LOCUS       sequence-166526-       14229 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus vector to express guideRNA and dCas9 with puro resistant 
            gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14229)
  AUTHORS   Murakami K, Sasaki H, Nishiyama A, Kurotaki D, Kawase W, Ban T, 
            Nakabayashi J, Kanzaki S, Sekita Y, Nakajima H, Ozato K, Kimura T, 
            Tamura T
  TITLE     A RUNX-CBFbeta-driven enhancer directs the Irf8 dose-dependent 
            lineage choice between DCs and monocytes.
  JOURNAL   Nat Immunol. 2021 Mar;22(3):301-311. doi: 
            10.1038/s41590-021-00871-y. Epub 2021 Feb 18.
  PUBMED    33603226
REFERENCE   2  (bases 1 to 14229)
  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"; doi: 
            "10.1038/s41590-021-00871-y"; journalName: "Nat Immunol"; date: 
            "2021-03"; volume: "22"; issue: "3"; pages: "301-311"
COMMENT     Sequence Label: CSII-U6-gRNA-CBh-3xFLAG-PA-dCas9-P2A-Puro
FEATURES             Location/Qualifiers
     source          1..14229
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        616..814
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             832..1012
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1059..1184
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1681..1914
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2099..2143
                     /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    2417..2534
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2673..2921
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2673..2693
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2844..2863
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2940..3015
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        3112..3397
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        3399..3676
                     /label=chicken beta-actin promoter
     intron          3677..3905
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      3917..3926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3926..3991
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4034..4054
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4079..8179
                     /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
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNMPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             8180..8227
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8237..8293
                     /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             8294..8893
                     /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(8294..8313)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8790..8810
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8923..9511
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8976..8996)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9394..9405)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10030..10210
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10236..10253)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10242..10466
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10512..10940
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10599..10618)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10809..10830
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10949..10969)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10954..11284
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11135..11270
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11197..11216
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11332..11379
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11398..11772
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    11902..12023
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11939..11958)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11993..12012
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12072..12088)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12072..12088)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12085..12107)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12096..12112
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(12120..12126,12127..12144,12145..12150))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12165..12186
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12303..12320)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12474..13062)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12554..12573)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13233..14024,14025..14093))
                     /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     13856..13875
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14094..14198)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc
      841 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      901 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      961 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     1021 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     1081 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     1141 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     1201 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     1261 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     1321 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     1381 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     1441 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     1501 gtaagaccac cgcacagcaa gcggccggcc gctgatcttc agacctggag gaggagatat
     1561 gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg
     1621 agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat
     1681 aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat
     1741 gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt
     1801 gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca
     1861 gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat
     1921 ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag
     1981 taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat
     2041 taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa
     2101 gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat
     2161 aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt
     2221 aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt
     2281 atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga
     2341 agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat ctacaaatgg
     2401 cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa
     2461 gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa
     2521 aaattcaaaa ttttcgggtt tattacaggg acagcagaaa ttcactttga attaattcaa
     2581 gcttcgtgag gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
     2641 aagttggggg gaggggtcgg caattgaacc gggagggcct atttcccatg attccttcat
     2701 atttgcatat acgatacaag gctgttagag agataattgg aattaatttg actgtaaaca
     2761 caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag
     2821 ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg
     2881 atttcttggc tttatatatc ttgtggaaag gacgaaacac cgggtcttcg agaagacctg
     2941 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
     3001 gcaccgagtc ggtgcttttt tgttttagag ctagaaatag caagttaaaa taaggctagt
     3061 ccgtttttag cgcgtgcgcc aattctgcag acaaatggct ctagaggtac ccgttacata
     3121 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     3181 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     3241 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     3301 cggtaaatgg cccgcctggc attgtgccca gtacatgacc ttatgggact ttcctacttg
     3361 gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg
     3421 cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat ttatttttta
     3481 attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgcgccag gcggggcggg
     3541 gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg
     3601 cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg
     3661 aagcgcgcgg cgggcgggag tcgctgcgac gctgccttcg ccccgtgccc cgctccgccg
     3721 ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc
     3781 gggacggccc ttctcctccg ggctgtaatt agctgagcaa gaggtaaggg tttaagggat
     3841 ggttggttgg tggggtatta atgtttaatt acctggagca cctgcctgaa atcacttttt
     3901 ttcaggttgg accggtgcca ccatggacta taaggaccac gacggagact acaaggatca
     3961 tgatattgat tacaaagacg atgacgataa gggcgttgcc atgccaggtg ccgaagatga
     4021 tgtggtgatg gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccga
     4081 caagaagtac agcatcggcc tggccatcgg caccaactct gtgggctggg ccgtgatcac
     4141 cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag
     4201 catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac
     4261 ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct
     4321 gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga
     4381 agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat
     4441 cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact
     4501 ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat
     4561 caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga
     4621 caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa
     4681 cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct
     4741 ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat
     4801 tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc
     4861 caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat
     4921 cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct
     4981 gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat
     5041 caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca
     5101 gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta
     5161 cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa
     5221 gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca
     5281 gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat
     5341 tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa
     5401 gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt
     5461 cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt
     5521 ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct
     5581 gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa
     5641 cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg
     5701 cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa
     5761 gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg
     5821 cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa
     5881 ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac
     5941 cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct
     6001 gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct
     6061 gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt
     6121 cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct
     6181 gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga
     6241 gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa
     6301 ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga
     6361 aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa
     6421 gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga
     6481 aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat
     6541 gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg acgctatcgt
     6601 gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa
     6661 gaaccggggc aagagcgaca acatgccctc cgaagaggtc gtgaagaaga tgaagaacta
     6721 ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa
     6781 ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt
     6841 ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa
     6901 gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct
     6961 ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca
     7021 ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc
     7081 taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat
     7141 cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat
     7201 catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct
     7261 gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac
     7321 cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac
     7381 aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag
     7441 aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc
     7501 tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga
     7561 gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct
     7621 ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc
     7681 cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa
     7741 gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta
     7801 tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca
     7861 caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct
     7921 ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat
     7981 cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc
     8041 cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt
     8101 gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct
     8161 gtctcagctg ggaggcgaca aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa
     8221 gaaaaaggga tccggagcta ctaacttcag cctgctgaag caggctggag acgtggagga
     8281 gaaccctgga cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     8341 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     8401 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     8461 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     8521 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     8581 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     8641 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     8701 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     8761 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     8821 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     8881 gcccggtgcc tgagttaaca tcgagggatc aagcttatcg ataatcaacc tctggattac
     8941 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     9001 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     9061 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     9121 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     9181 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     9241 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     9301 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
     9361 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     9421 tcccgcggcc tgctgccggc tctgcggcct cttccgcgta ttcgccttcg ccctcagacg
     9481 agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgagacct ggaaaaacat
     9541 ggagcaatca caagtagcaa cacagcagct accaatgctg cttgtgcctg gctagaagca
     9601 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
     9661 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     9721 attcactccc aacgatgaca agatatcctt gatctgtgga tctaccacac acaaggctac
     9781 ttccctgatt ggcagaacta cacaccaggg ccagggatca gatatccact gacctttgga
     9841 tggtgctaca agctagtacc agttgagcaa gagaaggtag aagaagccaa tgaaggagag
     9901 aacacccgct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagta
     9961 ttagagtgga ggtttgacag ccgcctagca tttcatcaca tggcccgaga gctgcatccg
    10021 gactgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
    10081 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
    10141 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
    10201 atctctagca gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
    10261 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
    10321 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
    10381 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggatgaca atagcaggca
    10441 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
    10501 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
    10561 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
    10621 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
    10681 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
    10741 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
    10801 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
    10861 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
    10921 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
    10981 ccaggctccc caggcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
    11041 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
    11101 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
    11161 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
    11221 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
    11281 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
    11341 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
    11401 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag
    11461 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg
    11521 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac
    11581 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc
    11641 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg
    11701 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag
    11761 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
    11821 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
    11881 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
    11941 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
    12001 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
    12061 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
    12121 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
    12181 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
    12241 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
    12301 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
    12361 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
    12421 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
    12481 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
    12541 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
    12601 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
    12661 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
    12721 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
    12781 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
    12841 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
    12901 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
    12961 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
    13021 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
    13081 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
    13141 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
    13201 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
    13261 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
    13321 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
    13381 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
    13441 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
    13501 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
    13561 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
    13621 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
    13681 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
    13741 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
    13801 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
    13861 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
    13921 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
    13981 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
    14041 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
    14101 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
    14161 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
    14221 cctgacgtc
//
