LOCUS       sequence-239644-       15284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Catalytically inactive Cas9 from S. pyogenes with P2A-EGFP under the
            EF1a core promoter, and cloning backbone for sgRNA. Contains BsmBI 
            sites for insertion of spacer sequences..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15284)
  TITLE     Ferrer Lab Plasmids
REFERENCE   2  (bases 1 to 15284)
  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"
COMMENT     Sequence Label: Lenti-(BB)-EF1a-KRAB-dCas9-P2A-EGFP
FEATURES             Location/Qualifiers
     source          1..15284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(176..280)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(355..374)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        546..925
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        927..1125
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1080..1100
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1143..1323
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1370..1495
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1988..2221
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2406..2450
                     /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    2748..2865
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2916..3164
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2916..2936
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3087..3106
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        5046..5121
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        5183..5394
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5407..5416
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5443..5628
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             5650..9753
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTSLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             9757..9783
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             9763..9789
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             9802..9822
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9829..9849
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9892..9945
                     /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(9946..9948,9949..10659)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(9946..9948,9949..10659)
                     /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(10249..10268)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     10596..10617
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    10684..11272
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10737..10757)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11155..11166)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11344..11577
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(11603..11620)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11609..11833
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11879..12307
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11966..11985)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12176..12197
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12316..12336)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12321..12650
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12501..12636
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12563..12582
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12698..12745
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12764..13138
                     /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    13268..13389
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13305..13324)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13359..13378
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13438..13454)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13438..13454)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13451..13473)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13462..13478
                     /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(13486..13492,13493..13510,13511..13516))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13531..13552
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13669..13686)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13840..14428)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13920..13939)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14599..15284,1..106,107..175))
                     /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     15222..15241
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
       61 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
      121 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
      181 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
      241 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
      301 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
      361 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
      421 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
      481 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
      541 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      601 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      661 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      721 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      781 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      841 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      901 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      961 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     1021 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     1081 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagcgcgt tttgcctgta
     1141 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     1201 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     1261 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     1321 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg
     1381 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
     1441 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
     1501 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
     1561 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
     1621 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
     1681 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
     1741 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga
     1801 gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag
     1861 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     1921 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     1981 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     2041 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     2101 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     2161 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     2221 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     2281 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     2341 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     2401 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     2461 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     2521 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     2581 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa
     2641 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg
     2701 cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa
     2761 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
     2821 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
     2881 gacagcagag atccagtttg gttaattaag gtaccgaggg cctatttccc atgattcctt
     2941 catatttgca tatacgatac aaggctgtta gagagataat tagaattaat ttgactgtaa
     3001 acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg
     3061 cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt
     3121 tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccggagac ggttgtaaat
     3181 gagcacacaa aatacacatg ctaaaatatt atattctatg acctttataa aatcaaccaa
     3241 aatcttcttt ttaataactt tagtatcaat aattagaatt tttatgttcc tttttgcaaa
     3301 cttttaataa aaatgagcaa aataaaaaaa cgctagtttt agtaactcgc gttgttttct
     3361 tcacctttaa taatagctac tccaccactt gttcctaagc ggtcagctcc tgcttcaatc
     3421 attttttgag catcttcaaa tgttctaact ccaccagctg ctttaactaa agcattgtct
     3481 ttaacaactg acttcattag tttaacatct tcaaatgttg cacctgattt tgaaaatcct
     3541 gttgatgttt taacaaattc taatccagct tcaacagcta tttcacaagc tttcatgatt
     3601 tcttcttttg ttaataaaca attttccata atacatttaa caacatgtga tccagctgct
     3661 ttttttacag ctttcatgtc ttctaaaact aattcataat ttttgtcttt taatgcacca
     3721 atatttaata ccatatcaat ttctgttgca ccatctttaa ttgcttcaga aacttcgaat
     3781 gcttttgtag ctgttgtgca tgcacctaga ggaaaaccta caacatttgt tattcctaca
     3841 tttgtgcctt ttaataattc tttacaatag cttgttcaat atgaattaac acaaactgtt
     3901 gcaaaatcaa attcaattgc ttcatcacat aattgtttaa tttcagcttt cgtagcatct
     3961 tgttttaata atgtgtgatc tatatatttg tttagtttca ttttttctcc tatatattca
     4021 tttttaattt taattcttta ataatttcgt ctactttaac tttagcgttt tgaacagatt
     4081 caccaacacc tataaaataa atttttagtt taggttcagt tccacttggg cgaacagcaa
     4141 atcatgactt atcttctaaa taaaatttta gtaagtcttg tcctggcata ttatacattc
     4201 catcgatgta gtcttcaaca ttaacaactt taagtccagc aatttgagtt aagggtgttg
     4261 ctctcaatga tttcattaat ggttcaattt ttaatttctt ttcttctggt ttaaaattca
     4321 agtttaaagt gaaagtgtaa tatgcaccca tttctttaaa taaatcttct aaatagtcta
     4381 ctaatgtttt attttgtttt ttataaaatc aagcagcctc tgctattaat atagaagctt
