LOCUS       sequence-403306-       11200 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9-EDV production plasmid. Expresses the v2 Gag-Cas9 polypeptide 
            (CAG promoter) and B2M sgRNA (human U6 promoter, 
            5'-GAGTAGCGCGAGCACAGCTA).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11200)
  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 11200)
  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-1345 U6-B2M sgRNA Gag-Cas9 v2
FEATURES             Location/Qualifiers
     source          1..11200
                     /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        484..863
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        865..1143
                     /label=chicken beta-actin promoter
     intron          1144..2160
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2084..2106
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2168..2187
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      2215..2224
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2221..3720
                     /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             3721..3747
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             3760..3786
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             3799..3825
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             3997..8100
                     /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             3997..8100
                     /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             8104..8169
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             8188..8208
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8215..8235
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(8248..8267)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    8313..8368
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(8367..8386)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(8729..8745)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8729..8745)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8742..8764)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8753..8769
                     /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(8777..8783,8784..8801,8802..8807))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8822..8843
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8902..9097
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8948..9083
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    9103..9237
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9140..9159)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9194..9213
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9305..9322)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9476..10064)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9556..9575)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10235..11026,11027..11095))
                     /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     10858..10877
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11096..11200)
                     /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 ggagtagcgc gagcacagct agtttaagag
      361 ctatgctgga aacagcatag caagtttaaa taaggctagt ccgttatcaa cttgaaaaag
      421 tggcaccgag tcggtgcttt ttttgaattc gctagctagg tcttgaaagg agtgggaatt
      481 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      541 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      601 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      661 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      721 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      781 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      841 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      901 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      961 cgatgggggc gggggggggg gggggggcgc gcgccaggcg gggcggggcg gggcgagggg
     1021 cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
     1081 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
     1141 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
     1201 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
     1261 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
     1321 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg
     1381 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag
     1441 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc
     1501 gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt
     1561 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg
     1621 cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg
     1681 cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg
     1741 gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc
     1801 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     1861 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
     1921 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     1981 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
     2041 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     2101 gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag
     2161 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcgccacc
     2221 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgatggga aaaaattcgg
     2281 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
     2341 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
     2401 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     2461 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     2521 ttagacaaga tagaggaaga gcaaaacaaa agtaagaaaa aagcacagca agcagcagct
     2581 gacacaggac acagcaatca ggtcagccaa aattacccta tagtgcagaa catccagggg
     2641 caaatggtac atcaggccat atcacctaga actttaaatg catgggtaaa agtagtagaa
     2701 gagaaggctt tcagcccaga agtgataccc atgttttcag cattatcaga aggagccacc
     2761 ccacaagatt taaacaccat gctaaacaca gtggggggac atcaagcagc catgcaaatg
     2821 ttaaaagaga ccatcaatga ggaagctgca gaatgggata gagtgcatcc agtgcatgca
     2881 gggcctattg caccaggcca gatgagagaa ccaaggggaa gtgacatagc aggaactact
     2941 agtacccttc aggaacaaat aggatggatg acacataatc cacctatccc agtaggagaa
     3001 atctataaaa gatggataat cctgggatta aataaaatag taagaatgta tagccctacc
     3061 agcattctgg acataagaca aggaccaaag gaacccttta gagactatgt agaccgattc
     3121 tataaaactc taagagccga gcaagcttca caagaggtaa aaaattggat gacagaaacc
     3181 ttgttggtcc aaaatgcgaa cccagattgt aagactattt taaaagcatt gggaccagga
     3241 gcgacactag aagaaatgat gacagcatgt cagggagtgg ggggacccgg ccataaagca
     3301 agagttttgg ctgaagcaat gagccaagta acaaatccag ctaccataat gatacagaaa
     3361 ggcaatttta ggaaccaaag aaagactgtt aagtgtttca attgtggcaa agaagggcac
     3421 atagccaaaa attgcagggc ccctaggaaa aagggctgtt ggaaatgtgg aaaggaagga
     3481 caccaaatga aagattgtac tgagagacag gctaattttt tagggaagat ctggccttcc
     3541 cacaagggaa ggccagggaa ttttcttcag agcagaccag agccaacagc cccaccagaa
     3601 gagagcttca ggtttgggga agagacaaca actccctctc agaagcagga gccgatagac
     3661 aaggaactgt atcctttagc ttccctcaga tcactctttg gcagcgaccc ctcgtcacaa
     3721 ctcccccctc tggagcgcct tactcttggc tccctgcagt tgcccccact ggagcggctg
     3781 accctggggt cattgcagtt gcctccactc gaacgactga cgttgagcca gaactatccg
     3841 attgtgcaga ccggtatgaa gcgagcactg cccaacaaca ccagctcctc tccccagcca
     3901 aagaagaaac caggagggtc agggggaaag cgagcactgc ccaacaacac cagctcctct
     3961 ccccagccaa agaagaaacc aggagggtca gggggaatgg acaagaagta cagcatcggc
     4021 ctggacatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc
     4081 agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc
     4141 ggcgccctgc tgttcgacag cggagaaaca gccgaggcca cccggctgaa gagaaccgcc
     4201 agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac
     4261 gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa
     4321 gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac
     4381 cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag
     4441 gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc
     4501 ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg
     4561 gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc
     4621 aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag
     4681 ctgcccggcg agaagaagaa tggcctgttc ggcaacctga ttgccctgag cctgggcctg
     4741 acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag
     4801 gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac
     4861 ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg
     4921 aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac
     4981 caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa
     5041 gagattttct tcgaccagag caagaacggc tacgccggct acatcgatgg cggagccagc
     5101 caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa
     5161 ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc
     5221 agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat
     5281 ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc
     5341 ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag
     5401 agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgccagcgcc
     5461 cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg
     5521 cccaagcaca gcctgctgta cgagtacttc accgtgtaca acgagctgac caaagtgaaa
     5581 tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaagccatc
     5641 gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac
     5701 ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac
     5761 gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac
     5821 aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac
     5881 agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg
     5941 aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac
     6001 ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc
     6061 gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc
     6121 cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc
     6181 ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg
     6241 aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag
     6301 accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc
     6361 aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac
     6421 gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg
     6481 gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag
     6541 gacgactcca tcgataacaa agtgctgact cggagcgaca agaaccgggg caagagcgac
     6601 aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcgcca gctgctgaat
     6661 gccaagctga ttacccagag gaagttcgac aatctgacca aggccgagag aggcggcctg
     6721 agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca
     6781 aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga gaacgacaaa
     6841 ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag
     6901 gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac
     6961 ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc
     7021 gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa
     7081 atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc
     7141 gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa
     7201 acaggcgaga tcgtgtggga taagggccgg gactttgcca ccgtgcggaa agtgctgtct
     7261 atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag
     7321 tctatcctgc ccaagaggaa cagcgacaag ctgatcgcca gaaagaagga ctgggaccct
     7381 aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa
     7441 gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc
     7501 atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa
     7561 gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac
     7621 ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg
     7681 ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc
     7741 cccgaggata atgagcagaa acagctgttt gtggaacagc acaaacacta cctggacgag
     7801 atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac
     7861 aaggtgctga gcgcctacaa caagcacaga gacaagccta tcagagagca ggccgagaat
     7921 atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac
     7981 accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc
     8041 caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac
     8101 gccgactata aggaccacga cggagactac aaggatcatg atattgatta caaagacgat
     8161 gacgataaga gcctgggcag cggctccccc aagaaaaaac gcaaggtgga agatcctaag
     8221 aaaaagcgga aagtgtaagc tagcagatct ttttccctct gccaaaaatt atggggacat
     8281 catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat
     8341 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     8401 aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc
     8461 catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca
     8521 ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt
     8581 gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt
     8641 cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga gatccctcga
     8701 cctgcagccc aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     8761 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     8821 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     8881 acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca accatagtcc cgcccctaac
     8941 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     9001 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     9061 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct
     9121 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     9181 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatccgc
     9241 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     9301 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     9361 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     9421 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     9481 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     9541 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     9601 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     9661 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     9721 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     9781 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     9841 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     9901 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     9961 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
    10021 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
    10081 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
    10141 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
    10201 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
    10261 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
    10321 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
    10381 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
    10441 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
    10501 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
    10561 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
    10621 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
    10681 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
    10741 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
    10801 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
    10861 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
    10921 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
    10981 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
    11041 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
    11101 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
    11161 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
//
