LOCUS       sequence-381312-       16256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral construct to express spdCas9-Tet1-GFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16256)
  AUTHORS   Pahlevan Kakhki M, Giordano A, Starvaggi Cucuzza C, Venkata S Badam 
            T, Samudyata S, Lemee MV, Stridh P, Gkogka A, Shchetynsky K, Harroud
            A, Gyllenberg A, Liu Y, Boddul S, James T, Sorosina M, Filippi M, 
            Esposito F, Wermeling F, Gustafsson M, Casaccia P, Hillert J, Olsson
            T, Kockum I, Sellgren CM, Golzio C, Kular L, Jagodic M
  TITLE     A genetic-epigenetic interplay at 1q21.1 locus underlies 
            CHD1L-mediated vulnerability to primary progressive multiple 
            sclerosis.
  JOURNAL   Nat Commun. 2024 Jul 30;15(1):6419. doi: 10.1038/s41467-024-50794-z.
  PUBMED    39079955
REFERENCE   2  (bases 1 to 16256)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2024-07-30"; volume: "15(1):6419. doi"; pages: " 
            10.1038/s41467-024-50794-z"
COMMENT     Sequence Label: P3-Lenti-dCas9-Tet1-GFP
FEATURES             Location/Qualifiers
     source          1..16256
                     /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(join(1080..1871,1872..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."
     CDS             4171..4215
                     /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    4485..4602
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4667..5878
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5715..5734
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(5935..5961,5962..5964,5965..8451,8452..8454,
                     8455..10038)
                     /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             10054..10074
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10078..10098
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10102..10122
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             12283..12336
                     /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             join(12337..12339,12340..13050)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(12337..12339,12340..13050)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(12379..12400)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(12640..12659)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     12987..13008
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    13093..13681
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(13146..13166)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(13564..13575)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(13684..13700)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13685..13701)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             14206..14386
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(14412..14429)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    14418..14642
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      14688..15116
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14775..14794)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14985..15006
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(15125..15145)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        15169..15379
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        15191..15387
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      15238..15373
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     15300..15319
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        15435..15482
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             15501..15875
                     /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    16005..16126
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(16042..16061)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     16096..16115
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(16175..16191)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(16175..16191)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(16188..16210)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    16199..16215
                     /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(16223..16229,16230..16247,16248..16253))
                     /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 atcccaaccc cgaggggacc cgacaggccc gaaggaatag
     4381 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac
     4441 tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg
     4501 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     4561 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
