LOCUS       P3-Lenti-dCas9-T       16266 bp DNA     circular SYN 15-AUG-2022
DEFINITION  Lentiviral construct to express dCas9 fused with the catalytic 
            domain of Tet1.
ACCESSION   .
VERSION     .
KEYWORDS    P3-Lenti-dCas9-Tet1-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16266)
  AUTHORS   Liu XS, Wu H, Ji X, Stelzer Y, Wu X, Czauderna S, Shu J, Dadon D, 
            Young RA, Jaenisch R
  TITLE     Editing DNA Methylation in the Mammalian Genome.
  JOURNAL   Cell. 2016 Sep 22;167(1):233-247.e17. doi: 
            10.1016/j.cell.2016.08.056.
  PUBMED    27662091
REFERENCE   2  (bases 1 to 16266)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 15, 2022 from SnapGene 6.1.0
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2016.08"; journalName: "Cell"; date: "2016-09-22- 
            22"; volume: "167"; issue: "1"; pages: "233-247.e17"
FEATURES             Location/Qualifiers
     source          1..16266
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    12..33
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(150..167)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(321..909)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(401..420)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1080..1940)
                     /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     1703..1722
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1941..2045)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2120..2139)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2311..2690
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2692..2890
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2845..2865
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2908..3088
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3135..3260
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3753..3986
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    4513..4630
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4695..5906
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5743..5762
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5963..10066
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             10082..10102
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10106..10126
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10130..10150
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10157..12310
                     /codon_start=1
                     /label=Active Tet1
                     /translation="LPTCSCLDRVIQKDKGPYYTHLGAGPSVAAVREIMENRYGQKGNA
                     IRIEIVVYTGKEGKSSHGCPIAKWVLRRSSDEEKVLCLVRQRTGHHCPTAVMVVLIMVW
                     DGIPLPMADRLYTELTENLKSYNGHPTDRRCTLNENRTCTCQGIDPETCGASFSFGCSW
                     SMYFNGCKFGRSPSPRRFRIDPSSPLHEKNLEDNLQSLATRLAPIYKQYAPVAYQNQVE
                     YENVARECRLGSKEGRPFSGVTACLDFCAHPHRDIHNMNNGSTVVCTLTREDNRSLGVI
                     PQDEQLHVLPLYKLSDTDEFGSKEGMEAKIKSGAIEVLAPRRKKRTCFTQPVPRSGKKR
                     AAMMTEVLAHKIRAVEKKPIPRIKRKNNSTTTNNSKPSSLPTLGSNTETVQPEVKSETE
                     PHFILKSSDNTKTYSLMPSAPHPVKEASPGFSWSPKTASATPAPLKNDATASCGFSERS
                     STPHCTMPSGRLSGANAAAADGPGISQLGEVAPLPTLSAPVMEPLINSEPSTGVTEPLT
                     PHQPNHQPSFLTSPQDLASSPMEEDEQHSEADEPPSDEPLSDDPLSPAEEKLPHIDEYW
                     SDSEHIFLDANIGGVAIAPAHGSVLIECARRELHATTPVEHPNRNHPTRLSLVFYQHKN
                     LNKPQHGFELNKIKFEAKEAKNKKMKASEQKDQAANEGPEQSSEVNELNQIPSHKALTL
                     THDNVVTVSPYALTHVAGPYNHWV"
     primer_bind     11283..11301
                     /label=Tet1-F qPCR
     CDS             12311..12364
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             12365..13078
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(12407..12428)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(12668..12687)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     13015..13036
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    13103..13691
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(13156..13176)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(13574..13585)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(13694..13710)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13695..13711)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             14216..14396
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(14422..14439)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    14428..14652
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      14698..15126
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14785..14804)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14995..15016
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(15135..15155)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        15179..15389
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        15201..15397
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      15248..15383
