LOCUS       sequence-403311-       11206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9-EDV production plasmid. Expresses the v2 Gag-Cas9 polypeptide 
            (CAG promoter) and sgRNA (human U6 promoter. BsmBI cutsites for 
            spacer insertion).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11206)
  AUTHORS   Hamilton JR, Chen E, Perez BS, Sandoval Espinoza CR, Kang MH, 
            Trinidad M, Ngo W, Doudna JA
  TITLE     In vivo human T cell engineering with enveloped delivery vehicles.
  JOURNAL   Nat Biotechnol. 2024 Jan 11. doi: 10.1038/s41587-023-02085-z.
  PUBMED    38212493
REFERENCE   2  (bases 1 to 11206)
  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. 2024 Jan 11. doi: 10.1038/s41587-023-02085-z."
COMMENT     Sequence Label: pJRH-1179 U6-reci Gag-Cas9 v2
FEATURES             Location/Qualifiers
     source          1..11206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        82..330
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     82..102
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     253..272
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        489..868
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        870..1149
                     /label=chicken beta-actin promoter
     intron          1150..2166
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2090..2112
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2174..2193
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2221..2230
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2227..3726
                     /codon_start=1
                     /gene="gag"
                     /product="gag protein from human immunodeficiency virus 1"
                     /label=HIV-1 gag
                     /translation="MGARASVLSGGELDRWEKIRLRPGGKKKYKLKHIVWASRELERFA
                     VNPGLLETSEGCRQILGQLQPSLQTGSEELRSLYNTVATLYCVHQRIEIKDTKEALDKI
                     EEEQNKSKKKAQQAAADTGHSNQVSQNYPIVQNIQGQMVHQAISPRTLNAWVKVVEEKA
                     FSPEVIPMFSALSEGATPQDLNTMLNTVGGHQAAMQMLKETINEEAAEWDRVHPVHAGP
                     IAPGQMREPRGSDIAGTTSTLQEQIGWMTHNPPIPVGEIYKRWIILGLNKIVRMYSPTS
                     ILDIRQGPKEPFRDYVDRFYKTLRAEQASQEVKNWMTETLLVQNANPDCKTILKALGPG
                     ATLEEMMTACQGVGGPGHKARVLAEAMSQVTNPATIMIQKGNFRNQRKTVKCFNCGKEG
                     HIAKNCRAPRKKGCWKCGKEGHQMKDCTERQANFLGKIWPSHKGRPGNFLQSRPEPTAP
                     PEESFRFGEETTTPSQKQEPIDKELYPLASLRSLFGSDPSSQ"
     CDS             3727..3753
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             3766..3792
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             3805..3831
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             4003..8106
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4003..8106
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8110..8175
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             8194..8214
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8221..8241
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(8254..8273)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    8319..8374
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(8373..8392)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(8735..8751)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8735..8751)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8748..8770)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8759..8775
                     /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(8783..8789,8790..8807,8808..8813))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8828..8849
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8908..9103
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8954..9089
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    9109..9243
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9146..9165)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9200..9219
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9311..9328)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9482..10070)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9562..9581)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10241..11032,11033..11101))
                     /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     10864..10883
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11102..11206)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctggtcg acgggtttat tacagggaca gcagagatcc
       61 agtttggtta attaaggtac cgagggccta tttcccatga ttccttcata tttgcatata
      121 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta
      181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
      241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
      301 ttatatatct tgtggaaagg acgaaacacc ggagacggag gacgacgaac gtctctgttt
      361 aagagctatg ctggaaacag catagcaagt ttaaataagg ctagtccgtt atcaacttga
      421 aaaagtggca ccgagtcggt gctttttttg aattcgctag ctaggtcttg aaaggagtgg
      481 gaattgtcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
      541 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      601 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      661 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      721 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      781 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      841 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
      901 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      961 tgcagcgatg ggggcggggg gggggggggg gggcgcgcgc srggsggggs ggggsggggs
     1021 grggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc
     1081 cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg
     1141 cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg
     1201 cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc
     1261 ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg
     1321 ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg
     1381 ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc
     1441 tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc
     1501 gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg
     1561 cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc
     1621 cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt
     1681 acggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg
     1741 ggcggggcgg ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg
     1801 agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc
     1861 gagagggcgc agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc
     1921 cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg
     1981 ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg
     2041 ggctgtccgc ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc
     2101 tggcgtgtga ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc
     2161 ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc
     2221 gccaccatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg atgggaaaaa
     2281 attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc
     2341 agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga
     2401 caaatactgg gacagctaca accatccctt cagacaggat cagaagaact tagatcatta
     2461 tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag
     2521 gaagctttag acaagataga ggaagagcaa aacaaaagta agaaaaaagc acagcaagca
     2581 gcagctgaca caggacacag caatcaggtc agccaaaatt accctatagt gcagaacatc
     2641 caggggcaaa tggtacatca ggccatatca cctagaactt taaatgcatg ggtaaaagta
     2701 gtagaagaga aggctttcag cccagaagtg atacccatgt tttcagcatt atcagaagga
     2761 gccaccccac aagatttaaa caccatgcta aacacagtgg ggggacatca agcagccatg
     2821 caaatgttaa aagagaccat caatgaggaa gctgcagaat gggatagagt gcatccagtg
     2881 catgcagggc ctattgcacc aggccagatg agagaaccaa ggggaagtga catagcagga
     2941 actactagta cccttcagga acaaatagga tggatgacac ataatccacc tatcccagta
     3001 ggagaaatct ataaaagatg gataatcctg ggattaaata aaatagtaag aatgtatagc
     3061 cctaccagca ttctggacat aagacaagga ccaaaggaac cctttagaga ctatgtagac
     3121 cgattctata aaactctaag agccgagcaa gcttcacaag aggtaaaaaa ttggatgaca
     3181 gaaaccttgt tggtccaaaa tgcgaaccca gattgtaaga ctattttaaa agcattggga
     3241 ccaggagcga cactagaaga aatgatgaca gcatgtcagg gagtgggggg acccggccat
     3301 aaagcaagag ttttggctga agcaatgagc caagtaacaa atccagctac cataatgata
     3361 cagaaaggca attttaggaa ccaaagaaag actgttaagt gtttcaattg tggcaaagaa
     3421 gggcacatag ccaaaaattg cagggcccct aggaaaaagg gctgttggaa atgtggaaag
     3481 gaaggacacc aaatgaaaga ttgtactgag agacaggcta attttttagg gaagatctgg
