LOCUS       sequence-382813-       15559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dox-inducible all-in-one lentiviral construct to express 
            dCpf1-CTCF-Q418A mutant and crRNA-1 and -2 targeting the human MECP2
            locus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15559)
  AUTHORS   Qian J, Guan X, Xie B, Xu C, Niu J, Tang X, Li CH, Colecraft HM, 
            Jaenisch R, Liu XS
  TITLE     Multiplex epigenome editing of MECP2 to rescue Rett syndrome 
            neurons.
  JOURNAL   Sci Transl Med. 2023 Jan 18;15(679):eadd4666. doi: 
            10.1126/scitranslmed.add4666. Epub 2023 Jan 18.
  PUBMED    36652535
REFERENCE   2  (bases 1 to 15559)
  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: "Sci Transl 
            Med."; date: "2023-01-18"; volume: "15(679):eadd4666. doi"; pages: "
            10.1126/scitranslmed.add4666. Epub 2023 Jan 18"
COMMENT     Sequence Label: 
            FUW-crRNA1and2-U6_TRE-dLbCpf1-NLS-hCTCF_Q418A-HA-P2A-tagBFP
FEATURES             Location/Qualifiers
     source          1..15559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        complement(2703..2943)
                     /label=U6 promoter
                     /note="Truncated RNA polymerase III promoter for human U6 
                     snRNA used in pSIREN vectors such as 
                     pSIREN-RetroQ-ZsGreen1. Expected transcription start site 
                     is 8bp downstream of this feature."
     primer_bind     complement(2753..2772)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(2923..2943)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        3156..3534
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    3164..3182
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3200..3218
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3236..3254
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3272..3290
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3308..3326
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3344..3362
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3380..3398
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3443..3467
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3564..7247
                     /codon_start=1
                     /product="CRISPR-associated protein from Lachnospiraceae 
                     bacterium ND2006 (Zetsche et al., 2015)"
                     /label=LbCpf1
                     /translation="MSKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAE
                     DYKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKE
                     IAKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFDNRENMFS
                     EEAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFEG
                     EFFNFVLTQEGIDVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVL
                     SDRESLSFYGEGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKN
                     GPAISTISKDIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLEQ
                     LQEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKD
                     LLDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYS
                     KDKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVNG
                     NYEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKL
                     IDFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLV
                     EEGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLKK
                     EELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKI
                     NTEVRVLLKHDDNPYVIGIARGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTD
                     YHSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNSG
                     FKNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQ
                     NGFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALD
                     YKNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQ
                     QGDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSRN
                     YEAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTS
                     VKH"
     CDS             7269..7289
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7293..7313
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9519..9545
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             9546..9602