     4621 agcagagatc cagtttggtt aattaacccg tgtcggctcc agatctggcc tccgcgccgg
     4681 gttttggcgc ctcccgcggg cgcccccctc ctcacggcga gcgctgccac gtcagacgaa
     4741 gggcgcagcg agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca
     4801 taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt
     4861 gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt
     4921 ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac
     4981 gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt
     5041 gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg
     5101 tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc
     5161 tgggtccgcg agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg
     5221 ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa
     5281 ggtggggggc atggtgggcg gcaagaaccc aaggtcttga ggccttcgct aatgcgggaa
     5341 agctcttatt cgggtgagat gggctggggc accatctggg gaccctgacg tgaagtttgt
     5401 cactgactgg agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt
     5461 tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt
     5521 tctgttggct tataatgcag ggtggggcca cctgccggta ggtgtgcggt aggcttttct
     5581 ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga
     5641 cctctggtga ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta
     5701 tcttcttaag tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt
     5761 ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca
     5821 gtgttagact agtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacga
     5881 agcttgggct gcaggtcgac tctagaggat ccagttaacc tcgagggcgc gcccatggac
     5941 aagaagtatt ctatcggact ggccatcggg actaatagcg tcgggtgggc cgtgatcact
     6001 gacgagtaca aggtgccctc taagaagttc aaggtgctcg ggaacaccga ccggcattcc
     6061 atcaagaaaa atctgatcgg agctctcctc tttgattcag gggagaccgc tgaagcaacc
     6121 cgcctcaagc ggactgctag acggcggtac accaggagga agaaccggat ttgttacctt
     6181 caagagatat tctccaacga aatggcaaag gtcgacgaca gcttcttcca taggctggaa
     6241 gaatcattcc tcgtggaaga ggataagaag catgaacggc atcccatctt cggtaatatc
     6301 gtcgacgagg tggcctatca cgagaaatac ccaaccatct accatcttcg caaaaagctg
     6361 gtggactcaa ccgacaaggc agacctccgg cttatctacc tggccctggc ccacatgatc
     6421 aagttcagag gccacttcct gatcgagggc gacctcaatc ctgacaatag cgatgtggat
     6481 aaactgttca tccagctggt gcagacttac aaccagctct ttgaagagaa ccccatcaat
     6541 gcaagcggag tcgatgccaa ggccattctg tcagcccggc tgtcaaagag ccgcagactt
     6601 gagaatctta tcgctcagct gccgggtgaa aagaaaaatg gactgttcgg gaacctgatt
     6661 gctctttcac ttgggctgac tcccaatttc aagtctaatt tcgacctggc agaggatgcc
     6721 aagctgcaac tgtccaagga cacctatgat gacgatctcg acaacctcct ggcccagatc
     6781 ggtgaccaat acgccgacct tttccttgct gctaagaatc tttctgacgc catcctgctg
     6841 tctgacattc tccgcgtgaa cactgaaatc accaaggccc ctctttcagc ttcaatgatt
     6901 aagcggtatg atgagcacca ccaggacctg accctgctta aggcactcgt ccggcagcag
     6961 cttccggaga agtacaagga aatcttcttt gaccagtcaa agaatggata cgccggctac
     7021 atcgacggag gtgcctccca agaggaattt tataagttta tcaaacctat ccttgagaag
     7081 atggacggca ccgaagagct cctcgtgaaa ctgaatcggg aggatctgct gcggaagcag
     7141 cgcactttcg acaatgggag cattccccac cagatccatc ttggggagct tcacgccatc
     7201 cttcggcgcc aagaggactt ctaccccttt cttaaggaca acagggagaa gattgagaaa
     7261 attctcactt tccgcatccc ctactacgtg ggacccctcg ccagaggaaa tagccggttt
     7321 gcttggatga ccagaaagtc agaagaaact atcactccct ggaacttcga agaggtggtg
     7381 gacaagggag ccagcgctca gtcattcatc gaacggatga ctaacttcga taagaacctc
     7441 cccaatgaga aggtcctgcc gaaacattcc ctgctctacg agtactttac cgtgtacaac
     7501 gagctgacca aggtgaaata tgtcaccgaa gggatgagga agcccgcatt cctgtcaggc
     7561 gaacaaaaga aggcaattgt ggaccttctg ttcaagacca atagaaaggt gaccgtgaag
     7621 cagctgaagg aggactattt caagaaaatt gaatgcttcg actctgtgga gattagcggg
     7681 gtcgaagatc ggttcaacgc aagcctgggt acctaccatg atctgcttaa gatcatcaag
     7741 gacaaggatt ttctggacaa tgaggagaac gaggacatcc ttgaggacat tgtcctgact
     7801 ctcactctgt tcgaggaccg ggaaatgatc gaggagaggc ttaagaccta cgcccatctg
     7861 ttcgacgata aagtgatgaa gcaacttaaa cggagaagat ataccggatg gggacgcctt
     7921 agccgcaaac tcatcaacgg aatccgggac aaacagagcg gaaagaccat tcttgatttc
     7981 cttaaaagcg acggattcgc taatcgcaac ttcatgcaac ttatccatga tgattccctg
     8041 acctttaagg aggacatcca gaaggcccaa gtgtctggac aaggtgactc actgcacgag
     8101 catatcgcaa atctggctgg ttcacccgct attaagaagg gtattctcca gaccgtgaaa
     8161 gtcgtggacg agctggtcaa ggtgatgggt cgccataaac cagagaacat tgtcatcgag
     8221 atggccaggg aaaaccagac tacccagaag ggacagaaga acagcaggga gcggatgaaa
     8281 agaattgagg aagggattaa ggagctcggg tcacagatcc ttaaagagca cccggtggaa
     8341 aacacccagc ttcagaatga gaagctctat ctgtactacc ttcaaaatgg acgcgatatg
