LOCUS       sequence-235001-       13511 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 13511)
  TITLE     Otonkoski plasmids 2018
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 13511)
  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            "Unpublished"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+3
FEATURES             Location/Qualifiers
     source          1..13511
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        101..174
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     235..251
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        252..500
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        522..597
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    692..809
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(1107..1151)
                     /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    1336..1569
                     /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    2062..2187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             2234..2414
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(2432..2630)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2457..2477)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2632..3011
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3183..3202
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3277..3381
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3382..3450,3451..4242)
                     /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(3600..3619)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4413..5001
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4902..4921
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5155..5172
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5289..5310
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5325..5330,5331..5348,5349..5355)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5363..5379
                     /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     5368..5390
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5387..5403
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    5452..5573
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5463..5482)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5517..5536
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5703..6077)
                     /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(6096..6143)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(6191..6520)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6205..6340
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6259..6278)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6505..6525
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(6534..6962)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6644..6665)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6856..6875
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    7008..7232
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     7221..7238
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             7264..7497
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    7569..8157
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             7675..7686
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     8084..8104
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8173..8769)
                     /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     complement(8256..8276)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             complement(8770..8826)
                     /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(8836..8859)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(8860..8907)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(8908..13011)
                     /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(8908..13011)
                     /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      13008..13017
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(13036..13247)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     primer_bind     13325..13341