                     /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             9603..10304
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    10323..10911
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10376..10396)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10794..10805)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10914..10930)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10915..10931)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             11436..11616
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11642..11659)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11648..11872
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11918..12346
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12005..12024)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12215..12236
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12355..12375)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12399..12609
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        12421..12617
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      12468..12603
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12530..12549
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12665..12712
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12731..13105
                     /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    13235..13356
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13272..13291)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13326..13345
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13405..13421)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13405..13421)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13418..13440)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13429..13445
                     /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(13453..13459,13460..13477,13478..13483))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13498..13519
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13636..13653)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13807..14395)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13887..13906)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14566..15357,15358..15426))
                     /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     15189..15208
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15427..15531)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta aaaaaatgag cgggcctggg ccatgtccta tctacactta
     2641 gtagaaattg acccccttca catagagaga agatctacac ttagtagaaa ttgacgctag
     2701 ccgtcctttc cacaagatat ataaagccaa gaaatcgaaa tactttcaag ttacggtaag
     2761 catatgatag tccattttaa aacataattt taaaactgca aactacccaa gaaattatta
     2821 ctttctacgt cacgtatttt gtactaatat ctttgtgttt acagtcaaat taattctaat
     2881 tatctctcta acagccttgt atcgtatatg caaatatgaa ggaatcatgg gaaataggcc
     2941 ctcttcctgc ccgaccttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     3001 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctggt tttagagcta
     3061 gaaatagcaa gttaaaataa ggctagtccg tttttagcgc gtgcgccaat tctgcagaca
     3121 aatggctcta gattcgaagt cttcaagaat tgctcgagtt tactccctat cagtgataga
     3181 gaacgtatga agagtttact ccctatcagt gatagagaac gtatgcagac tttactccct
     3241 atcagtgata gagaacgtat aaggagttta ctccctatca gtgatagaga acgtatgacc
     3301 agtttactcc ctatcagtga tagagaacgt atctacagtt tactccctat cagtgataga
     3361 gaacgtatat ccagtttact ccctatcagt gatagagaac gtataagctt taggcgtgta
     3421 cggtgggcgc ctataaaagc agagctcgtt tagtgaaccg tcagatcgcc tggagcaatt
     3481 ccacaacact tttgtcttat accaactttc cgtaccactt cctaccctcg taaaccgcgg
     3541 gcccgggttc caccggtgcc accatgagca agctggagaa gtttacaaac tgctactccc
     3601 tgtctaagac cctgaggttc aaggccatcc ctgtgggcaa gacccaggag aacatcgaca
     3661 ataagcggct gctggtggag gacgagaaga gagccgagga ttataagggc gtgaagaagc
     3721 tgctggatcg ctactatctg tcttttatca acgacgtgct gcacagcatc aagctgaaga
     3781 atctgaacaa ttacatcagc ctgttccgga agaaaaccag aaccgagaag gagaataagg
     3841 agctggagaa cctggagatc aatctgcgga aggagatcgc caaggccttc aagggcaacg
     3901 agggctacaa gtccctgttt aagaaggata tcatcgagac aatcctgcca gagttcctgg
     3961 acgataagga cgagatcgcc ctggtgaaca gcttcaatgg ctttaccaca gccttcaccg
     4021 gcttctttga taacagagag aatatgtttt ccgaggaggc caagagcaca tccatcgcct
     4081 tcaggtgtat caacgagaat ctgacccgct acatctctaa tatggacatc ttcgagaagg
     4141 tggacgccat ctttgataag cacgaggtgc aggagatcaa ggagaagatc ctgaacagcg
