LOCUS       sequence-462961-       16309 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet1v4-dCas9 from Nunez et al. 2021, in a lentiviral cassette with 
            blasticidin resistance, for expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16309)
  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 16309)
  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-Tet1v4-dCas9
FEATURES             Location/Qualifiers
     source          1..16309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2622..3833
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3670..3689
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      3862..3871
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6268..6294,6295..6297,6298..8784,8785..8787,
                     8788..10371)
                     /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             10387..10407
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10411..10431
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10435..10455
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10462..10515
                     /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             10522..10917
                     /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(10943..10976)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    11001..11589
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11054..11074)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11472..11483)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(11592..11608)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11593..11609)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             12114..12294
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(12320..12337)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12326..12550
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      12596..13024
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12683..12702)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12893..12914
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(13033..13053)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        13038..13367
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      13218..13353
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     13280..13299
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        13415..13462
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             13481..13855
                     /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    13985..14106
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(14022..14041)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     14076..14095
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(14155..14171)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14155..14171)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14168..14190)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14179..14195
                     /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(14203..14209,14210..14227,14228..14233))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14248..14269
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(14386..14403)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14557..15145)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14637..14656)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(15316..16107,16108..16176))
                     /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     15939..15958
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(16177..16281)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta acccgtgtcg gctccagatc tggcctccgc gccgggtttt
     2641 ggcgcctccc gcgggcgccc ccctcctcac ggcgagcgct gccacgtcag acgaagggcg
     2701 cagcgagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga
     2761 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc
     2821 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg
     2881 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc
     2941 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga
     3001 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc
     3061 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt
     3121 ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt
     3181 cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg
     3241 ggggcatggt gggcggcaag aacccaaggt cttgaggcct tcgctaatgc gggaaagctc
     3301 ttattcgggt gagatgggct ggggcaccat ctggggaccc tgacgtgaag tttgtcactg
     3361 actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt gccgttgggc
     3421 agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt
     3481 tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct tttctccgtc
     3541 gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg ccggacctct
     3601 ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt acctatcttc
     3661 ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg
     3721 aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat tttcagtgtt
     3781 agactagtaa attgtccgct aaattctggc cgtttttggc ttttttgtta gacgaagctt
     3841 gggctgcagg tcgactctag agccaccatg gccctgccca cctgcagctg tcttgatcga
     3901 gttatacaaa aagacaaagg cccatattat acacaccttg gggcaggacc aagtgttgct
     3961 gctgtcaggg aaatcatgga gaataggtat ggtcaaaaag gaaacgcaat aaggatagaa
     4021 atagtagtgt acaccggtaa agaagggaaa agctctcatg ggtgtccaat tgctaagtgg
     4081 gttttaagaa gaagcagtga tgaagaaaaa gttctttgtt tggtccggca gcgtacaggc
     4141 caccactgtc caactgctgt gatggtggtg ctcatcatgg tgtgggatgg catccctctt
     4201 ccaatggccg accggctata cacagagctc acagagaatc taaagtcata caatgggcac
     4261 cctaccgaca gaagatgcac cctcaatgaa aatcgtacct gtacatgtca aggaattgat
     4321 ccagagactt gtggagcttc attctctttt ggctgttcat ggagtatgta ctttaatggc
     4381 tgtaagtttg gtagaagccc aagccccaga agatttagaa ttgatccaag ctctccctta
     4441 catgaaaaaa accttgaaga taacttacag agtttggcta cacgattagc tccaatttat
     4501 aagcagtatg ctccagtagc ttaccaaaat caggtggaat atgaaaatgt tgcccgagaa
     4561 tgtcggcttg gcagcaagga aggtcgaccc ttctctgggg tcactgcttg cctggacttc
     4621 tgtgctcatc cccacaggga cattcacaac atgaataatg gaagcactgt ggtttgtacc
     4681 ttaactcgag aagataaccg ctctttgggt gttattcctc aagatgagca gctccatgtg
     4741 ctacctcttt ataagctttc agacacagat gagtttggct ccaaggaagg aatggaagcc
     4801 aagatcaaat ctggggccat cgaggtcctg gcaccccgcc gcaaaaaaag aacgtgtttc
     4861 actcagcctg ttccccgttc tggaaagaag agggctgcga tgatgacaga ggttcttgca
     4921 cataagataa gggcagtgga aaagaaacct attccccgaa tcaagcggaa gaataactca
     4981 acaacaacaa acaacagtaa gccttcgtca ctgccaacct tagggagtaa cactgagacc
     5041 gtgcaacctg aagtaaaaag tgaaaccgaa ccccatttta tcttaaaaag ttcagacaac
     5101 actaaaactt attcgctgat gccatccgct cctcacccag tgaaagaggc atctccaggc
     5161 ttctcctggt ccccgaagac tgcttcagcc acaccagctc cactgaagaa tgacgcaaca
     5221 gcctcatgcg ggttttcaga aagaagcagc actccccact gtacgatgcc ttcgggaaga
     5281 ctcagtggtg ccaatgctgc agctgctgat ggccctggca tttcacagct tggcgaagtg
     5341 gctcctctcc ccaccctgtc tgctcctgtg atggagcccc tcattaattc tgagccttcc
     5401 actggtgtga ctgagccgct aacgcctcat cagccaaacc accagccctc cttcctcacc
     5461 tctcctcaag accttgcctc ttctccaatg gaagaagatg agcagcattc tgaagcagat
     5521 gagcctccat cagacgaacc cctatctgat gaccccctgt cacctgctga ggagaaattg
     5581 ccccacattg atgagtattg gtcagacagt gagcacatct ttttggatgc aaatattggt
     5641 ggggtggcca tcgcacctgc tcacggctcg gttttgattg agtgtgcccg gcgagagctg
     5701 cacgctacca ctcctgttga gcaccccaac cgtaatcatc caacccgcct ctcccttgtc
     5761 ttttaccagc acaaaaacct aaataagccc caacatggtt ttgaactaaa caagattaag
     5821 tttgaggcta aagaagctaa gaataagaaa atgaaggcct cagagcaaaa agaccaggca
     5881 gctaatgaag gtccagaaca gtcctctgaa gtaaatgaat tgaaccaaat tccttctcat
     5941 aaagcattaa cattaaccca tgacaatgtt gtcaccgtgt ccccttatgc tctcacacac
     6001 gttgcggggc cctataacca ttgggtcgga gggccgagct ctggcgcacc cccaccaagt
     6061 ggagggtctc ctgccgggtc cccaacatct actgaagaag gcaccagcga atccgcaacg
     6121 cccgagtcag gccctggtac ctccacagaa ccatctgaag gtagtgcgcc tggttcccca
     6181 gctggaagcc ctacttccac cgaagaaggc acgtcaaccg aaccaagtga aggatctgcc
     6241 cctgggacca gcactgaacc atctgagatg gacaagaagt attctatcgg actggccatc
     6301 gggactaata gcgtcgggtg ggccgtgatc actgacgagt acaaggtgcc ctctaagaag
     6361 ttcaaggtgc tcgggaacac cgaccggcat tccatcaaga aaaatctgat cggagctctc
     6421 ctctttgatt caggggagac cgctgaagca acccgcctca agcggactgc tagacggcgg
     6481 tacaccagga ggaagaaccg gatttgttac cttcaagaga tattctccaa cgaaatggca
     6541 aaggtcgacg acagcttctt ccataggctg gaagaatcat tcctcgtgga agaggataag
     6601 aagcatgaac ggcatcccat cttcggtaat atcgtcgacg aggtggccta tcacgagaaa
     6661 tacccaacca tctaccatct tcgcaaaaag ctggtggact caaccgacaa ggcagacctc
     6721 cggcttatct acctggccct ggcccacatg atcaagttca gaggccactt cctgatcgag
     6781 ggcgacctca atcctgacaa tagcgatgtg gataaactgt tcatccagct ggtgcagact
     6841 tacaaccagc tctttgaaga gaaccccatc aatgcaagcg gagtcgatgc caaggccatt
     6901 ctgtcagccc ggctgtcaaa gagccgcaga cttgagaatc ttatcgctca gctgccgggt
     6961 gaaaagaaaa atggactgtt cgggaacctg attgctcttt cacttgggct gactcccaat
     7021 ttcaagtcta atttcgacct ggcagaggat gccaagctgc aactgtccaa ggacacctat
     7081 gatgacgatc tcgacaacct cctggcccag atcggtgacc aatacgccga ccttttcctt
     7141 gctgctaaga atctttctga cgccatcctg ctgtctgaca ttctccgcgt gaacactgaa
     7201 atcaccaagg cccctctttc agcttcaatg attaagcggt atgatgagca ccaccaggac
     7261 ctgaccctgc ttaaggcact cgtccggcag cagcttccgg agaagtacaa ggaaatcttc
     7321 tttgaccagt caaagaatgg atacgccggc tacatcgacg gaggtgcctc ccaagaggaa
     7381 ttttataagt ttatcaaacc tatccttgag aagatggacg gcaccgaaga gctcctcgtg
     7441 aaactgaatc gggaggatct gctgcggaag cagcgcactt tcgacaatgg gagcattccc
     7501 caccagatcc atcttgggga gcttcacgcc atccttcggc gccaagagga cttctacccc
     7561 tttcttaagg acaacaggga gaagattgag aaaattctca ctttccgcat cccctactac
     7621 gtgggacccc tcgccagagg aaatagccgg tttgcttgga tgaccagaaa gtcagaagaa