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(13342..13511,1..79)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
ORIGIN
        1 ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt
       61 gtggaaagga cgaaacaccg cctgagagca aagcgcgccc gttttagagc tagaaatagc
      121 aagttaaaat aaggctagtc cgttatcctt gaaaaagtgg caccgagtcg gtgctttttt
      181 tcatggtcat agctgtttcc tcatgtgaga cgagatcctc gagcgtctca ccctgtaaaa
      241 cgacggccag tgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
      301 ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata
      361 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
      421 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
      481 tgtggaaagg acgaaacacc ggctcaagaa tcccgggacc cgttttagag ctagaaatag
      541 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
      601 ttttcatggt catagctgtt tcctcatgtg agacgagatc tgtcgagtta attaaccaaa
      661 ctggatctct gctgtccctg taataaaccc gaaaattttg aatttttgta atttgttttt
      721 gtaattcttt agtttgtatg tctgttgcta ttatgtctac tattctttcc cctgcactgt
      781 accccccaat cccccctttt cttttaaaat tgtggatgaa tactgccatt tgtctgcaga
      841 attggcgcac gcagtgccga tccgttcact aatcgaatgg atctgtctct gtctctctct
      901 ccaccttctt cttctattcc ttcgggcctg tcgggtcccc tcggggttgg gaggtgggtc
      961 tgaaacgata atggtgaata tccctgccta actctattca ctatagaaag tacagcaaaa
     1021 actattctta aacctaccaa gcctcctact atcattatga ataattttat ataccacagc
     1081 caatttgtta tgttaaacca attccacaaa cttgcccatt tatctaattc caataattct
     1141 tgttcattct tttcttgctg gttttgcgat tcttcaatta aggagtgtat taagcttgtg
     1201 taattgttaa tttctctgtc ccactccatc caggtcgtgt gattccaaat ctgttccaga
     1261 gatttattac tccaactagc attccaaggc acagcagtgg tgcaaatgag ttttccagag
     1321 caaccccaaa tccccaggag ctgttgatcc tttaggtatc tttccacagc caggattctt
     1381 gcctggagct gcttgatgcc ccagactgtg agttgcaaca gatgctgttg cgcctcaata
     1441 gccctcagca aattgttctg ctgctgcact ataccagaca ataattgtct ggcctgtacc
     1501 gtcagcgtca ttgacgctgc gcccatagtg cttcctgctg ctcccaagaa cccaaggaac
     1561 aaagctccta ttcccactgc tcttttttct ctctgcacca ctcttctctt tgccttggtg
     1621 ggtgctactc ctaatggttc aatttttact actttatatt tatataattc acttctccaa
     1681 ttgtccctca tatctcctcc tccaggtctg aagatcagcg gccgcttgct gtgcggtggt
     1741 cttacttttg ttttgctctt cctctatctt gtctaaagct tccttggtgt cttttatctc
     1801 tatcctttga tgcacacaat agagggttgc tactgtatta tataatgatc taagttcttc
     1861 tgatcctgtc tgaagggatg gttgtagctg tcccagtatt tgtctacagc cttctgatgt
     1921 ttctaacagg ccaggattaa ctgcgaatcg ttctagctcc ctgcttgccc atactatatg
     1981 ttttaattta tattttttct ttccccctgg ccttaaccga attttttccc atcgcgatct
     2041 aattctcccc cgcttaatac tgacgctctc gcacccatct ctctccttct agcctccgct
     2101 agtcaaaatt tttggcgtac tcaccagtcg ccgcccctcg cctcttgccg tgcgcgcttc
     2161 agcaagccga gtcctgcgtc gagagagctc ctctggtttc cctttcgctt tcaagtccct
     2221 gttcgggcgc cactgctaga gattttccac actgactaaa agggtctgag ggatctctag
     2281 ttaccagagt cacacaacag acgggcacac actacttgaa gcactcaagg caagctttat
     2341 tgaggcttaa gcagtgggtt ccctagttag ccagagagct cccaggctca gatctggtct
     2401 aaccagagag acccagtaca ggcaaaacgc gctgcttata tagacctccc accgtacacg
     2461 cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg
     2521 attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg
     2581 tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat
     2641 agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg
     2701 ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat
     2761 atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg
     2821 tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg
     2881 gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa
     2941 ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagtcaa
     3001 taatcaatgt caacgcgtat atctggcccg tacatcgcga agcagcgcaa aacgcctaac
     3061 cctaagcaga ttcttcatgc aattgtcggt caagccttgc cttgttgtag cttaaatttt
     3121 gctcgcgcac tactcagcga cctccaacac acaagcaggg agcagatact ggcttaacta