     4201 actatgatgt ggaggatttc tttgagggcg agttctttaa ctttgtgctg acacaggagg
     4261 gcatcgacgt gtataacgcc atcatcggcg gcttcgtgac cgagagcggc gagaagatca
     4321 agggcctgaa cgagtacatc aacctgtata atcagaaaac caagcagaag ctgcctaagt
     4381 ttaagccact gtataagcag gtgctgagcg atcgggagtc tctgagcttc tacggcgagg
     4441 gctatacatc cgatgaggag gtgctggagg tgtttagaaa caccctgaac aagaacagcg
     4501 agatcttcag ctccatcaag aagctggaga agctgttcaa gaattttgac gagtactcta
     4561 gcgccggcat ctttgtgaag aacggccccg ccatcagcac aatctccaag gatatcttcg
     4621 gcgagtggaa cgtgatccgg gacaagtgga atgccgagta tgacgatatc cacctgaaga
     4681 agaaggccgt ggtgaccgag aagtacgagg acgatcggag aaagtccttc aagaagatcg
     4741 gctccttttc tctggagcag ctgcaggagt acgccgacgc cgatctgtct gtggtggaga
     4801 agctgaagga gatcatcatc cagaaggtgg atgagatcta caaggtgtat ggctcctctg
     4861 agaagctgtt cgacgccgat tttgtgctgg agaagagcct gaagaagaac gacgccgtgg
     4921 tggccatcat gaaggacctg ctggattctg tgaagagctt cgagaattac atcaaggcct
     4981 tctttggcga gggcaaggag acaaacaggg acgagtcctt ctatggcgat tttgtgctgg
     5041 cctacgacat cctgctgaag gtggaccaca tctacgatgc catccgcaat tatgtgaccc
     5101 agaagcccta ctctaaggat aagttcaagc tgtattttca gaaccctcag ttcatgggcg
     5161 gctgggacaa ggataaggag acagactatc gggccaccat cctgagatac ggctccaagt
     5221 actatctggc catcatggat aagaagtacg ccaagtgcct gcagaagatc gacaaggacg
     5281 atgtgaacgg caattacgag aagatcaact ataagctgct gcccggccct aataagatgc
     5341 tgccaaaggt gttcttttct aagaagtgga tggcctacta taaccccagc gaggacatcc
     5401 agaagatcta caagaatggc acattcaaga agggcgatat gtttaacctg aatgactgtc
     5461 acaagctgat cgacttcttt aaggatagca tctcccggta tccaaagtgg tccaatgcct
     5521 acgatttcaa cttttctgag acagagaagt ataaggacat cgccggcttt tacagagagg
     5581 tggaggagca gggctataag gtgagcttcg agtctgccag caagaaggag gtggataagc
     5641 tggtggagga gggcaagctg tatatgttcc agatctataa caaggacttt tccgataagt
     5701 ctcacggcac acccaatctg cacaccatgt acttcaagct gctgtttgac gagaacaatc
     5761 acggacagat caggctgagc ggaggagcag agctgttcat gaggcgcgcc tccctgaaga
     5821 aggaggagct ggtggtgcac ccagccaact cccctatcgc caacaagaat ccagataatc
     5881 ccaagaaaac cacaaccctg tcctacgacg tgtataagga taagaggttt tctgaggacc
     5941 agtacgagct gcacatccca atcgccatca ataagtgccc caagaacatc ttcaagatca
     6001 atacagaggt gcgcgtgctg ctgaagcacg acgataaccc ctatgtgatc ggcatcgcga
     6061 ggggcgagcg caatctgctg tatatcgtgg tggtggacgg caagggcaac atcgtggagc
     6121 agtattccct gaacgagatc atcaacaact tcaacggcat caggatcaag acagattacc
     6181 actctctgct ggacaagaag gagaaggaga ggttcgaggc ccgccagaac tggacctcca
     6241 tcgagaatat caaggagctg aaggccggct atatctctca ggtggtgcac aagatctgcg
     6301 agctggtgga gaagtacgat gccgtgatcg ccctggagga cctgaactct ggctttaaga
     6361 atagccgcgt gaaggtggag aagcaggtgt atcagaagtt cgagaagatg ctgatcgata
     6421 agctgaacta catggtggac aagaagtcta atccttgtgc aacaggcggc gccctgaagg
     6481 gctatcagat caccaataag ttcgagagct ttaagtccat gtctacccag aacggcttca
     6541 tcttttacat ccctgcctgg ctgacatcca agatcgatcc atctaccggc tttgtgaacc
     6601 tgctgaaaac caagtatacc agcatcgccg attccaagaa gttcatcagc tcctttgaca
     6661 ggatcatgta cgtgcccgag gaggatctgt tcgagtttgc cctggactat aagaacttct
     6721 ctcgcacaga cgccgattac atcaagaagt ggaagctgta ctcctacggc aaccggatca
     6781 gaatcttccg gaatcctaag aagaacaacg tgttcgactg ggaggaggtg tgcctgacca
     6841 gcgcctataa ggagctgttc aacaagtacg gcatcaatta tcagcagggc gatatcagag
     6901 ccctgctgtg cgagcagtcc gacaaggcct tctactctag ctttatggcc ctgatgagcc
     6961 tgatgctgca gatgcggaac agcatcacag gccgcaccga cgtggatttt ctgatcagcc
     7021 ctgtgaagaa ctccgacggc atcttctacg atagccggaa ctatgaggcc caggagaatg
     7081 ccatcctgcc aaagaacgcc gacgccaatg gcgcctataa catcgccaga aaggtgctgt
     7141 gggccatcgg ccagttcaag aaggccgagg acgagaagct ggataaggtg aagatcgcca
     7201 tctctaacaa ggagtggctg gagtacgccc agaccagcgt gaagcacgaa ttcggtggcg
     7261 gaggctcgcc aaaaaagaag agaaaggtag gtccaaagaa aaaacgaaaa gtagatggtg
     7321 gcggaggatc cgaaggtgat gcagtcgaag ccattgtgga ggagtccgaa acttttatta
     7381 aaggaaagga gagaaagact taccagagac gccgggaagg gggccaggaa gaagatgcct
     7441 gccacttacc ccagaaccag acggatgggg gtgaggtggt ccaggatgtc aacagcagtg
     7501 tacagatggt gatgatggaa cagctggacc ccacccttct tcagatgaag actgaagtaa
     7561 tggagggcac agtggctcca gaagcagagg ctgctgtgga cgatacccag attataactt
     7621 tacaggttgt aaatatggag gaacagccca taaacatagg agaacttcag cttgttcaag
     7681 tacctgttcc tgtgactgta cctgttgcta ccacttcagt agaagaactt cagggggctt
