LOCUS       sequence-392583-       15516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dox-inducible all-in-one lentiviral construct to express 
            dCpf1-CTCF-K365A_R368A mutant and crRNA-1 targeting the human MECP2 
            locus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15516)
  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 15516)
  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-crRNA-1-U6_TRE-dLbCpf1-NLS-hCTCF_K365A_R368A-HA-P2A-tagBFP
FEATURES             Location/Qualifiers
     source          1..15516
                     /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(2660..2900)
                     /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(2710..2729)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(2880..2900)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        3113..3491
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    3121..3139
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3157..3175
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3193..3211
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3229..3247
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3265..3283
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3301..3319
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3337..3355
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3400..3424
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3521..7204
                     /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             7226..7246
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7250..7270
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9476..9502
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             9503..9559
                     /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             9560..10261
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    10280..10868
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10333..10353)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10751..10762)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10871..10887)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10872..10888)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             11393..11573
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11599..11616)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11605..11829
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11875..12303
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11962..11981)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12172..12193
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12312..12332)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12356..12566
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        12378..12574
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      12425..12560
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12487..12506
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12622..12669
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12688..13062
                     /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    13192..13313
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13229..13248)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13283..13302
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13362..13378)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13362..13378)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13375..13397)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13386..13402
                     /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(13410..13416,13417..13434,13435..13440))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13455..13476
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13593..13610)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13764..14352)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13844..13863)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14523..15314,15315..15383))
                     /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     15146..15165
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15384..15488)
                     /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 acgctagccg tcctttccac aagatatata aagccaagaa atcgaaatac
     2701 tttcaagtta cggtaagcat atgatagtcc attttaaaac ataattttaa aactgcaaac
     2761 tacccaagaa attattactt tctacgtcac gtattttgta ctaatatctt tgtgtttaca
     2821 gtcaaattaa ttctaattat ctctctaaca gccttgtatc gtatatgcaa atatgaagga
     2881 atcatgggaa ataggccctc ttcctgcccg accttattgt ctcatgagcg gatacatatt
     2941 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3001 acctggtttt agagctagaa atagcaagtt aaaataaggc tagtccgttt ttagcgcgtg
     3061 cgccaattct gcagacaaat ggctctagat tcgaagtctt caagaattgc tcgagtttac
     3121 tccctatcag tgatagagaa cgtatgaaga gtttactccc tatcagtgat agagaacgta