     3181 tgcggcatca gagcagattg tactgagagt gcaccatagg ggatcgggag atctcccgat
     3241 ccgtcgacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3301 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3361 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3421 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3481 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3541 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3601 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3661 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3721 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3781 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3841 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3901 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     3961 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4021 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4081 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4141 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4201 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4261 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4321 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4381 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4441 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4501 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4561 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4621 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4681 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4741 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4801 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4861 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4921 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4981 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5041 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5101 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     5161 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     5221 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     5281 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     5341 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     5401 gaccatgatt acgccaagct ctagctagag gtcgacggta tacagacatg ataagataca
     5461 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     5521 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttggggtgg
     5581 gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa
     5641 acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtagcacg
     5701 tgtcagtcct gctcctcggc cacgaagtgc acgcagttgc cggccgggtc gcgcagggcg
     5761 aactcccgcc cccacggctg ctcgccgatc tcggtcatgg ccggcccgga ggcgtcccgg
     5821 aagttcgtgg acacgacctc cgaccactcg gcgtacagct cgtccaggcc gcgcacccac
     5881 acccaggcca gggtgttgtc cggcaccacc tggtcctgga ccgcgctgat gaacagggtc
     5941 acgtcgtccc ggaccacacc ggcgaagtcg tcctccacga agtcccggga gaacccgagc
     6001 cggtcggtcc agaactcgac cgctccggcg acgtcgcgcg cggtgagcac cggaacggca
     6061 ctggtcaact tggccatggt ttagttcctc accttgtcgt attatactat gccgatatac
     6121 tatgccgatg attaattgtc aacacgtgct gatcagatcc gaaaatggat atacaagctc
     6181 ccgggagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata
     6241 gctcagaggc agaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg
     6301 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
     6361 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     6421 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     6481 ctggggagcc tggggacttt ccacacccta actgacacac attccacaga attaattcgc
     6541 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     6601 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     6661 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     6721 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     6781 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     6841 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     6901 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     6961 gtggggatac cccctagagc cccagctggt tctttccgcc tcagaagcca tagagcccac