     3541 ccttcccaca agggaaggcc agggaatttt cttcagagca gaccagagcc aacagcccca
     3601 ccagaagaga gcttcaggtt tggggaagag acaacaactc cctctcagaa gcaggagccg
     3661 atagacaagg aactgtatcc tttagcttcc ctcagatcac tctttggcag cgacccctcg
     3721 tcacaactcc cccctctgga gcgccttact cttggctccc tgcagttgcc cccactggag
     3781 cggctgaccc tggggtcatt gcagttgcct ccactcgaac gactgacgtt gagccagaac
     3841 tatccgattg tgcagaccgg tatgaagcga gcactgccca acaacaccag ctcctctccc
     3901 cagccaaaga agaaaccagg agggtcaggg ggaaagcgag cactgcccaa caacaccagc
     3961 tcctctcccc agccaaagaa gaaaccagga gggtcagggg gaatggacaa gaagtacagc
     4021 atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag
     4081 gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac
     4141 ctgatcggcg ccctgctgtt cgacagcgga gaaacagccg aggccacccg gctgaagaga
     4201 accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc
     4261 agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg
     4321 gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg
     4381 gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc
     4441 gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc
     4501 cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc
     4561 cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg
     4621 gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc
     4681 gcccagctgc ccggcgagaa gaagaatggc ctgttcggca acctgattgc cctgagcctg
     4741 ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg
     4801 agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac
     4861 gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg
     4921 agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac
     4981 gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag
     5041 tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat cgatggcgga
     5101 gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc
     5161 gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac
     5221 aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag
     5281 gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc
     5341 cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc
     5401 agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgcc
     5461 agcgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag
     5521 gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtacaacga gctgaccaaa
     5581 gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaaa
     5641 gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag
     5701 gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg
     5761 ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc
     5821 ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt
     5881 gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa
     5941 gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg
     6001 atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac
     6061 ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag
     6121 gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat
     6181 ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag
     6241 ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag
     6301 aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag
     6361 ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg
     6421 cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag
     6481 gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt
     6541 ctgaaggacg actccatcga taacaaagtg ctgactcgga gcgacaagaa ccggggcaag
     6601 agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg gcgccagctg
     6661 ctgaatgcca agctgattac ccagaggaag ttcgacaatc tgaccaaggc cgagagaggc
     6721 ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag
     6781 atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaac
     6841 gacaaactga tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc
     6901 cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac
     6961 gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc
     7021 gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag
     7081 caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc
     7141 aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac
     7201 ggcgaaacag gcgagatcgt gtgggataag ggccgggact ttgccaccgt gcggaaagtg
     7261 ctgtctatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc
     7321 aaagagtcta tcctgcccaa gaggaacagc gacaagctga tcgccagaaa gaaggactgg
     7381 gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg
     7441 gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc
     7501 accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc
     7561 tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg
     7621 gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg
     7681 gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag
     7741 ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa acactacctg
     7801 gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat
     7861 ctggacaagg tgctgagcgc ctacaacaag cacagagaca agcctatcag agagcaggcc
     7921 gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac
     7981 tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc
     8041 ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga
     8101 ggcgacgccg actataagga ccacgacgga gactacaagg atcatgatat tgattacaaa
     8161 gacgatgacg ataagagcct gggcagcggc tcccccaaga aaaaacgcaa ggtggaagat
     8221 cctaagaaaa agcggaaagt gtaagctagc agatcttttt ccctctgcca aaaattatgg
     8281 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     8341 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     8401 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     8461 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     8521 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     8581 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     8641 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc
     8701 cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     8761 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     8821 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     8881 cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc
     8941 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     9001 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     9061 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt
     9121 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     9181 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     9241 atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     9301 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     9361 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     9421 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     9481 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     9541 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     9601 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     9661 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     9721 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     9781 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     9841 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     9901 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     9961 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
    10021 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
    10081 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
    10141 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
    10201 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
    10261 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
    10321 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
    10381 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
    10441 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
    10501 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
    10561 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
    10621 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
    10681 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
    10741 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
    10801 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
    10861 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
    10921 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
    10981 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
    11041 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
    11101 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
    11161 acatatttga atgtatttag aaaaataaac aaataggggt tccgcg
//