     3181 tgcagacttt actccctatc agtgatagag aacgtataag gagtttactc cctatcagtg
     3241 atagagaacg tatgaccagt ttactcccta tcagtgatag agaacgtatc tacagtttac
     3301 tccctatcag tgatagagaa cgtatatcca gtttactccc tatcagtgat agagaacgta
     3361 taagctttag gcgtgtacgg tgggcgccta taaaagcaga gctcgtttag tgaaccgtca
     3421 gatcgcctgg agcaattcca caacactttt gtcttatacc aactttccgt accacttcct
     3481 accctcgtaa accgcgggcc cgggttccac cggtgccacc atgagcaagc tggagaagtt
     3541 tacaaactgc tactccctgt ctaagaccct gaggttcaag gccatccctg tgggcaagac
     3601 ccaggagaac atcgacaata agcggctgct ggtggaggac gagaagagag ccgaggatta
     3661 taagggcgtg aagaagctgc tggatcgcta ctatctgtct tttatcaacg acgtgctgca
     3721 cagcatcaag ctgaagaatc tgaacaatta catcagcctg ttccggaaga aaaccagaac
     3781 cgagaaggag aataaggagc tggagaacct ggagatcaat ctgcggaagg agatcgccaa
     3841 ggccttcaag ggcaacgagg gctacaagtc cctgtttaag aaggatatca tcgagacaat
     3901 cctgccagag ttcctggacg ataaggacga gatcgccctg gtgaacagct tcaatggctt
     3961 taccacagcc ttcaccggct tctttgataa cagagagaat atgttttccg aggaggccaa
     4021 gagcacatcc atcgccttca ggtgtatcaa cgagaatctg acccgctaca tctctaatat
     4081 ggacatcttc gagaaggtgg acgccatctt tgataagcac gaggtgcagg agatcaagga
     4141 gaagatcctg aacagcgact atgatgtgga ggatttcttt gagggcgagt tctttaactt
     4201 tgtgctgaca caggagggca tcgacgtgta taacgccatc atcggcggct tcgtgaccga
     4261 gagcggcgag aagatcaagg gcctgaacga gtacatcaac ctgtataatc agaaaaccaa
     4321 gcagaagctg cctaagttta agccactgta taagcaggtg ctgagcgatc gggagtctct
     4381 gagcttctac ggcgagggct atacatccga tgaggaggtg ctggaggtgt ttagaaacac
     4441 cctgaacaag aacagcgaga tcttcagctc catcaagaag ctggagaagc tgttcaagaa
     4501 ttttgacgag tactctagcg ccggcatctt tgtgaagaac ggccccgcca tcagcacaat
     4561 ctccaaggat atcttcggcg agtggaacgt gatccgggac aagtggaatg ccgagtatga
     4621 cgatatccac ctgaagaaga aggccgtggt gaccgagaag tacgaggacg atcggagaaa
     4681 gtccttcaag aagatcggct ccttttctct ggagcagctg caggagtacg ccgacgccga
     4741 tctgtctgtg gtggagaagc tgaaggagat catcatccag aaggtggatg agatctacaa
     4801 ggtgtatggc tcctctgaga agctgttcga cgccgatttt gtgctggaga agagcctgaa
     4861 gaagaacgac gccgtggtgg ccatcatgaa ggacctgctg gattctgtga agagcttcga
     4921 gaattacatc aaggccttct ttggcgaggg caaggagaca aacagggacg agtccttcta
     4981 tggcgatttt gtgctggcct acgacatcct gctgaaggtg gaccacatct acgatgccat
     5041 ccgcaattat gtgacccaga agccctactc taaggataag ttcaagctgt attttcagaa
     5101 ccctcagttc atgggcggct gggacaagga taaggagaca gactatcggg ccaccatcct
     5161 gagatacggc tccaagtact atctggccat catggataag aagtacgcca agtgcctgca
     5221 gaagatcgac aaggacgatg tgaacggcaa ttacgagaag atcaactata agctgctgcc
     5281 cggccctaat aagatgctgc caaaggtgtt cttttctaag aagtggatgg cctactataa
     5341 ccccagcgag gacatccaga agatctacaa gaatggcaca ttcaagaagg gcgatatgtt
     5401 taacctgaat gactgtcaca agctgatcga cttctttaag gatagcatct cccggtatcc
     5461 aaagtggtcc aatgcctacg atttcaactt ttctgagaca gagaagtata aggacatcgc
     5521 cggcttttac agagaggtgg aggagcaggg ctataaggtg agcttcgagt ctgccagcaa
     5581 gaaggaggtg gataagctgg tggaggaggg caagctgtat atgttccaga tctataacaa
     5641 ggacttttcc gataagtctc acggcacacc caatctgcac accatgtact tcaagctgct
     5701 gtttgacgag aacaatcacg gacagatcag gctgagcgga ggagcagagc tgttcatgag
     5761 gcgcgcctcc ctgaagaagg aggagctggt ggtgcaccca gccaactccc ctatcgccaa
     5821 caagaatcca gataatccca agaaaaccac aaccctgtcc tacgacgtgt ataaggataa
     5881 gaggttttct gaggaccagt acgagctgca catcccaatc gccatcaata agtgccccaa
     5941 gaacatcttc aagatcaata cagaggtgcg cgtgctgctg aagcacgacg ataaccccta
     6001 tgtgatcggc atcgcgaggg gcgagcgcaa tctgctgtat atcgtggtgg tggacggcaa
     6061 gggcaacatc gtggagcagt attccctgaa cgagatcatc aacaacttca acggcatcag
     6121 gatcaagaca gattaccact ctctgctgga caagaaggag aaggagaggt tcgaggcccg
     6181 ccagaactgg acctccatcg agaatatcaa ggagctgaag gccggctata tctctcaggt
     6241 ggtgcacaag atctgcgagc tggtggagaa gtacgatgcc gtgatcgccc tggaggacct
     6301 gaactctggc tttaagaata gccgcgtgaa ggtggagaag caggtgtatc agaagttcga
     6361 gaagatgctg atcgataagc tgaactacat ggtggacaag aagtctaatc cttgtgcaac
     6421 aggcggcgcc ctgaagggct atcagatcac caataagttc gagagcttta agtccatgtc
     6481 tacccagaac ggcttcatct tttacatccc tgcctggctg acatccaaga tcgatccatc
     6541 taccggcttt gtgaacctgc tgaaaaccaa gtataccagc atcgccgatt ccaagaagtt
     6601 catcagctcc tttgacagga tcatgtacgt gcccgaggag gatctgttcg agtttgccct
     6661 ggactataag aacttctctc gcacagacgc cgattacatc aagaagtgga agctgtactc
     6721 ctacggcaac cggatcagaa tcttccggaa tcctaagaag aacaacgtgt tcgactggga
     6781 ggaggtgtgc ctgaccagcg cctataagga gctgttcaac aagtacggca tcaattatca
     6841 gcagggcgat atcagagccc tgctgtgcga gcagtccgac aaggccttct actctagctt
     6901 tatggccctg atgagcctga tgctgcagat gcggaacagc atcacaggcc gcaccgacgt
     6961 ggattttctg atcagccctg tgaagaactc cgacggcatc ttctacgata gccggaacta