     7021 cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac
     7081 cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta
     7141 ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca
     7201 aacaacagat ggctggcaac tagaaggcac agtcgaggct gatcagcggg tttaaacggg
     7261 ccctgctaga gattttccac actgactaaa agggtctgag ggatctctag ttaccagagt
     7321 cacacaacag acgggcacac actacttgaa gcactcaagg caagctttat tgaggcttaa
     7381 gcagtgggtt ccctagttag ccagagagct cccaggctca gatctggtct aaccagagag
     7441 acccagtaca agcaaaaagc agatcttgtc ttcgttggga gtgaattagc ccttccagtc
     7501 cccccttttc ttttaaaaag tggctaagat ctacagctgc cttgtaagtc attggtctta
     7561 aagtcgacgc ggggaggcgg cccaaaggga gatccgactc gtctgagggc gaaggcgaag
     7621 acgcggaaga ggccgcagag ccggcagcag gccgcgggaa ggaaggtccg ctggattgag
     7681 ggccgaaggg acgtagcaga aggacgtccc gcgcagaatc caggtggcaa cacaggcgag
     7741 cagccaagga aaggacgatg atttccccga caacaccacg gaattgtcag tgcccaacag
     7801 ccgagcccct gtccagcagc gggcaaggca ggcggcgatg agttccgccg tggcaatagg
     7861 gagggggaaa gcgaaagtcc cggaaaggag ctgacaggtg gtggcaatgc cccaaccagt
     7921 gggggttgcg tcagcaaaca cagtgcacac cacgccacgt tgcctgacaa cgggccacaa
     7981 ctcctcataa agagacagca accaggattt atacaaggag gagaaaatga aagccatacg
     8041 ggaagcaata gcatgataca aaggcattaa agcagcgtat ccacatagcg taaaaggagc
     8101 aacatagtta agaataccag tcaatctttc acaaattttg taatccagag gttgattgtc
     8161 gacttaacgc gttcaggcac cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg
     8221 cacctcgacg tcggcggtga cggtgaagcc gagccgctcg tagaagggga ggttgcgggg
     8281 cgcggaggtc tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca ctccggggag
     8341 cacgacggcg ctgcccagac ccttgccctg gtggtcgggc gagactccga cggtggccag
     8401 gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg ccttccatct gttgctgcgc
     8461 ggccagccgg gaaccgctca actcggccat gcgcgggccg atctcggcga acaccgcccc
     8521 cgcttcgacg ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca
     8581 caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg
     8641 ctcgatgtgg cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc
     8701 ggcgagggtg cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt
     8761 gtactcggtc ggtccaggat tctcttcgac atctccggct tgtttcagca gagagaagtt
     8821 tgttgcgccg gatcccttat cgtcatcgtc tttgtaatct ttcttcttct tagcctgtcc
     8881 agccttcttt gtggcggcag gtcgcttgtc gcctcccagc tgagacaggt cgatccgtgt
     8941 ctcgtacagg ccggtgatgc tctggtggat cagggtggcg tccagcacct ctttggtgct
     9001 ggtgtacctc ttccggtcga tggtggtgtc aaagtacttg aaggcggcag gggctcccag
     9061 attggtcagg gtaaacaggt ggatgatatt ctcggcctgc tctctgatgg gcttatcccg
     9121 gtgcttgttg taggcggaca gcactttgtc cagattagcg tcggccagga tcactctctt
     9181 ggagaactcg ctgatctgct cgatgatctc gtccaggtag tgcttgtgct gttccacaaa
     9241 cagctgtttc tgctcattat cctcggggga gcccttcagc ttctcatagt ggctggccag
     9301 gtacaggaag ttcacatatt tggagggcag ggccagttcg tttcccttct gcagttcgcc
     9361 ggcagaggcc agcattctct tccggccgtt ttccagctcg aacagggagt acttaggcag
     9421 cttgatgatc aggtcctttt tcacttcttt gtagcccttg gcttccagaa agtcgatggg
     9481 attcttctcg aagctgcttc tttccatgat ggtgatcccc agcagctctt tcacactctt
     9541 cagtttcttg gacttgccct tttccacttt ggccaccacc agcacagaat aggccacggt
     9601 ggggctgtcg aagccgccgt acttcttagg gtcccagtcc ttctttctgg cgatcagctt
     9661 atcgctgttc ctcttgggca ggatagactc tttgctgaag ccgcctgtct gcacctcggt
     9721 ctttttcacg atattcactt ggggcatgct cagcactttc cgcacggtgg caaaatcccg
     9781 gcccttatcc cacacgatct ccccggtttc gccgtttgtc tcgatcagag gccgcttccg
     9841 gatctcgccg ttggccaggg taatctcggt cttgaaaaag ttcatgatgt tgctgtagaa
     9901 gaagtacttg gcggtagcct tgccgatttc ctgctcgctc ttggcgatca tcttccgcac
     9961 gtcgtacacc ttgtagtcgc cgtacacgaa ctcgctttcc agcttagggt actttttgat
    10021 cagggcggtt cccacgacgg cgttcaggta ggcgtcgtgg gcgtggtggt agttgttgat
    10081 ctcgcgcact ttgtaaaact ggaaatcctt ccggaaatcg gacaccagct tggacttcag
    10141 ggtgatcact ttcacttccc ggatcagctt gtcattctcg tcgtacttag tgttcatccg
    10201 ggagtccagg atctgtgcca cgtgctttgt gatctgccgg gtttccacca gctgtctctt