     8401 tatgtggacc aagagcttga tatcaacagg ctctcagact acgacgtgga cgccatcgtc
     8461 cctcagagct tcctcaaaga cgactcaatt gacaataagg tgctgactcg ctcagacaag
     8521 aaccggggaa agtcagataa cgtgccctca gaggaagtcg tgaaaaagat gaagaactat
     8581 tggcgccagc ttctgaacgc aaagctgatc actcagcgga agttcgacaa tctcactaag
     8641 gctgagaggg gcggactgag cgaactggac aaagcaggat tcattaaacg gcaacttgtg
     8701 gagactcggc agattactaa acatgtcgcc caaatccttg actcacgcat gaataccaag
     8761 tacgacgaaa acgacaaact tatccgcgag gtgaaggtga ttaccctgaa gtccaagctg
     8821 gtcagcgatt tcagaaagga ctttcaattc tacaaagtgc gggagatcaa taactatcat
     8881 catgctcatg acgcatatct gaatgccgtg gtgggaaccg ccctgatcaa gaagtaccca
     8941 aagctggaaa gcgagttcgt gtacggagac tacaaggtct acgacgtgcg caagatgatt
     9001 gccaaatctg agcaggagat cggaaaggcc accgcaaagt acttcttcta cagcaacatc
     9061 atgaatttct tcaagaccga aatcaccctt gcaaacggtg agatccggaa gaggccgctc
     9121 atcgagacta atggggagac tggcgaaatc gtgtgggaca agggcagaga tttcgctacc
     9181 gtgcgcaaag tgctttctat gcctcaagtg aacatcgtga agaaaaccga ggtgcaaacc
     9241 ggaggctttt ctaaggaatc aatcctcccc aagcgcaact ccgacaagct cattgcaagg
     9301 aagaaggatt gggaccctaa gaagtacggc ggattcgatt caccaactgt ggcttattct
     9361 gtcctggtcg tggctaaggt ggaaaaagga aagtctaaga agctcaagag cgtgaaggaa
     9421 ctgctgggta tcaccattat ggagcgcagc tccttcgaga agaacccaat tgactttctc
     9481 gaagccaaag gttacaagga agtcaagaag gaccttatca tcaagctccc aaagtatagc
     9541 ctgttcgaac tggagaatgg gcggaagcgg atgctcgcct ccgctggcga acttcagaag
     9601 ggtaatgagc tggctctccc ctccaagtac gtgaatttcc tctaccttgc aagccattac
     9661 gagaagctga aggggagccc cgaggacaac gagcaaaagc aactgtttgt ggagcagcat
     9721 aagcattatc tggacgagat cattgagcag atttccgagt tttctaaacg cgtcattctc
     9781 gctgatgcca acctcgataa agtccttagc gcatacaata agcacagaga caaaccaatt
     9841 cgggagcagg ctgagaatat catccacctg ttcaccctca ccaatcttgg tgcccctgcc
     9901 gcattcaagt acttcgacac caccatcgac cggaaacgct atacctccac caaagaagtg
     9961 ctggacgcca ccctcatcca ccagagcatc accggacttt acgaaactcg gattgacctc
    10021 tcacagctcg gaggggatgg tggcggaggc tcgccaaaaa agaagagaaa ggtagaccca
    10081 aagaaaaaac gaaaagtaga tccgaaaaag aagaggaagg tgggatccct gcccacctgc
    10141 agctgtcttg atcgagttat acaaaaagac aaaggcccat attatacaca ccttggggca
    10201 ggaccaagtg ttgctgctgt cagggaaatc atggagaata ggtatggtca aaaaggaaac
    10261 gcaataagga tagaaatagt agtgtacacc ggtaaagaag ggaaaagctc tcatgggtgt
    10321 ccaattgcta agtgggtttt aagaagaagc agtgatgaag aaaaagttct ttgtttggtc
    10381 cggcagcgta caggccacca ctgtccaact gctgtgatgg tggtgctcat catggtgtgg
    10441 gatggcatcc ctcttccaat ggccgaccgg ctatacacag agctcacaga gaatctaaag
    10501 tcatacaatg ggcaccctac cgacagaaga tgcaccctca atgaaaatcg tacctgtaca
    10561 tgtcaaggaa ttgatccaga gacttgtgga gcttcattct cttttggctg ttcatggagt
    10621 atgtacttta atggctgtaa gtttggtaga agcccaagcc ccagaagatt tagaattgat
    10681 ccaagctctc ccttacatga aaaaaacctt gaagataact tacagagttt ggctacacga
    10741 ttagctccaa tttataagca gtatgctcca gtagcttacc aaaatcaggt ggaatatgaa
    10801 aatgttgccc gagaatgtcg gcttggcagc aaggaaggtc gacccttctc tggggtcact
    10861 gcttgcctgg acttctgtgc tcatccccac agggacattc acaacatgaa taatggaagc
    10921 actgtggttt gtaccttaac tcgagaagat aaccgctctt tgggtgttat tcctcaagat
    10981 gagcagctcc atgtgctacc tctttataag ctttcagaca cagatgagtt tggctccaag
    11041 gaaggaatgg aagccaagat caaatctggg gccatcgagg tcctggcacc ccgccgcaaa
    11101 aaaagaacgt gtttcactca gcctgttccc cgttctggaa agaagagggc tgcgatgatg
    11161 acagaggttc ttgcacataa gataagggca gtggaaaaga aacctattcc ccgaatcaag
    11221 cggaagaata actcaacaac aacaaacaac agtaagcctt cgtcactgcc aaccttaggg
    11281 agtaacactg agaccgtgca acctgaagta aaaagtgaaa ccgaacccca ttttatctta
    11341 aaaagttcag acaacactaa aacttattcg ctgatgccat ccgctcctca cccagtgaaa
    11401 gaggcatctc caggcttctc ctggtccccg aagactgctt cagccacacc agctccactg
    11461 aagaatgacg caacagcctc atgcgggttt tcagaaagaa gcagcactcc ccactgtacg
    11521 atgccttcgg gaagactcag tggtgccaat gctgcagctg ctgatggccc tggcatttca
    11581 cagcttggcg aagtggctcc tctccccacc ctgtctgctc ctgtgatgga gcccctcatt
    11641 aattctgagc cttccactgg tgtgactgag ccgctaacgc ctcatcagcc aaaccaccag
    11701 ccctccttcc tcacctctcc tcaagacctt gcctcttctc caatggaaga agatgagcag
    11761 cattctgaag cagatgagcc tccatcagac gaacccctat ctgatgaccc cctgtcacct
    11821 gctgaggaga aattgcccca cattgatgag tattggtcag acagtgagca catctttttg
    11881 gatgcaaata ttggtggggt ggccatcgca cctgctcacg gctcggtttt gattgagtgt
    11941 gcccggcgag agctgcacgc taccactcct gttgagcacc ccaaccgtaa tcatccaacc
    12001 cgcctctccc ttgtctttta ccagcacaaa aacctaaata agccccaaca tggttttgaa
