LOCUS       sequence-211400-       11203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for expression of Cas9 in mammalian cells (codon 
            optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11203)
  AUTHORS   Zafra MP, Schatoff EM, Katti A, Foronda M, Breinig M, Schweitzer AY,
            Simon A, Han T, Goswami S, Montgomery E, Thibado J, Kastenhuber ER, 
            Sanchez-Rivera FJ, Shi J, Vakoc CR, Lowe SW, Tschaharganeh DF, Dow 
            LE
  TITLE     Optimized base editors enable efficient editing in cells, organoids 
            and mice.
  JOURNAL   Nat Biotechnol. 2018 Jul 3. pii: nbt.4194. doi: 10.1038/nbt.4194.
  PUBMED    29969439
REFERENCE   2  (bases 1 to 11203)
  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: "Nat 
            Biotechnol. 2018 Jul 3. pii: nbt.4194. doi: 10.1038/nbt.4194."
COMMENT     Sequence Label: pLenti-Cas9-P2A-Puro
FEATURES             Location/Qualifiers
     source          1..11203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4..230
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             231..411
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    458..583
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1076..1309
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1494..1538
                     /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    1722..1839
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1908..2119
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2139..2148
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2148..2171
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2178..2198
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2223..6323
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             2223..6323
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6324..6371
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6387..6443
                     /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             6444..7043
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6444..6463)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6940..6960
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7059..7647
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7112..7132)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7530..7541)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7719..7952
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8030..8151
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8067..8086)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8121..8140
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8191..8326
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8346..8365)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8347..8365)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8375..8392)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8375..8391)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8384..8406)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8533..8988
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8620..8639)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8830..8851
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9014..9118
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9119..9187,9188..9979)
                     /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(9337..9356)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10150..10738
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10639..10658
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10892..10909
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11026..11047
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11062..11067,11068..11085,11086..11092)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11100..11116
                     /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     11105..11127
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11124..11140
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11124..11140
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11159..11179
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11161..11179
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 taatgtagtc ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg agttagcaac
       61 atgccttaca aggagagaaa aagcaccgtg catgccgatt ggtggaagta aggtggtacg
      121 atcgtgcctt attaggaagg caacagacgg gtctgacatg gattggacga accactgaat
      181 tgccgcattg cagagatatt gtatttaagt gcctagctcg atacataaac gggtctctct
      241 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
      301 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
      361 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
      421 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     1441 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     1501 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     1561 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     1621 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     1681 ttcagaccca cctcccaacc ccgaggggac ccagaccaca attttaaaag aaaagggggg
     1741 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
     1801 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
     1861 agagatccac tttggcgccg gctcgagtgg ctccggtgcc cgtcagtggg cagagcgcac
     1921 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag
     1981 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     2041 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     2101 gtttgccgcc agaacacagg tgtcgtgacg cgggatccgc caccatggat tacaaagacg
     2161 atgacgataa gatggcccca aagaagaagc ggaaggtcgg tatccacgga gtcccagcag
     2221 ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga
     2281 tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc
     2341 acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg
     2401 ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct
     2461 atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac
     2521 tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca
     2581 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga
     2641 aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca
     2701 tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg
     2761 tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca
     2821 tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac
     2881 ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc
     2941 tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg
     3001 atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc
     3061 agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc
     3121 tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta
     3181 tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc
     3241 agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg
     3301 gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg
     3361 aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga
     3421 agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg
     3481 ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg
     3541 agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca
     3601 gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag
     3661 tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga
     3721 acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt
     3781 ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga
     3841 gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg
     3901 tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct
     3961 ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta
     4021 tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc
     4081 tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc
     4141 acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca
     4201 ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg
     4261 atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca
     4321 gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc
     4381 acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag
     4441 tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga
     4501 tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa
     4561 tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg
     4621 tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg
     4681 atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata
     4741 tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg
     4801 acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga
     4861 actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga
     4921 ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc
     4981 tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca
     5041 ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca
     5101 agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact
     5161 accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt
     5221 accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga
     5281 tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca
     5341 acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc
     5401 ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg
     5461 ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc
     5521 agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg
     5581 ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct
     5641 attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga
     5701 aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact
     5761 ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt
     5821 actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc
     5881 agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc
     5941 actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac
     6001 agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga
     6061 tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc
     6121 ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc
     6181 ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag
     6241 aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg
     6301 acctgtctca gctgggaggc gacaagcgtc ctgctgctac taagaaagct ggtcaagcta
     6361 agaaaaagaa agctagcggc agcggcgcca ccaacttcag cctgctgaag caggccggcg
     6421 acgtggagga gaaccccggc cccatgaccg agtacaagcc cacggtgcgc ctcgccaccc
     6481 gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
     6541 cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct
     6601 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
     6661 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
     6721 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
     6781 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcgga gtctcgcccg
     6841 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     6901 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
     6961 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
     7021 tgacccgcaa gcccggtgcc tgaacgcgtt aagtcgacaa tcaacctctg gattacaaaa
     7081 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     7141 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     7201 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     7261 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     7321 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     7381 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     7441 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     7501 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     7561 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     7621 ggatctccct ttgggccgcc tccccgcgtc gactttaaga ccaatgactt acaaggcagc
     7681 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     7741 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     7801 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     7861 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     7921 acccttttag tcagtgtgga aaatctctag cagtacgtat agtagttcat gtcatcttat
     7981 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat
     8041 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     8101 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     8161 gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     8221 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     8281 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg gacgtaccca
     8341 attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg
     8401 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     8461 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     8521 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     8581 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     8641 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     8701 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     8761 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     8821 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     8881 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     8941 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     9001 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     9061 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     9121 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     9181 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     9241 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     9301 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     9361 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     9421 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     9481 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     9541 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     9601 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     9661 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     9721 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     9781 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     9841 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     9901 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     9961 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
    10021 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
    10081 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
    10141 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
    10201 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
    10261 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
    10321 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
    10381 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
    10441 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
    10501 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
    10561 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
    10621 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
    10681 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
    10741 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
    10801 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
    10861 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
    10921 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
    10981 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
    11041 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
    11101 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc
    11161 aattaaccct cactaaaggg aacaaaagct ggagctgcaa gct
//