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     15310..15329
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        15445..15492
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             15511..15885
                     /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    16015..16136
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(16052..16071)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     16106..16125
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(16185..16201)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(16185..16201)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(16198..16220)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    16209..16225
                     /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(16233..16263)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
       61 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
      121 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
      181 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
      241 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      301 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      361 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      421 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      481 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
      541 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      601 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      661 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      721 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      781 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      841 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      901 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
      961 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1021 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     1081 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1141 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     1201 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     1261 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     1321 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     1381 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     1441 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     1501 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     1561 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     1621 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     1681 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     1741 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     1801 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     1861 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     1921 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     1981 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     2041 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     2101 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     2161 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     2221 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     2281 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
     2341 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     2401 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     2461 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     2521 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     2581 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     2641 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     2701 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     2761 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     2821 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     2881 atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
     2941 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     3001 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     3061 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     3121 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     3181 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     3241 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     3301 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     3361 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     3421 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     3481 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     3541 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     3601 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     3661 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     3721 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     3781 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
     3841 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     3901 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     3961 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     4021 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
     4081 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
     4141 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
     4201 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
     4261 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
     4321 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
     4381 aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga
     4441 tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac aaatggcagt
     4501 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
     4561 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
     4621 tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttaa ttaacccgtg
     4681 tcggctccag atctggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcct
     4741 cacggcgagc gctgccacgt cagacgaagg gcgcagcgag cgtcctgatc cttccgcccg
     4801 gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca
     4861 gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag
     4921 cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg
     4981 cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca
     5041 gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag
     5101 cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg
     5161 tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg
     5221 ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt
     5281 gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa
     5341 ggtcttgagg ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctggggcac
     5401 catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt
     5461 gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc
     5521 gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc
     5581 tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt
     5641 aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc
     5701 agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa
     5761 ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt
     5821 aatcatttgg gtcaatatgt aattttcagt gttagactag taaattgtcc gctaaattct
     5881 ggccgttttt ggcttttttg ttagacgaag cttgggctgc aggtcgactc tagaggatcc
     5941 agttaacctc gagggcgcgc ccatggacaa gaagtattct atcggactgg ccatcgggac
     6001 taatagcgtc gggtgggccg tgatcactga cgagtacaag gtgccctcta agaagttcaa
     6061 ggtgctcggg aacaccgacc ggcattccat caagaaaaat ctgatcggag ctctcctctt
     6121 tgattcaggg gagaccgctg aagcaacccg cctcaagcgg actgctagac ggcggtacac
     6181 caggaggaag aaccggattt gttaccttca agagatattc tccaacgaaa tggcaaaggt
     6241 cgacgacagc ttcttccata ggctggaaga atcattcctc gtggaagagg ataagaagca
     6301 tgaacggcat cccatcttcg gtaatatcgt cgacgaggtg gcctatcacg agaaataccc
     6361 aaccatctac catcttcgca aaaagctggt ggactcaacc gacaaggcag acctccggct
     6421 tatctacctg gccctggccc acatgatcaa gttcagaggc cacttcctga tcgagggcga
     6481 cctcaatcct gacaatagcg atgtggataa actgttcatc cagctggtgc agacttacaa
     6541 ccagctcttt gaagagaacc ccatcaatgc aagcggagtc gatgccaagg ccattctgtc
     6601 agcccggctg tcaaagagcc gcagacttga gaatcttatc gctcagctgc cgggtgaaaa
     6661 gaaaaatgga ctgttcggga acctgattgc tctttcactt gggctgactc ccaatttcaa
     6721 gtctaatttc gacctggcag aggatgccaa gctgcaactg tccaaggaca cctatgatga
     6781 cgatctcgac aacctcctgg cccagatcgg tgaccaatac gccgaccttt tccttgctgc
     6841 taagaatctt tctgacgcca tcctgctgtc tgacattctc cgcgtgaaca ctgaaatcac
     6901 caaggcccct ctttcagctt caatgattaa gcggtatgat gagcaccacc aggacctgac
     6961 cctgcttaag gcactcgtcc ggcagcagct tccggagaag tacaaggaaa tcttctttga
     7021 ccagtcaaag aatggatacg ccggctacat cgacggaggt gcctcccaag aggaatttta
     7081 taagtttatc aaacctatcc ttgagaagat ggacggcacc gaagagctcc tcgtgaaact
     7141 gaatcgggag gatctgctgc ggaagcagcg cactttcgac aatgggagca ttccccacca
     7201 gatccatctt ggggagcttc acgccatcct tcggcgccaa gaggacttct acccctttct
     7261 taaggacaac agggagaaga ttgagaaaat tctcactttc cgcatcccct actacgtggg
     7321 acccctcgcc agaggaaata gccggtttgc ttggatgacc agaaagtcag aagaaactat
     7381 cactccctgg aacttcgaag aggtggtgga caagggagcc agcgctcagt cattcatcga
     7441 acggatgact aacttcgata agaacctccc caatgagaag gtcctgccga aacattccct
     7501 gctctacgag tactttaccg tgtacaacga gctgaccaag gtgaaatatg tcaccgaagg
     7561 gatgaggaag cccgcattcc tgtcaggcga acaaaagaag gcaattgtgg accttctgtt
     7621 caagaccaat agaaaggtga ccgtgaagca gctgaaggag gactatttca agaaaattga
     7681 atgcttcgac tctgtggaga ttagcggggt cgaagatcgg ttcaacgcaa gcctgggtac
     7741 ctaccatgat ctgcttaaga tcatcaagga caaggatttt ctggacaatg aggagaacga
     7801 ggacatcctt gaggacattg tcctgactct cactctgttc gaggaccggg aaatgatcga
     7861 ggagaggctt aagacctacg cccatctgtt cgacgataaa gtgatgaagc aacttaaacg
     7921 gagaagatat accggatggg gacgccttag ccgcaaactc atcaacggaa tccgggacaa