    12061 ctaaacaaga ttaagtttga ggctaaagaa gctaagaata agaaaatgaa ggcctcagag
    12121 caaaaagacc aggcagctaa tgaaggtcca gaacagtcct ctgaagtaaa tgaattgaac
    12181 caaattcctt ctcataaagc attaacatta acccatgaca atgttgtcac cgtgtcccct
    12241 tatgctctca cacacgttgc ggggccctat aaccattggg tcgagggcag aggaagtctg
    12301 ctaacatgcg gtgacgtcga ggagaatcct ggcccagtga gcaagggcga ggagctgttc
    12361 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
    12421 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
    12481 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
    12541 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
    12601 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
    12661 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
    12721 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
    12781 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
    12841 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
    12901 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
    12961 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
    13021 atcactctcg gcatggacga gctgtacaag gaattcaatt ctaactagga attcgatatc
    13081 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
    13141 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
    13201 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
    13261 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
    13321 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
    13381 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
    13441 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
    13501 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
    13561 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
    13621 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
    13681 catcgatacc gtcgacctcg agacctagaa aaacatggag caatcacaag tagcaataca
    13741 gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt
    13801 ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
    13861 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
    13921 atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattggca gaactacaca
    13981 ccagggccag ggatcagata tccactgacc tttggatggt gctacaagct agtaccagtt
    14041 gagcaagaga aggtagaaga agccaatgaa ggagagaaca cccgcttgtt acaccctgtg
    14101 agcctgcatg ggatggatga cccggagaga gaagtattag agtggaggtt tgacagccgc
    14161 ctagcatttc atcacatggc ccgagagctg catccggact gtactgggtc tctctggtta
    14221 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
    14281 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
    14341 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagggc ccgtttaaac
    14401 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
    14461 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
    14521 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
    14581 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
    14641 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
    14701 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
    14761 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
    14821 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
    14881 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
    14941 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
    15001 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
    15061 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
    15121 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
    15181 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
    15241 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
    15301 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
    15361 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
    15421 tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg
    15481 acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc
    15541 gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg
    15601 tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc
    15661 aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc
    15721 tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca
    15781 tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca
    15841 actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca
    15901 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
    15961 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
    16021 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
    16081 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
    16141 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
    16201 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcc
//
