LOCUS       sequence-453884-       10678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  miniEDV production plasmid. Expresses the miniGag-Cas9 polypeptide 
            (CAG promoter) and sgRNA (human U6 promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10678)
  AUTHORS   Ngo W, Peukes J, Baldwin A, Xue ZW, Hwang S, Stickels RR, Lin Z, 
            Satpathy AT, Wells JA, Schekman R, Nogales E, Doudna JA
  TITLE     Mechanism-guided engineering of a minimal biological particle for 
            genome editing.
  JOURNAL   Proc Natl Acad Sci U S A. 2025 Jan 7;122(1):e2413519121. doi: 
            10.1073/pnas.2413519121. Epub 2024 Dec 30.
  PUBMED    39793042
REFERENCE   2  (bases 1 to 10678)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2025-01-7"; volume: "122(1):e2413519121. 
            doi"; pages: " 10.1073/pnas.2413519121. Epub 2024 Dec 30"
COMMENT     Sequence Label: pWN_U6-TRAC-miniGag-Cas9
FEATURES             Location/Qualifiers
     source          1..10678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             45..65
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             72..92
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             99..119
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             126..146
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(159..178)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    224..279
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(278..297)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(640..656)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(640..656)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(653..675)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    664..680
                     /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(688..694,695..712,713..718))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    733..754
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        813..1008
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      859..994
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1014..1148
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1051..1070)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1105..1124
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1216..1233)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1387..1975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1467..1486)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2146..2937,2938..3006))
                     /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     2769..2788
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3007..3111)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3193..3441
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3193..3213
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3364..3383
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        3594..3973
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3975..4251
                     /label=chicken beta-actin promoter
     intron          4252..5268
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     5192..5214
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     5276..5295
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      5323..5332
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6256..6282
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             6295..6321
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             6334..6360
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             6532..10635
                     /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             6532..10635
                     /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             join(10639..10678,1..26)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 ttgattacaa agacgatgac gataagagcc tgggcagcgg ctcccccaag aaaaagagaa
       61 aagtcgaaga ccccaagaag aaaaggaagg tcgaagatcc taaaaagaaa aggaaagtgg
      121 aagatcctaa gaaaaagcgg aaagtgtaag ctagcagatc tttttccctc tgccaaaaat
      181 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
      241 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
      301 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
      361 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
      421 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
      481 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
      541 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
      601 agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
      661 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
      721 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
      781 ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc
      841 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
      901 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
      961 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctaacttgt
     1021 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1081 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     1141 tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1201 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1261 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1321 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1381 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1441 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1501 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1561 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1621 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1681 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1741 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1801 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1861 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1921 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1981 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2041 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2101 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2161 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2221 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2281 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2341 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2401 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2461 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     2521 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2581 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2641 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     2701 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     2761 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     2821 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     2881 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     2941 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3001 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3061 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3121 ccgaaaagtg ccacctggtc gacgggttta ttacagggac agcagagatc cagtttggtt
     3181 aattaaggta ccgagggcct atttcccatg attccttcat atttgcatat acgatacaag
     3241 gctgttagag agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat
     3301 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
     3361 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
     3421 ttgtggaaag gacgaaacac cgcagggttc tggatatctg tgtttaagag ctatgctgga
     3481 aacagcatag caagtttaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
     3541 tcggtgcttt ttttgaattc gctagctagg tcttgaaagg agtgggaatt gtcgacattg