     7741 atgaaaatga agtgtctaaa gagggccttg cggaaagtga acccatgata tgccacaccc
     7801 tacctttgcc tgaagggttt caggtggtta aagtgggggc caatggagag gtggagacac
     7861 tagaacaagg ggaacttcca ccccaggaag atcctagttg gcaaaaagac ccagactatc
     7921 agccaccagc caaaaaaaca aagaaaacca aaaagagcaa actgcgttat acagaggagg
     7981 gcaaagatgt agatgtgtct gtctacgatt ttgaggaaga acagcaggag ggtctgctat
     8041 cagaggttaa tgcagagaaa gtggttggta atatgaagcc tccaaagcca acaaaaatta
     8101 aaaagaaagg tgtaaagaag acattccagt gtgagctttg cagttacacg tgtccacggc
     8161 gttcaaattt ggatcgtcac atgaaaagcc acactgatga gagaccacac aagtgccatc
     8221 tctgtggcag ggcattcaga acagtcaccc tcctgaggaa tcaccttaac acacacacag
     8281 gtactcgtcc tcacaagtgc ccagactgcg acatggcctt tgtgaccagt ggagaattgg
     8341 ttcggcatcg tcgttacaaa cacacccacg agaagccatt caagtgttcc atgtgcgatt
     8401 acgccagtgt agaagtcagc aaattaaaac gtcacattcg ctctcatact ggagagcgtc
     8461 cgtttcagtg cagtttgtgc agttatgcca gcagggacac atacaagctg aaaaggcaca
     8521 tgagaaccca ttcaggggaa aagccttatg aatgttatat ttgtcatgct cggtttaccg
     8581 caagtggtac catgaagatg cacattttac agaagcacac agaaaatgtg gccaaatttc
     8641 actgtcccca ctgtgacaca gtcatagccc gaaaaagtga tttgggtgtc cacttgcgaa
     8701 agcagcattc ctatattgag caaggcaaga aatgccgtta ctgtgatgct gtgtttcatg
     8761 agcgctatgc cctcatccag catcagaagt cacacaagaa tgagaagcgc tttaagtgtg
     8821 accagtgtga ttacgcttgt agacaggaga ggcacatgat catgcacaag cgcacccaca
     8881 ccggggagaa gccttacgcc tgcagccact gcgataagac cttccgccag aagcagcttc
     8941 tcgacatgca cttcaagcgc tatcacgacc ccaacttcgt ccctgcggct tttgtctgtt
     9001 ctaagtgtgg gaaaacattt acacgtcgga ataccatggc aagacatgct gataattgtg
     9061 ctggcccaga tggcgtagag ggggaaaatg gaggagaaac gaagaagagt aaacgtggaa
     9121 gaaaaagaaa gatgcgctct aagaaagaag attcctctga cagtgaaaat gctgaaccag
     9181 atctggacga caatgaggat gaggaggagc ctgccgtaga aattgaacct gagccagagc
     9241 ctcagcctgt gaccccagcc ccaccacccg ccaagaagcg gagaggacga ccccctggca
     9301 gaaccaacca gcccaaacag aaccagccaa cagctatcat tcaggttgaa gaccagaata
     9361 caggtgcaat tgagaacatt atagttgaag taaaaaaaga gccagatgct gagcccgcag
     9421 agggagagga agaggaggcc cagccagctg ccacagatgc ccccaacgga gacctcacgc
     9481 ccgagatgat cctcagcatg atggaccggg ttaacgcata cccctacgac gtccccgact
     9541 acgccgccac gaacttctct ctgttaaagc aagcaggaga tgttgaagaa aaccccgggc
     9601 ctatgagcga gctgattaag gagaacatgc acatgaagct gtacatggag ggcaccgtgg
     9661 acaaccatca cttcaagtgc acatccgagg gcgaaggcaa gccctacgag ggcacccaga
     9721 ccatgagaat caaggtggtc gagggcggcc ctctcccctt cgccttcgac atcctggcta
     9781 ctagcttcct ctacggcagc aagaccttca tcaaccacac ccagggcatc cccgacttct
     9841 tcaagcagtc cttccctgag ggcttcacat gggagagagt caccacatac gaagacgggg
     9901 gcgtgctgac cgctacccag gacaccagcc tccaggacgg ctgcctcatc tacaacgtca
     9961 agatcagagg ggtgaacttc acatccaacg gccctgtgat gcagaagaaa acactcggct
    10021 gggaggcctt caccgagacg ctgtaccccg ctgacggcgg cctggaaggc agaaacgaca
    10081 tggccctgaa gctcgtgggc gggagccatc tgatcgcaaa catcaagacc acatatagat
    10141 ccaagaaacc cgctaagaac ctcaagatgc ctggcgtcta ctatgtggac tacagactgg
    10201 aaagaatcaa ggaggccaac aacgagacct acgtcgagca gcacgaggtg gcagtggcca
    10261 gatactgcga cctccctagc aaactggggc acaagcttaa ttgagatatc aagcttatcg
    10321 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
    10381 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
    10441 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
    10501 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
    10561 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
    10621 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
    10681 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
    10741 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
    10801 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
    10861 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc
    10921 gtcgacctcg agacctagaa aaacatggag caatcacaag tagcaataca gcagctacca
    10981 atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt ccagtcacac
    11041 ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
    11101 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat atccttgatc
    11161 tgtggatcta ccacacacaa ggctacttcc ctgattggca gaactacaca ccagggccag
    11221 ggatcagata tccactgacc tttggatggt gctacaagct agtaccagtt gagcaagaga
    11281 aggtagaaga agccaatgaa ggagagaaca cccgcttgtt acaccctgtg agcctgcatg
    11341 ggatggatga cccggagaga gaagtattag agtggaggtt tgacagccgc ctagcatttc
