LOCUS       sequence-462959-       16268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral cassette that expresses Tet1c-dCas9 fusion with a short 
            linker in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16268)
  AUTHORS   Rohm D, Black JB, McCutcheon SR, Barrera A, Berry SS, Morone DJ, 
            Nuttle X, de Esch CE, Tai DJC, Talkowski ME, Iglesias N, Gersbach CA
  TITLE     Activation of the imprinted Prader-Willi syndrome locus by 
            CRISPR-based epigenome editing.
  JOURNAL   Cell Genom. 2025 Feb 12;5(2):100770. doi: 
            10.1016/j.xgen.2025.100770.
  PUBMED    39947136
REFERENCE   2  (bases 1 to 16268)
  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: "Cell 
            Genom."; date: "2025-02-12"; volume: "5(2):100770. doi"; pages: " 
            10.1016/j.xgen.2025.100770"
COMMENT     Sequence Label: pLV-UbC-Tet1c-dCas9
FEATURES             Location/Qualifiers
     source          1..16268
                     /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"
     protein_bind    complement(2599..2632)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2656..3867
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3704..3723
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      3896..3905
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3908..3928
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6134..6199
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6206..6226
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(6251..6274,6275..6277,6278..8764,8765..8767,
                     8768..10351)
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6251..10351
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             10352..10399
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             10421..10474
                     /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             10481..10876
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     protein_bind    complement(10902..10935)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    10960..11548
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11013..11033)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11431..11442)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(11551..11567)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11552..11568)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             12073..12253
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(12279..12296)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12285..12509
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      12555..12983
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12642..12661)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12852..12873
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12992..13012)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12997..13326
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      13177..13312
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     13239..13258
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        13374..13421
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             13440..13814
                     /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    13944..14065
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13981..14000)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     14035..14054
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(14114..14130)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14114..14130)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14127..14149)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14138..14154
                     /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(14162..14168,14169..14186,14187..14192))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14207..14228
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(14345..14362)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14516..15104)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14596..14615)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(15275..16066,16067..16135))
                     /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     15898..15917
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(16136..16240)
                     /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 tggttaatat aacttcgtat agcatacatt atacgaagtt attaacccgt
     2641 gtcggctcca gatctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcc
     2701 tcacggcgag cgctgccacg tcagacgaag ggcgcagcga gcgtcctgat ccttccgccc
     2761 ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc
     2821 agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga
     2881 gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg
     2941 gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc
     3001 agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta
     3061 gcgggctgct gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc
     3121 gtgtggagag accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg
     3181 gggttggggg gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg
     3241 tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca
     3301 aggtcttgag gccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctggggca
     3361 ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt tgtcgtctgt
     3421 tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg
     3481 cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac
     3541 ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg
     3601 taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt
     3661 cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga
     3721 actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg
     3781 taatcatttg ggtcaatatg taattttcag tgttagacta gtaaattgtc cgctaaattc
     3841 tggccgtttt tggctttttt gttagacgaa gcttgggctg caggtcgact ctagagccac
     3901 catggtgcca aaaaagaaga gaaaggtagg cggaggcggg agcggatccc tgcccacctg
     3961 cagctgtctt gatcgagtta tacaaaaaga caaaggccca tattatacac accttggggc
     4021 aggaccaagt gttgctgctg tcagggaaat catggagaat aggtatggtc aaaaaggaaa
     4081 cgcaataagg atagaaatag tagtgtacac cggtaaagaa gggaaaagct ctcatgggtg
     4141 tccaattgct aagtgggttt taagaagaag cagtgatgaa gaaaaagttc tttgtttggt
     4201 ccggcagcgt acaggccacc actgtccaac tgctgtgatg gtggtgctca tcatggtgtg
     4261 ggatggcatc cctcttccaa tggccgaccg gctatacaca gagctcacag agaatctaaa
     4321 gtcatacaat gggcacccta ccgacagaag atgcaccctc aatgaaaatc gtacctgtac
     4381 atgtcaagga attgatccag agacttgtgg agcttcattc tcttttggct gttcatggag
     4441 tatgtacttt aatggctgta agtttggtag aagcccaagc cccagaagat ttagaattga
     4501 tccaagctct cccttacatg aaaaaaacct tgaagataac ttacagagtt tggctacacg
     4561 attagctcca atttataagc agtatgctcc agtagcttac caaaatcagg tggaatatga
     4621 aaatgttgcc cgagaatgtc ggcttggcag caaggaaggt cgacccttct ctggggtcac
     4681 tgcttgcctg gacttctgtg ctcatcccca cagggacatt cacaacatga ataatggaag
     4741 cactgtggtt tgtaccttaa ctcgagaaga taaccgctct ttgggtgtta ttcctcaaga
     4801 tgagcagctc catgtgctac ctctttataa gctttcagac acagatgagt ttggctccaa
     4861 ggaaggaatg gaagccaaga tcaaatctgg ggccatcgag gtcctggcac cccgccgcaa
     4921 aaaaagaacg tgtttcactc agcctgttcc ccgttctgga aagaagaggg ctgcgatgat
     4981 gacagaggtt cttgcacata agataagggc agtggaaaag aaacctattc cccgaatcaa
     5041 gcggaagaat aactcaacaa caacaaacaa cagtaagcct tcgtcactgc caaccttagg
     5101 gagtaacact gagaccgtgc aacctgaagt aaaaagtgaa accgaacccc attttatctt
     5161 aaaaagttca gacaacacta aaacttattc gctgatgcca tccgctcctc acccagtgaa
     5221 agaggcatct ccaggcttct cctggtcccc gaagactgct tcagccacac cagctccact
     5281 gaagaatgac gcaacagcct catgcgggtt ttcagaaaga agcagcactc cccactgtac
     5341 gatgccttcg ggaagactca gtggtgccaa tgctgcagct gctgatggcc ctggcatttc
     5401 acagcttggc gaagtggctc ctctccccac cctgtctgct cctgtgatgg agcccctcat
     5461 taattctgag ccttccactg gtgtgactga gccgctaacg cctcatcagc caaaccacca
     5521 gccctccttc ctcacctctc ctcaagacct tgcctcttct ccaatggaag aagatgagca
     5581 gcattctgaa gcagatgagc ctccatcaga cgaaccccta tctgatgacc ccctgtcacc
     5641 tgctgaggag aaattgcccc acattgatga gtattggtca gacagtgagc acatcttttt
     5701 ggatgcaaat attggtgggg tggccatcgc acctgctcac ggctcggttt tgattgagtg
     5761 tgcccggcga gagctgcacg ctaccactcc tgttgagcac cccaaccgta atcatccaac
     5821 ccgcctctcc cttgtctttt accagcacaa aaacctaaat aagccccaac atggttttga
     5881 actaaacaag attaagtttg aggctaaaga agctaagaat aagaaaatga aggcctcaga
     5941 gcaaaaagac caggcagcta atgaaggtcc agaacagtcc tctgaagtaa atgaattgaa
     6001 ccaaattcct tctcataaag cattaacatt aacccatgac aatgttgtca ccgtgtcccc
     6061 ttatgctctc acacacgttg cggggcccta taaccattgg gtcggccggc cattcggtgg
     6121 aggtggctcc atggactata aggaccacga cggagactac aaggatcatg atattgatta
     6181 caaagacgat gacgataaga tggccccaaa gaagaagcgg aaggtcggta tccacggagt
     6241 cccagcagcc gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg
     6301 ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac
     6361 cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac
     6421 agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg
     6481 gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt
     6541 ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat
     6601 cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct
     6661 gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct
     6721 ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa
     6781 cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga
     6841 aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa
     6901 gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt
     6961 cggaaacctg attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct
     7021 ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct
     7081 gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga
     7141 cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag
     7201 cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct
     7261 cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg
     7321 ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc
     7381 catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct
     7441 gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga
     7501 gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga
     7561 aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg
     7621 aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt
     7681 cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt
     7741 cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt
     7801 caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc
     7861 cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa
     7921 agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt
     7981 ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct
     8041 gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga
     8101 tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac
     8161 ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg
     8221 ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac
     8281 aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca
     8341 cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga
     8401 tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct
     8461 gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa
     8521 catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg
     8581 cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga
     8641 acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
     8701 tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt
     8761 ggacgccatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac
     8821 cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa
     8881 gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga
     8941 caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa
     9001 gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg
     9061 gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct
     9121 gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat
     9181 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat
     9241 caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt
     9301 gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt
     9361 ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg
     9421 gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg
     9481 ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac
     9541 cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa
     9601 gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac
     9661 cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa
     9721 gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc
     9781 catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct
     9841 gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg
     9901 cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct
     9961 ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt
    10021 tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa
    10081 gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg
    10141 ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
    10201 gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag
    10261 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
    10321 acggatcgac ctgtctcagc tgggaggcga caaaaggccg gcggccacga aaaaggccgg
    10381 acaggccaaa aagaaaaagg gcggagattt cggcgctagc gagggcagag gaagtcttct
    10441 aacatgcggt gacgtggagg agaatcccgg ccctggtacc atggccaagc ctttgtctca
    10501 agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga
    10561 agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa
    10621 tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc
    10681 tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt
    10741 gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat
    10801 agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg
    10861 ttatgtgtgg gagggcctgc agctgcagta gttgataata gataacttcg tatagcatac
    10921 attatacgaa gttatgaatt cgatatcaag cttatcgata atcaacctct ggattacaaa
    10981 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
    11041 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
    11101 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
    11161 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
    11221 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
    11281 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
    11341 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
    11401 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
    11461 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
    11521 cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgaga cctagaaaaa
    11581 catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa
    11641 gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg
    11701 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
    11761 ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc
    11821 tacttccctg attggcagaa ctacacacca gggccaggga tcagatatcc actgaccttt
    11881 ggatggtgct acaagctagt accagttgag caagagaagg tagaagaagc caatgaagga
    11941 gagaacaccc gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa
    12001 gtattagagt ggaggtttga cagccgccta gcatttcatc acatggcccg agagctgcat
    12061 ccggactgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
    12121 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
    12181 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
    12241 aaaatctcta gcagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
    12301 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
    12361 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
    12421 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
    12481 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
    12541 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
    12601 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
    12661 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
    12721 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
    12781 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
    12841 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
    12901 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
    12961 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
    13021 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
    13081 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
    13141 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
    13201 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
    13261 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
    13321 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca
    13381 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca
    13441 tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg
    13501 agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac ttcgccggtg
    13561 tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca
    13621 acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag tggtcggagg
    13681 tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc ggcgagcagc
    13741 cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg
    13801 aggagcagga ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt
    13861 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
    13921 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
    13981 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
    14041 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
    14101 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
    14161 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
    14221 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
    14281 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
    14341 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
    14401 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
    14461 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
    14521 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
    14581 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
    14641 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
    14701 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
    14761 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
    14821 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
    14881 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
    14941 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
    15001 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
    15061 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
    15121 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
    15181 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
    15241 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
    15301 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
    15361 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
    15421 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
    15481 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
    15541 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
    15601 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
    15661 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
    15721 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
    15781 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
    15841 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
    15901 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
    15961 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
    16021 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
    16081 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
    16141 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
    16201 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
    16261 cacctgac
//