     7021 tgaggcccag gagaatgcca tcctgccaaa gaacgccgac gccaatggcg cctataacat
     7081 cgccagaaag gtgctgtggg ccatcggcca gttcaagaag gccgaggacg agaagctgga
     7141 taaggtgaag atcgccatct ctaacaagga gtggctggag tacgcccaga ccagcgtgaa
     7201 gcacgaattc ggtggcggag gctcgccaaa aaagaagaga aaggtaggtc caaagaaaaa
     7261 acgaaaagta gatggtggcg gaggatccga aggtgatgca gtcgaagcca ttgtggagga
     7321 gtccgaaact tttattaaag gaaaggagag aaagacttac cagagacgcc gggaaggggg
     7381 ccaggaagaa gatgcctgcc acttacccca gaaccagacg gatgggggtg aggtggtcca
     7441 ggatgtcaac agcagtgtac agatggtgat gatggaacag ctggacccca cccttcttca
     7501 gatgaagact gaagtaatgg agggcacagt ggctccagaa gcagaggctg ctgtggacga
     7561 tacccagatt ataactttac aggttgtaaa tatggaggaa cagcccataa acataggaga
     7621 acttcagctt gttcaagtac ctgttcctgt gactgtacct gttgctacca cttcagtaga
     7681 agaacttcag ggggcttatg aaaatgaagt gtctaaagag ggccttgcgg aaagtgaacc
     7741 catgatatgc cacaccctac ctttgcctga agggtttcag gtggttaaag tgggggccaa
     7801 tggagaggtg gagacactag aacaagggga acttccaccc caggaagatc ctagttggca
     7861 aaaagaccca gactatcagc caccagccaa aaaaacaaag aaaaccaaaa agagcaaact
     7921 gcgttataca gaggagggca aagatgtaga tgtgtctgtc tacgattttg aggaagaaca
     7981 gcaggagggt ctgctatcag aggttaatgc agagaaagtg gttggtaata tgaagcctcc
     8041 aaagccaaca aaaattaaaa agaaaggtgt aaagaagaca ttccagtgtg agctttgcag
     8101 ttacacgtgt ccacggcgtt caaatttgga tcgtcacatg aaaagccaca ctgatgagag
     8161 accacacaag tgccatctct gtggcagggc attcagaaca gtcaccctcc tgaggaatca
     8221 ccttaacaca cacacaggta ctcgtcctca caagtgccca gactgcgaca tggcctttgt
     8281 gaccagtgga gaattggttc ggcatcgtcg ttacaaacac acccacgaga agccattcaa
     8341 gtgttccatg tgcgattacg ccagtgtaga agtcagcgca ttaaaagctc acattcgctc
     8401 tcatactgga gagcgtccgt ttcagtgcag tttgtgcagt tatgccagca gggacacata
     8461 caagctgaaa aggcacatga gaacccattc aggggaaaag ccttatgaat gttatatttg
     8521 tcatgctcgg tttacccaaa gtggtaccat gaagatgcac attttacaga agcacacaga
     8581 aaatgtggcc aaatttcact gtccccactg tgacacagtc atagcccgaa aaagtgattt
     8641 gggtgtccac ttgcgaaagc agcattccta tattgagcaa ggcaagaaat gccgttactg
     8701 tgatgctgtg tttcatgagc gctatgccct catccagcat cagaagtcac acaagaatga
     8761 gaagcgcttt aagtgtgacc agtgtgatta cgcttgtaga caggagaggc acatgatcat
     8821 gcacaagcgc acccacaccg gggagaagcc ttacgcctgc agccactgcg ataagacctt
     8881 ccgccagaag cagcttctcg acatgcactt caagcgctat cacgacccca acttcgtccc
     8941 tgcggctttt gtctgttcta agtgtgggaa aacatttaca cgtcggaata ccatggcaag
     9001 acatgctgat aattgtgctg gcccagatgg cgtagagggg gaaaatggag gagaaacgaa
     9061 gaagagtaaa cgtggaagaa aaagaaagat gcgctctaag aaagaagatt cctctgacag
     9121 tgaaaatgct gaaccagatc tggacgacaa tgaggatgag gaggagcctg ccgtagaaat
     9181 tgaacctgag ccagagcctc agcctgtgac cccagcccca ccacccgcca agaagcggag
     9241 aggacgaccc cctggcagaa ccaaccagcc caaacagaac cagccaacag ctatcattca
     9301 ggttgaagac cagaatacag gtgcaattga gaacattata gttgaagtaa aaaaagagcc
     9361 agatgctgag cccgcagagg gagaggaaga ggaggcccag ccagctgcca cagatgcccc
     9421 caacggagac ctcacgcccg agatgatcct cagcatgatg gaccgggtta acgcataccc
     9481 ctacgacgtc cccgactacg ccgccacgaa cttctctctg ttaaagcaag caggagatgt
     9541 tgaagaaaac cccgggccta tgagcgagct gattaaggag aacatgcaca tgaagctgta
     9601 catggagggc accgtggaca accatcactt caagtgcaca tccgagggcg aaggcaagcc
     9661 ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc
     9721 cttcgacatc ctggctacta gcttcctcta cggcagcaag accttcatca accacaccca
     9781 gggcatcccc gacttcttca agcagtcctt ccctgagggc ttcacatggg agagagtcac
     9841 cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg
     9901 cctcatctac aacgtcaaga tcagaggggt gaacttcaca tccaacggcc ctgtgatgca
     9961 gaagaaaaca ctcggctggg aggccttcac cgagacgctg taccccgctg acggcggcct
    10021 ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg agccatctga tcgcaaacat
    10081 caagaccaca tatagatcca agaaacccgc taagaacctc aagatgcctg gcgtctacta
    10141 tgtggactac agactggaaa gaatcaagga ggccaacaac gagacctacg tcgagcagca
    10201 cgaggtggca gtggccagat actgcgacct ccctagcaaa ctggggcaca agcttaattg
    10261 agatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
    10321 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
    10381 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
    10441 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
    10501 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
    10561 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
    10621 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
    10681 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
    10741 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
    10801 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
    10861 ctccccgcat cgataccgtc gacctcgaga cctagaaaaa catggagcaa tcacaagtag
    10921 caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt
    10981 gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga
    11041 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
    11101 acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg attggcagaa
    11161 ctacacacca gggccaggga tcagatatcc actgaccttt ggatggtgct acaagctagt
    11221 accagttgag caagagaagg tagaagaagc caatgaagga gagaacaccc gcttgttaca
    11281 ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtattagagt ggaggtttga
    11341 cagccgccta gcatttcatc acatggcccg agagctgcat ccggactgta ctgggtctct
    11401 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
    11461 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
    11521 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagggcccg
    11581 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
    11641 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
    11701 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
    11761 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
    11821 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
    11881 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
    11941 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
    12001 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
    12061 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
    12121 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
    12181 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
    12241 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
    12301 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
    12361 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
    12421 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
    12481 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
    12541 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
    12601 tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta
    12661 taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc
    12721 accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg
    12781 gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc
    12841 gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg
    12901 gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg
    12961 ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg
    13021 gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc
    13081 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
    13141 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
    13201 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
    13261 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
    13321 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
    13381 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
    13441 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
    13501 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
    13561 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
    13621 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
    13681 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
    13741 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
    13801 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
    13861 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
    13921 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
    13981 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
    14041 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
    14101 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
    14161 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
    14221 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
    14281 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
    14341 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
    14401 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
    14461 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
    14521 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
    14581 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
    14641 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
    14701 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
    14761 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
    14821 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
    14881 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
    14941 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
    15001 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
    15061 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
    15121 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
    15181 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
    15241 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
    15301 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
    15361 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
    15421 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
    15481 gttccgcgca catttccccg aaaagtgcca cctgac
//
