LOCUS       sequence-211409-       12491 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lentiviral vector for constitutive expression of FNLS in mammalian 
            cells (codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12491)
  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 12491)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-FNLS-PGK-Puro
FEATURES             Location/Qualifiers
     source          1..12491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(55..74)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        246..625
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        627..825
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     780..800
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             843..1023
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1070..1195
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1692..1925
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2110..2154
                     /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    2452..2569
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2687..2898
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2934..2943
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2943..3008
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3015..3035
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3060..3746
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             join(3795..3818,3819..3821,3822..7895)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             3795..7895
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7908..8156
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             8169..8189
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        8216..8715
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8251..8274)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     8648..8667
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             8736..9335
                     /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(8736..8755)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9232..9252
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9388..9976
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9441..9461)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9859..9870)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9979..9995)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9980..9996)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10505..10685
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(10747..11335)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10827..10846)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11506..12297,12298..12366))
                     /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     12129..12148
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12367..12471)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
       61 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
      121 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
      181 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
      241 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      301 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      361 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      421 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      481 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      541 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      601 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      661 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      721 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      781 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagcgcgt tttgcctgta
      841 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
      901 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
      961 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     1021 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg
     1081 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
     1141 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
     1201 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
     1261 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
     1321 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
     1381 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
     1441 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga
     1501 gcaaaacaaa agtaagacca ccgcacagca agcggccggc cgctgatctt cagacctgga
     1561 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     1621 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     1681 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     1741 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     1801 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     1861 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     1921 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     1981 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
     2041 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
     2101 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
     2161 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
     2221 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
     2281 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa
     2341 ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga
     2401 tcggcactgc gtgcgccaat tctgcagaca aatggcagta ttcatccaca attttaaaag
     2461 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     2521 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
     2581 cagggacagc agagatccag tttggttagt accgggcccg ggtcgactag tgatcagtgt
     2641 gagggagtgt aaagctggtt ttcgagtggc tccagtgccc gtcagtgggc agagcgcaca
     2701 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga
     2761 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     2821 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     2881 tttgccgcca gaacacaggt gtcgtgacgc ggctaactcg aacgctagga tccgccacca
     2941 tggactataa ggaccacgac ggagactaca aggatcatga tattgattac aaagacgatg
     3001 acgataagat ggccccaaag aagaagcgga aggtcggtat ccacggagtc ccagcagcca
     3061 tgagctcaga gactggccca gtggctgtgg accccacatt gagacggcgg atcgagcccc
     3121 atgagtttga ggtattcttc gatccgagag agctccgcaa ggagacctgc ctgctttacg
     3181 aaattaattg ggggggccgg cactccattt ggcgacatac atcacagaac actaacaagc
     3241 acgtcgaagt caacttcatc gagaagttca cgacagaaag atatttctgt ccgaacacaa
     3301 ggtgcagcat tacctggttt ctcagctgga gcccatgcgg cgaatgtagt agggccatca
     3361 ctgaattcct gtcaaggtat ccccacgtca ctctgtttat ttacatcgca aggctgtacc
     3421 accacgctga cccccgcaat cgacaaggcc tgcgggattt gatctcttca ggtgtgacta
     3481 tccaaattat gactgagcag gagtcaggat actgctggag aaactttgtg aattatagcc
     3541 cgagtaatga agcccactgg cctaggtatc cccatctgtg ggtacgactg tacgttcttg
     3601 aactgtactg catcatactg ggcctgcctc cttgtctcaa cattctgaga aggaagcagc
     3661 cacagctgac attctttacc atcgctcttc agtcttgtca ttaccagcga ctgcccccac
     3721 acattctctg ggccaccggg ttgaaaagcg gcagcgagac tcccgggacc tcagagtccg
     3781 ccacacccga aagtgacaag aagtacagca tcggcctggc catcggcacc aactctgtgg
     3841 gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca
     3901 acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg
     3961 aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga
     4021 accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct
     4081 tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc
     4141 ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc
     4201 acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg
     4261 ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg
     4321 acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg
     4381 aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga
     4441 gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc
     4501 tgttcggcaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg
     4561 acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca
     4621 acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt
     4681 ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc
     4741 tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag
     4801 ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga
     4861 acggctacgc cggctacatt gacggcggag ccagccagga agagttctac aagttcatca
     4921 agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg
     4981 acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg
     5041 gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc
     5101 gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca
     5161 ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga
     5221 acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca
     5281 acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt
     5341 acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc
     5401 ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc
     5461 ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact
     5521 ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc
     5581 tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg
     5641 aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga
     5701 aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca
     5761 ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca
     5821 agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga
     5881 tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg
     5941 gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca
     6001 tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg
     6061 agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca
     6121 gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga
     6181 aagaacaccc agtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc
     6241 agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg
     6301 atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc
     6361 tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga
     6421 agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt
     6481 tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca
     6541 tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact
     6601 cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca
     6661 ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg
     6721 agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc
     6781 tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg
     6841 acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact
     6901 tcttctacag caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga
     6961 tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg
     7021 gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa
     7081 agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg
     7141 ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc
     7201 ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac
     7261 tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga
     7321 atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca
     7381 agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg
     7441 ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt
     7501 acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc
     7561 tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct
     7621 ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc
     7681 accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca
     7741 atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca
     7801 ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg
     7861 agacacggat cgacctgtct cagctgggag gcgattcagg cggatctact aatctgtcag
     7921 atattattga aaaggagacc ggtaagcaac tggttatcca ggaatccatc ctcatgctcc
     7981 cagaggaggt ggaagaagtc attgggaaca agccggaaag cgatatactc gtgcacaccg
     8041 cctacgacga gagcaccgac gagaatgtca tgcttctgac tagcgacgcc cctgaataca
     8101 agccttgggc tctggtcata caggatagca acggtgagaa caagattaag atgctctctg
     8161 gtggttctcc caagaagaag aggaaagtct aactaggaat cccgaggcct ctaccgggta
     8221 ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact
     8281 tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa
     8341 ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag
     8401 ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc
     8461 actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc
     8521 agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg
     8581 tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga
     8641 ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca
     8701 tctccgggcc tttcgacctg cagcccaagc ttaccatgac cgagtacaag cccacggtgc
     8761 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     8821 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     8881 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     8941 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     9001 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     9061 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     9121 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     9181 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     9241 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     9301 gcacctggtg catgacccgc aagcccggtg cctgaaaacg ttgttgtttg gggttgaatt
     9361 actcttctag accgcggcgc gcccgataat caacctctgg attacaaaat ttgtgaaaga
     9421 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     9481 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     9541 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     9601 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     9661 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     9721 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     9781 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     9841 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     9901 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     9961 tgggccgcct ccccgcatcg ataccgtcga cctcgatcga gacctagaaa aacatggagc
    10021 aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga
    10081 ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa
    10141 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
    10201 ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
    10261 tgattggcag aactacacac cagggccagg gatcagatat ccactgacct ttggatggtg
    10321 ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag gagagaacac
    10381 ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtattaga
    10441 gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc atccggactg
    10501 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
    10561 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
    10621 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
    10681 tagcagcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
    10741 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
    10801 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
    10861 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
    10921 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
    10981 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
    11041 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
    11101 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
    11161 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
    11221 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
    11281 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
    11341 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
    11401 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
    11461 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
    11521 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
    11581 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
    11641 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
    11701 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
    11761 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
    11821 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
    11881 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
    11941 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
    12001 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
    12061 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
    12121 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
    12181 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
    12241 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
    12301 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
    12361 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
    12421 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
    12481 ccgaaaagtg c
//