    11401 atcacatggc ccgagagctg catccggact gtactgggtc tctctggtta gaccagatct
    11461 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
    11521 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
    11581 tcagaccctt ttagtcagtg tggaaaatct ctagcagggc ccgtttaaac ccgctgatca
    11641 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
    11701 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
    11761 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
    11821 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
    11881 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
    11941 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
    12001 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
    12061 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
    12121 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
    12181 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
    12241 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
    12301 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
    12361 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
    12421 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
    12481 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
    12541 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
    12601 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
    12661 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
    12721 gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc
    12781 cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga
    12841 cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt
    12901 ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga
    12961 gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat
    13021 cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca
    13081 cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt
    13141 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
    13201 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
    13261 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
    13321 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
    13381 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
    13441 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
    13501 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
    13561 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
    13621 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
    13681 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
    13741 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
    13801 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
    13861 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
    13921 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
    13981 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
    14041 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
    14101 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
    14161 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
    14221 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
    14281 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
    14341 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
    14401 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
    14461 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
    14521 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
    14581 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
    14641 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
    14701 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
    14761 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
    14821 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
    14881 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
    14941 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
    15001 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
    15061 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
    15121 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
    15181 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
    15241 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
    15301 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
    15361 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
    15421 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
    15481 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
    15541 ccgaaaagtg ccacctgac
//