     7981 acagagcgga aagaccattc ttgatttcct taaaagcgac ggattcgcta atcgcaactt
     8041 catgcaactt atccatgatg attccctgac ctttaaggag gacatccaga aggcccaagt
     8101 gtctggacaa ggtgactcac tgcacgagca tatcgcaaat ctggctggtt cacccgctat
     8161 taagaagggt attctccaga ccgtgaaagt cgtggacgag ctggtcaagg tgatgggtcg
     8221 ccataaacca gagaacattg tcatcgagat ggccagggaa aaccagacta cccagaaggg
     8281 acagaagaac agcagggagc ggatgaaaag aattgaggaa gggattaagg agctcgggtc
     8341 acagatcctt aaagagcacc cggtggaaaa cacccagctt cagaatgaga agctctatct
     8401 gtactacctt caaaatggac gcgatatgta tgtggaccaa gagcttgata tcaacaggct
     8461 ctcagactac gacgtggacg ccatcgtccc tcagagcttc ctcaaagacg actcaattga
     8521 caataaggtg ctgactcgct cagacaagaa ccggggaaag tcagataacg tgccctcaga
     8581 ggaagtcgtg aaaaagatga agaactattg gcgccagctt ctgaacgcaa agctgatcac
     8641 tcagcggaag ttcgacaatc tcactaaggc tgagaggggc ggactgagcg aactggacaa
     8701 agcaggattc attaaacggc aacttgtgga gactcggcag attactaaac atgtcgccca
     8761 aatccttgac tcacgcatga ataccaagta cgacgaaaac gacaaactta tccgcgaggt
     8821 gaaggtgatt accctgaagt ccaagctggt cagcgatttc agaaaggact ttcaattcta
     8881 caaagtgcgg gagatcaata actatcatca tgctcatgac gcatatctga atgccgtggt
     8941 gggaaccgcc ctgatcaaga agtacccaaa gctggaaagc gagttcgtgt acggagacta
     9001 caaggtctac gacgtgcgca agatgattgc caaatctgag caggagatcg gaaaggccac
     9061 cgcaaagtac ttcttctaca gcaacatcat gaatttcttc aagaccgaaa tcacccttgc
     9121 aaacggtgag atccggaaga ggccgctcat cgagactaat ggggagactg gcgaaatcgt
     9181 gtgggacaag ggcagagatt tcgctaccgt gcgcaaagtg ctttctatgc ctcaagtgaa
     9241 catcgtgaag aaaaccgagg tgcaaaccgg aggcttttct aaggaatcaa tcctccccaa
     9301 gcgcaactcc gacaagctca ttgcaaggaa gaaggattgg gaccctaaga agtacggcgg
     9361 attcgattca ccaactgtgg cttattctgt cctggtcgtg gctaaggtgg aaaaaggaaa
     9421 gtctaagaag ctcaagagcg tgaaggaact gctgggtatc accattatgg agcgcagctc
     9481 cttcgagaag aacccaattg actttctcga agccaaaggt tacaaggaag tcaagaagga
     9541 ccttatcatc aagctcccaa agtatagcct gttcgaactg gagaatgggc ggaagcggat
     9601 gctcgcctcc gctggcgaac ttcagaaggg taatgagctg gctctcccct ccaagtacgt
     9661 gaatttcctc taccttgcaa gccattacga gaagctgaag gggagccccg aggacaacga
     9721 gcaaaagcaa ctgtttgtgg agcagcataa gcattatctg gacgagatca ttgagcagat
     9781 ttccgagttt tctaaacgcg tcattctcgc tgatgccaac ctcgataaag tccttagcgc
     9841 atacaataag cacagagaca aaccaattcg ggagcaggct gagaatatca tccacctgtt
     9901 caccctcacc aatcttggtg cccctgccgc attcaagtac ttcgacacca ccatcgaccg
     9961 gaaacgctat acctccacca aagaagtgct ggacgccacc ctcatccacc agagcatcac
    10021 cggactttac gaaactcgga ttgacctctc acagctcgga ggggatggtg gcggaggctc
    10081 gccaaaaaag aagagaaagg tagacccaaa gaaaaaacga aaagtagatc cgaaaaagaa
    10141 gaggaaggtg ggatccctgc ccacctgcag ctgtcttgat cgagttatac aaaaagacaa
    10201 aggcccatat tatacacacc ttggggcagg accaagtgtt gctgctgtca gggaaatcat
    10261 ggagaatagg tatggtcaaa aaggaaacgc aataaggata gaaatagtag tgtacaccgg
    10321 taaagaaggg aaaagctctc atgggtgtcc aattgctaag tgggttttaa gaagaagcag
    10381 tgatgaagaa aaagttcttt gtttggtccg gcagcgtaca ggccaccact gtccaactgc
    10441 tgtgatggtg gtgctcatca tggtgtggga tggcatccct cttccaatgg ccgaccggct
    10501 atacacagag ctcacagaga atctaaagtc atacaatggg caccctaccg acagaagatg
    10561 caccctcaat gaaaatcgta cctgtacatg tcaaggaatt gatccagaga cttgtggagc
    10621 ttcattctct tttggctgtt catggagtat gtactttaat ggctgtaagt ttggtagaag
    10681 cccaagcccc agaagattta gaattgatcc aagctctccc ttacatgaaa aaaaccttga
    10741 agataactta cagagtttgg ctacacgatt agctccaatt tataagcagt atgctccagt
    10801 agcttaccaa aatcaggtgg aatatgaaaa tgttgcccga gaatgtcggc ttggcagcaa
    10861 ggaaggtcga cccttctctg gggtcactgc ttgcctggac ttctgtgctc atccccacag
    10921 ggacattcac aacatgaata atggaagcac tgtggtttgt accttaactc gagaagataa
    10981 ccgctctttg ggtgttattc ctcaagatga gcagctccat gtgctacctc tttataagct
    11041 ttcagacaca gatgagtttg gctccaagga aggaatggaa gccaagatca aatctggggc
    11101 catcgaggtc ctggcacccc gccgcaaaaa aagaacgtgt ttcactcagc ctgttccccg
    11161 ttctggaaag aagagggctg cgatgatgac agaggttctt gcacataaga taagggcagt
    11221 ggaaaagaaa cctattcccc gaatcaagcg gaagaataac tcaacaacaa caaacaacag
    11281 taagccttcg tcactgccaa ccttagggag taacactgag accgtgcaac ctgaagtaaa
    11341 aagtgaaacc gaaccccatt ttatcttaaa aagttcagac aacactaaaa cttattcgct
    11401 gatgccatcc gctcctcacc cagtgaaaga ggcatctcca ggcttctcct ggtccccgaa
    11461 gactgcttca gccacaccag ctccactgaa gaatgacgca acagcctcat gcgggttttc
    11521 agaaagaagc agcactcccc actgtacgat gccttcggga agactcagtg gtgccaatgc
    11581 tgcagctgct gatggccctg gcatttcaca gcttggcgaa gtggctcctc tccccaccct
    11641 gtctgctcct gtgatggagc ccctcattaa ttctgagcct tccactggtg tgactgagcc
    11701 gctaacgcct catcagccaa accaccagcc ctccttcctc acctctcctc aagaccttgc
    11761 ctcttctcca atggaagaag atgagcagca ttctgaagca gatgagcctc catcagacga
    11821 acccctatct gatgaccccc tgtcacctgc tgaggagaaa ttgccccaca ttgatgagta