    10261 gatgaagccg gccttatcca gttcgctcag gccgcctctc tcggccttgg tcagattgtc
    10321 gaactttctc tgggtaatca gcttggcgtt cagcagctgc cgccagtagt tcttcatctt
    10381 cttcacgacc tcttcggagg gcacgttgtc gctcttgccc cggttcttgt cgcttctggt
    10441 cagcaccttg ttgtcgatgg agtcgtcctt cagaaagctc tgaggcacga tatggtccac
    10501 atcgtagtcg gacagccggt tgatgtccag ttcctggtcc acgtacatat cccgcccatt
    10561 ctgcaggtag tacaggtaca gcttctcgtt ctgcagctgg gtgttttcca cggggtgttc
    10621 tttcaggatc tggctgccca gctctttgat gccctcttcg atccgcttca ttctctcgcg
    10681 gctgttcttc tgtcccttct gggtggtctg gttctctctg gccatttcga tcacgatgtt
    10741 ctcgggcttg tgccggccca tcactttcac gagctcgtcc accaccttca ctgtctgcag
    10801 gatgcccttc ttaatggcgg ggctgccggc cagattggca atgtgctcgt gcaggctatc
    10861 gccctggccg gacacctggg ctttctggat gtcctcttta aaggtcaggc tgtcgtcgtg
    10921 gatcagctgc atgaagtttc tgttggcgaa gccgtcggac ttcaggaaat ccaggattgt
    10981 cttgccggac tgcttgtccc ggatgccgtt gatcagcttc cggctcagcc tgccccagcc
    11041 ggtgtatctc cgccgcttca gctgcttcat cactttgtcg tcgaacaggt gggcataggt
    11101 tttcagccgt tcctcgatca tctctctgtc ctcaaacagt gtcagggtca gcacgatatc
    11161 ttccagaatg tcctcgtttt cctcattgtc caggaagtcc ttgtccttga taattttcag
    11221 cagatcgtgg tatgtgccca gggaggcgtt gaaccgatct tccacgccgg agatttccac
    11281 ggagtcgaag cactcgattt tcttgaagta gtcctctttc agctgcttca cggtcacttt
    11341 ccggttggtc ttgaacagca ggtccacgat ggcctttttc tgctcgccgc tcaggaaggc
    11401 gggctttctc attccctcgg tcacgtattt cactttggtc agctcgttat acacggtgaa
    11461 gtactcgtac agcaggctgt gcttgggcag caccttctcg ttgggcaggt tcttatcgaa
    11521 gttggtcatc cgctcgatga agctctgggc ggaagcgccc ttgtccacca cttcctcgaa
    11581 gttccagggg gtgatggttt cctcgctctt tctggtcatc caggcgaatc tgctgtttcc
    11641 cctggccaga gggcccacgt agtaggggat gcggaaggtc aggatcttct cgatcttttc
    11701 ccggttgtcc ttcaggaatg ggtaaaaatc ttcctgccgc cgcagaatgg cgtgcagctc
    11761 tcccaggtgg atctggtggg ggatgctgcc gttgtcgaag gtccgctgct tccgcagcag
    11821 gtcctctctg ttcagcttca cgagcagttc ctcggtgccg tccatctttt ccaggatggg
    11881 cttgatgaac ttgtagaact cttcctggct ggctccgccg tcaatgtagc cggcgtagcc
    11941 gttcttgctc tggtcgaaga aaatctcttt gtacttctca ggcagctgct gccgcacgag
    12001 agctttcagc agggtcaggt cctggtggtg ctcgtcgtat ctcttgatca tagaggcgct
    12061 caggggggcc ttggtgatct cggtgttcac tctcaggatg tcgctcagca ggatggcgtc
    12121 ggacaggttc ttggcggcca gaaacaggtc ggcgtactgg tcgccgatct gggccagcag
    12181 gttgtccagg tcgtcgtcgt aggtgtcctt gctcagctgc agtttggcat cctcggccag
    12241 gtcgaagttg ctcttgaagt tgggggtcag gcccaggctc agggcaatca ggtttccgaa
    12301 caggccattc ttcttctcgc cgggcagctg ggcgatcaga ttttccagcc gtctgctctt
    12361 gctcagtctg gcagacagga tggccttggc gtccacgccg ctggcgttga tggggttttc
    12421 ctcgaacagc tggttgtagg tctgcaccag ctggatgaac agcttgtcca cgtcgctgtt
    12481 gtcggggttc aggtcgccct cgatcaggaa gtggccccgg aacttgatca tgtgggccag
    12541 ggccagatag atcagccgca ggtcggcctt gtcggtgctg tccaccagtt tctttctcag
    12601 gtggtagatg gtggggtact tctcgtggta ggccacctcg tccacgatgt tgccgaagat
    12661 ggggtgccgc tcgtgcttct tatcctcttc caccaggaag gactcttcca gtctgtggaa
    12721 gaagctgtcg tccaccttgg ccatctcgtt gctgaagatc tcttgcagat agcagatccg
    12781 gttcttccgt ctggtgtatc ttcttctggc ggttctcttc agccgggtgg cctcggctgt
    12841 ttcgccgctg tcgaacagca gggctccgat caggttcttc ttgatgctgt gccggtcggt
    12901 gttgcccagc accttgaatt tcttgctggg caccttgtac tcgtcggtga tcacggccca
    12961 gcccacagag ttggtgccga tgtccaggcc gatgctgtac ttcttgtcca tggtggcagc
    13021 gctctagaac cggtcctgtg ttctggcggc aaacccgttg cgaaaaagaa cgttcacggc
    13081 gactactgca cttatatacg gttctccccc accctcggga aaaaggcgga gccagtacac
    13141 gacatcactt tcccagttta ccccgcgcca ccttctctag gcaccggatc aattgccgac
    13201 ccctcccccc aacttctcgg ggactgtggg cgatgtgcgc tctgcccact gacgggcacc
    13261 ggagccaatt cccactcctt tcaagaccta gctagcgaat tcggatcctc gagcgtctca
    13321 ccctgtaaaa cgacggccag tgagggccta tttcccatga ttccttcata tttgcatata
    13381 cgatacaagg ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta
    13441 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
    13501 tgttttaaaa t
//