     7681 actatcactc cctggaactt cgaagaggtg gtggacaagg gagccagcgc tcagtcattc
     7741 atcgaacgga tgactaactt cgataagaac ctccccaatg agaaggtcct gccgaaacat
     7801 tccctgctct acgagtactt taccgtgtac aacgagctga ccaaggtgaa atatgtcacc
     7861 gaagggatga ggaagcccgc attcctgtca ggcgaacaaa agaaggcaat tgtggacctt
     7921 ctgttcaaga ccaatagaaa ggtgaccgtg aagcagctga aggaggacta tttcaagaaa
     7981 attgaatgct tcgactctgt ggagattagc ggggtcgaag atcggttcaa cgcaagcctg
     8041 ggtacctacc atgatctgct taagatcatc aaggacaagg attttctgga caatgaggag
     8101 aacgaggaca tccttgagga cattgtcctg actctcactc tgttcgagga ccgggaaatg
     8161 atcgaggaga ggcttaagac ctacgcccat ctgttcgacg ataaagtgat gaagcaactt
     8221 aaacggagaa gatataccgg atggggacgc cttagccgca aactcatcaa cggaatccgg
     8281 gacaaacaga gcggaaagac cattcttgat ttccttaaaa gcgacggatt cgctaatcgc
     8341 aacttcatgc aacttatcca tgatgattcc ctgaccttta aggaggacat ccagaaggcc
     8401 caagtgtctg gacaaggtga ctcactgcac gagcatatcg caaatctggc tggttcaccc
     8461 gctattaaga agggtattct ccagaccgtg aaagtcgtgg acgagctggt caaggtgatg
     8521 ggtcgccata aaccagagaa cattgtcatc gagatggcca gggaaaacca gactacccag
     8581 aagggacaga agaacagcag ggagcggatg aaaagaattg aggaagggat taaggagctc
     8641 gggtcacaga tccttaaaga gcacccggtg gaaaacaccc agcttcagaa tgagaagctc
     8701 tatctgtact accttcaaaa tggacgcgat atgtatgtgg accaagagct tgatatcaac
     8761 aggctctcag actacgacgt ggacgccatc gtccctcaga gcttcctcaa agacgactca
     8821 attgacaata aggtgctgac tcgctcagac aagaaccggg gaaagtcaga taacgtgccc
     8881 tcagaggaag tcgtgaaaaa gatgaagaac tattggcgcc agcttctgaa cgcaaagctg
     8941 atcactcagc ggaagttcga caatctcact aaggctgaga ggggcggact gagcgaactg
     9001 gacaaagcag gattcattaa acggcaactt gtggagactc ggcagattac taaacatgtc
     9061 gcccaaatcc ttgactcacg catgaatacc aagtacgacg aaaacgacaa acttatccgc
     9121 gaggtgaagg tgattaccct gaagtccaag ctggtcagcg atttcagaaa ggactttcaa
     9181 ttctacaaag tgcgggagat caataactat catcatgctc atgacgcata tctgaatgcc
     9241 gtggtgggaa ccgccctgat caagaagtac ccaaagctgg aaagcgagtt cgtgtacgga
     9301 gactacaagg tctacgacgt gcgcaagatg attgccaaat ctgagcagga gatcggaaag
     9361 gccaccgcaa agtacttctt ctacagcaac atcatgaatt tcttcaagac cgaaatcacc
     9421 cttgcaaacg gtgagatccg gaagaggccg ctcatcgaga ctaatgggga gactggcgaa
     9481 atcgtgtggg acaagggcag agatttcgct accgtgcgca aagtgctttc tatgcctcaa
     9541 gtgaacatcg tgaagaaaac cgaggtgcaa accggaggct tttctaagga atcaatcctc
     9601 cccaagcgca actccgacaa gctcattgca aggaagaagg attgggaccc taagaagtac
     9661 ggcggattcg attcaccaac tgtggcttat tctgtcctgg tcgtggctaa ggtggaaaaa
     9721 ggaaagtcta agaagctcaa gagcgtgaag gaactgctgg gtatcaccat tatggagcgc
     9781 agctccttcg agaagaaccc aattgacttt ctcgaagcca aaggttacaa ggaagtcaag
     9841 aaggacctta tcatcaagct cccaaagtat agcctgttcg aactggagaa tgggcggaag
     9901 cggatgctcg cctccgctgg cgaacttcag aagggtaatg agctggctct cccctccaag
     9961 tacgtgaatt tcctctacct tgcaagccat tacgagaagc tgaaggggag ccccgaggac
    10021 aacgagcaaa agcaactgtt tgtggagcag cataagcatt atctggacga gatcattgag
    10081 cagatttccg agttttctaa acgcgtcatt ctcgctgatg ccaacctcga taaagtcctt
    10141 agcgcataca ataagcacag agacaaacca attcgggagc aggctgagaa tatcatccac
    10201 ctgttcaccc tcaccaatct tggtgcccct gccgcattca agtacttcga caccaccatc
    10261 gaccggaaac gctatacctc caccaaagaa gtgctggacg ccaccctcat ccaccagagc
    10321 atcaccggac tttacgaaac tcggattgac ctctcacagc tcggagggga tggtggcgga
    10381 ggctcgccaa aaaagaagag aaaggtagac ccaaagaaaa aacgaaaagt agatccgaaa
    10441 aagaagagga aggtggctag cgagggcaga ggaagtcttc taacatgcgg tgacgtggag
    10501 gagaatcccg gccctggtac catggccaag cctttgtctc aagaagaatc caccctcatt
    10561 gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag cgtcgccagc
    10621 gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca ttttactggg
    10681 ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc tggcaacctg
    10741 acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg cggacggtgc
    10801 cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga cagtgatgga
    10861 cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg ggagggcctg
    10921 cagctgcagt agttgataat agataacttc gtatagcata cattatacga agttatgaat
    10981 tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
    11041 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
    11101 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
    11161 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
    11221 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
    11281 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
    11341 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
    11401 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
    11461 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
    11521 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
    11581 cctccccgca tcgataccgt cgacctcgag acctagaaaa acatggagca atcacaagta
    11641 gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg
    11701 tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag
    11761 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
    11821 gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct gattggcaga
    11881 actacacacc agggccaggg atcagatatc cactgacctt tggatggtgc tacaagctag
    11941 taccagttga gcaagagaag gtagaagaag ccaatgaagg agagaacacc cgcttgttac
    12001 accctgtgag cctgcatggg atggatgacc cggagagaga agtattagag tggaggtttg
    12061 acagccgcct agcatttcat cacatggccc gagagctgca tccggactgt actgggtctc
    12121 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
    12181 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
    12241 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagggccc
    12301 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
    12361 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
    12421 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
    12481 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
    12541 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
    12601 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
    12661 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
    12721 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
    12781 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
    12841 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
    12901 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
    12961 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
    13021 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
    13081 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
    13141 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
    13201 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
    13261 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
    13321 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
    13381 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
    13441 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc
    13501 cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct
    13561 cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac
    13621 cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg
    13681 ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg
    13741 ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc
    13801 cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt
    13861 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
    13921 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
    13981 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
    14041 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
    14101 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
    14161 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
    14221 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
    14281 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
    14341 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
    14401 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
    14461 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
    14521 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
    14581 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
    14641 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
    14701 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
    14761 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
    14821 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
    14881 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
    14941 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
    15001 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
    15061 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
    15121 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
    15181 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
    15241 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
    15301 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
    15361 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
    15421 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
    15481 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
    15541 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
    15601 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
    15661 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
    15721 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
    15781 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
    15841 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
    15901 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
    15961 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
    16021 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
    16081 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
    16141 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
    16201 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    16261 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgac
//