     3601 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     3661 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     3721 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     3781 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     3841 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     3901 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     3961 cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc
     4021 ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc
     4081 gggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg
     4141 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
     4201 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct
     4261 gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
     4321 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
     4381 aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag
     4441 gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg
     4501 tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc
     4561 ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc
     4621 ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg
     4681 gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc
     4741 cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg
     4801 ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct
     4861 cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga
     4921 ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt
     4981 cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc
     5041 gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
     5101 gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac
     5161 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc
     5221 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
     5281 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcgccaccat gggtgcgaga
     5341 gcgtcagtat taagcggggg agaattagat cgatgggaaa aaattcggtt aaggccaggg
     5401 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
     5461 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
     5521 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
     5581 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
     5641 gaggaagagc aaaacaaaag taagaaaaaa gcacagcaag cagcagctga cacaggacac
     5701 agcaatcagg tcagccaaaa ttaccctata gtgcagagca ttctggacat aagacaagga
     5761 ccaaaggaac cctttagaga ctatgtagac cgattctata aaactctaag agccgagcaa
     5821 gcttcacaag aggtaaaaaa ttggatgaca gaaaccttgt tggtccaaaa tgcgaaccca
     5881 gattgtaaga ctattttaaa agcattggga ccaggagcga cactagaaga aatgatgaca
     5941 gcatgtcagg gagtgggggg acccggccat aaagcaagag ttttggctga agcaatgagc
     6001 caagtaacaa atccagctac cataatgata cagaaaggca gacaggctaa ttttttaggg
     6061 aagatctggc cttcccacaa gggaaggcca gggaattttc ttcagagcag accagagcca
     6121 acagccccac cagaagagag cttcaggttt ggggaagaga caacaactcc ctctcagaag
     6181 caggagccga tagacaagga actgtatcct ttagcttccc tcagatcact ctttggcagc
     6241 gacccctcgt cacaactccc ccctctggag cgccttactc ttggctccct gcagttgccc
     6301 ccactggagc ggctgaccct ggggtcattg cagttgcctc cactcgaacg actgacgttg
     6361 agccagaact atccgattgt gcagaccggt atgaagcgag cactgcccaa caacaccagc
     6421 tcctctcccc agccaaagaa gaaaccagga gggtcagggg gaaagcgagc actgcccaac
     6481 aacaccagct cctctcccca gccaaagaag aaaccaggag ggtcaggggg aatggacaag
     6541 aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac
     6601 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
     6661 aagaagaacc tgatcggcgc cctgctgttc gacagcggag aaacagccga ggccacccgg
     6721 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
     6781 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
     6841 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
     6901 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
     6961 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
     7021 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
     7081 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
     7141 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa
     7201 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggcaa cctgattgcc
     7261 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
     7321 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
     7381 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
     7441 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
     7501 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
     7561 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatc
     7621 gatggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
     7681 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg
     7741 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
     7801 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
     7861 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
     7921 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
     7981 aagggcgcca gcgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
     8041 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtacaacgag
     8101 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
     8161 cagaaaaaag ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
     8221 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
     8281 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
     8341 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
     8401 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
     8461 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
     8521 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
     8581 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
     8641 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
     8701 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
     8761 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
     8821 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
     8881 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
     8941 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
     9001 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct
     9061 cagagctttc tgaaggacga ctccatcgat aacaaagtgc tgactcggag cgacaagaac
     9121 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
     9181 cgccagctgc tgaatgccaa gctgattacc cagaggaagt tcgacaatct gaccaaggcc
     9241 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
     9301 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
     9361 gacgagaacg acaaactgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
     9421 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
     9481 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
     9541 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
     9601 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
     9661 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
     9721 gagacaaacg gcgaaacagg cgagatcgtg tgggataagg gccgggactt tgccaccgtg
     9781 cggaaagtgc tgtctatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
     9841 ggcttcagca aagagtctat cctgcccaag aggaacagcg acaagctgat cgccagaaag
     9901 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
     9961 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
    10021 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
    10081 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
    10141 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga
    10201 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
    10261 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaaa
    10321 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
    10381 gacgctaatc tggacaaggt gctgagcgcc tacaacaagc acagagacaa gcctatcaga
    10441 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
    10501 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
    10561 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct
    10621 cagctgggag gcgacgccga ctataaggac cacgacggag actacaagga tcatgata
//