    11881 ttggtcagac agtgagcaca tctttttgga tgcaaatatt ggtggggtgg ccatcgcacc
    11941 tgctcacggc tcggttttga ttgagtgtgc ccggcgagag ctgcacgcta ccactcctgt
    12001 tgagcacccc aaccgtaatc atccaacccg cctctccctt gtcttttacc agcacaaaaa
    12061 cctaaataag ccccaacatg gttttgaact aaacaagatt aagtttgagg ctaaagaagc
    12121 taagaataag aaaatgaagg cctcagagca aaaagaccag gcagctaatg aaggtccaga
    12181 acagtcctct gaagtaaatg aattgaacca aattccttct cataaagcat taacattaac
    12241 ccatgacaat gttgtcaccg tgtcccctta tgctctcaca cacgttgcgg ggccctataa
    12301 ccattgggtc gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg
    12361 cccagtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
    12421 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
    12481 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
    12541 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
    12601 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
    12661 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
    12721 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
    12781 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
    12841 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
    12901 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
    12961 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
    13021 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagga
    13081 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
    13141 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
    13201 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
    13261 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
    13321 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
    13381 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
    13441 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
    13501 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
    13561 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
    13621 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
    13681 cgcctccccg catcgatacc gtcgacctcg agacctagaa aaacatggag caatcacaag
    13741 tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga
    13801 ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt
    13861 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
    13921 aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattggca
    13981 gaactacaca ccagggccag ggatcagata tccactgacc tttggatggt gctacaagct
    14041 agtaccagtt gagcaagaga aggtagaaga agccaatgaa ggagagaaca cccgcttgtt
    14101 acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtattag agtggaggtt
    14161 tgacagccgc ctagcatttc atcacatggc ccgagagctg catccggact gtactgggtc
    14221 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
    14281 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
    14341 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagggc
    14401 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
    14461 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
    14521 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
    14581 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
    14641 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
    14701 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
    14761 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
    14821 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
    14881 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
    14941 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
    15001 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
    15061 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
    15121 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
    15181 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
    15241 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
    15301 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
    15361 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
    15421 cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata
    15481 gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg
    15541 ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc
    15601 cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc
    15661 agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc
    15721 ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc
    15781 gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac
    15841 ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat
    15901 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
    15961 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
    16021 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
    16081 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
    16141 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
    16201 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
    16261 aaagcc
//
