttgta ctaatatctt tgtgtttaca gtcaaattaa ttccaattat
     3361 ctctctaaca gccttgtatc gtatatgcaa atatgaagga atcatgggaa ataggccctc
     3421 actggccgtc gttttacagg gtgagacgct cgaggatccg aattcgctag ctaggtcttg
     3481 aaaggagtgg gaattggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     3541 cccgagaagt tggggggagg ggtcggcaat tgatccggtg cctagagaag gtggcgcggg
     3601 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     3661 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     3721 acacaggacc ggttctagag cgctgccacc atggacaaga agtacagcat cggcctggac
     3781 atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag
     3841 aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc
     3901 ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga
     3961 agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg
     4021 gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat
     4081 aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag
     4141 aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac
     4201 ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc
     4261 gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag
     4321 acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc
     4381 atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc
     4441 ggcgagaaga agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc
     4501 aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc
     4561 tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt
     4621 ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc
     4681 gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag
     4741 gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt
     4801 ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa
     4861 gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc
     4921 gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc
     4981 ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac
     5041 ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac
     5101 tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag
     5161 gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc
     5221 ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag
     5281 cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg
     5341 accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac
     5401 ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag
     5461 aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc
     5521 ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag
     5581 gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag
     5641 atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag
     5701 ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc
     5761 cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac
     5821 agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa
     5881 gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc
     5941 cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg
     6001 atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc
     6061 cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag
     6121 ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag
     6181 ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc
     6241 aaccggctgt ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac
     6301 tccatcgaca acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg
     6361 ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag
     6421 ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa
     6481 ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac
     6541 gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc
     6601 cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc
     6661 cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac
     6721 gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac
     6781 ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc
     6841 aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt
     6901 accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg
     6961 gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc
     7021 caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc
     7081 ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag
     7141 tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa
     7201 aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa
     7261 agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg
     7321 aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg
     7381 aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc
     7441 aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag
     7501 gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc
     7561 gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg
     7621 ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc
     7681 cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc
     7741 atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag
     7801 agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg cgacaagcga
     7861 cctgccgcca caaagaaggc tggacaggct aagaagaaga aagattacaa agacgatgac
     7921 gataagggat ccggcgcaac aaacttctct ctgctgaaac aagccggaga tgtcgaagag
     7981 aatcctggac cgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     8041 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     8101 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     8161 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     8221 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     8281 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     8341 cccaaggagc ccgcgtggtt cctggccacc gtcggagtct cgcccgacca ccagggcaag
     8401 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     8461 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     8521 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     8581 ggtgcctgaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     8641 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     8701 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     8761 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     8821 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     8881 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     8941 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     9001 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     9061 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     9121 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     9181 gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     9241 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     9301 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     9361 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     9421 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     9481 tgtggaaaat ctctagcagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     9541 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     9601 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     9661 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     9721 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     9781 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     9841 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     9901 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     9961 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
    10021 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
    10081 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
    10141 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
    10201 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
    10261 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
    10321 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
    10381 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
    10441 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
    10501 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
    10561 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt
    10621 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
    10681 aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc
    10741 ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc
    10801 cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc
    10861 ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc
    10921 ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga
    10981 gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt
    11041 ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga
    11101 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
    11161 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
    11221 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
    11281 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
    11341 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
    11401 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
    11461 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
    11521 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
    11581 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
    11641 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
    11701 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
    11761 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
    11821 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
    11881 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
    11941 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
    12001 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
    12061 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
    12121 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
    12181 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
    12241 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
    12301 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
    12361 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
    12421 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
    12481 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
    12541 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
    12601 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
    12661 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
    12721 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
    12781 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
    12841 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
    12901 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
    12961 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
    13021 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
    13081 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
    13141 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
    13201 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
    13261 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
    13321 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
    13381 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
    13441 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
    13501 agtgccacct gac
//