     4441 gtattccatc tttatctcta gctgagtcat caattacata tccataactt tcttcataag
     4501 caaaaacaaa atttaatccg ttatcttctt ctttagcaat ttctctaccc attcatttaa
     4561 atccagttaa agtttttaca atattaactc catatttttc atgagcgatt ctatcaccca
     4621 aatcacttgt tacaaaactt gaatatagag ccggattttt tggaatgcta tttaagcgtt
     4681 ttagatttga taattttcaa tcaattaaaa ttggtcctgt ttgatttcca tctaatctta
     4741 caaaatgacc atcatgtttt attgccattc caaatctgtc agcatctggg tcattcataa
     4801 taataatatc tgcatcatgt ttaataccat attcaagcgg tatttttcat gcaggatcaa
     4861 attctggatt tggatttaca acatttttaa atgtttcatc ttcaaatgca tgctcttcaa
     4921 cctcaataac gttatatcct gattcacgta atatttttgg ggtaaattta gttcctgttc
     4981 cattaactgc gctaaaaata atttttaaat cttttttagc ttcttgctct tttttgtacg
     5041 tctctgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
     5101 aaagtggcac cgagtcggtg cttttttgaa ttcgctagct aggtcttgaa aggagtggga
     5161 attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
     5221 gggggagggg tcggcaattg atccggtgcc tagagaaggt ggcgcggggt aaactgggaa
     5281 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
     5341 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggaccgg
     5401 tgatccgcca ccatggatgc taagtcacta actgcctggt cccggacact ggtgaccttc
     5461 aaggatgtat ttgtggactt caccagggag gagtggaagc tgctggacac tgctcagcag
     5521 atcgtgtaca gaaatgtgat gctggagaac tataagaacc tggtttcctt gggttatcag
     5581 cttactaagc cagatgtgat cctccggttg gagaagggag aagagcccgg cggttccggc
     5641 ggagggtcga tggacaagaa gtacagcatc ggcctggcca tcggcaccaa ctctgtgggc
     5701 tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac
     5761 accgaccggc acagcatcaa gaagaacctg atcggcgccc tgctgttcga cagcggagaa
     5821 acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac
     5881 cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc
     5941 ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc
     6001 atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac
     6061 ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc
     6121 ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac
     6181 aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag
     6241 gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc
     6301 aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg
     6361 ttcggcaacc tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac
     6421 ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac
     6481 ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc
     6541 gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg
     6601 agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct
     6661 ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac
     6721 ggctacgccg gctacatcga tggcggagcc agccaggaag agttctacaa gttcatcaag
     6781 cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac
     6841 ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga
     6901 gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg
     6961 gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg
     7021 ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac
     7081 ttcgaggaag tggtggacaa gggcgccagc gcccagagct tcatcgagcg gatgaccaac
     7141 ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac
     7201 ttcaccgtgt acaacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc
     7261 gccttcctga gcggcgagca gaaaaaagcc atcgtggacc tgctgttcaa gaccaaccgg
     7321 aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc
     7381 gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg
     7441 ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa
     7501 gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa
     7561 acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc
     7621 ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag
     7681 acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc
     7741 cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc
     7801 gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc
     7861 ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag
     7921 aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc
     7981 cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa
     8041 gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag
     8101 aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat
     8161 gtggacgcta tcgtgcctca gagctttctg aaggacgact ccatcgataa caaagtgctg
     8221 actcggagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag
     8281 aagatgaaga actactggcg ccagctgctg aatgccaagc tgattaccca gaggaagttc
     8341 gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc
     8401 aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc
     8461 cggatgaaca ctaagtacga cgagaacgac aaactgatcc gggaagtgaa agtgatcacc
     8521 ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag
     8581 atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg
     8641 atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac
     8701 gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc
     8761 ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc
     8821 cggaagcggc ctctgatcga gacaaacggc gaaacaggcg agatcgtgtg ggataagggc
     8881 cgggactttg ccaccgtgcg gaaagtgctg tctatgcccc aagtgaatat cgtgaaaaag
     8941 accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgac
     9001 aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc
     9061 accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg
     9121 aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat
     9181 cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag
     9241 ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc
     9301 ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac
     9361 ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg
     9421 tttgtggaac agcacaaaca ctacctggac gagatcatcg agcagatcag cgagttctcc
     9481 aagagagtga tcctggccga cgctaatctg gacaaggtgc tgagcgccta caacaagcac
     9541 agagacaagc ctatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccagt
     9601 ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc
     9661 agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag
     9721 acacggatcg acctgtctca gctgggaggc gacgcctatc cctatgacgt gcccgattat
     9781 gccagcctgg gcagcggctc ccccaagaaa aaacgcaagg tggaagatcc taagaaaaag
     9841 cggaaagtgg acggcattgg tagtgggagc aacggcagca gcggggatcc cgagggcaga
     9901 ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccagtgag caagggcgag
     9961 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
    10021 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
    10081 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
    10141 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
    10201 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
    10261 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
    10321 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
    10381 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
    10441 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
    10501 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
    10561 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
    10621 gccgccggga tcactctcgg catggacgag ctgtacaagg aattctaaac gcgttaagtc
    10681 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
    10741 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
    10801 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
    10861 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
    10921 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
    10981 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
    11041 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
    11101 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
    11161 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
    11221 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcgtcgactt
    11281 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
    11341 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
    11401 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
    11461 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
    11521 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaggg
    11581 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
    11641 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
    11701 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
    11761 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
    11821 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
    11881 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
    11941 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
    12001 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
    12061 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
    12121 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
    12181 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
    12241 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
    12301 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
    12361 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
    12421 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
    12481 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
    12541 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
    12601 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
    12661 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
    12721 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
    12781 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
    12841 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
    12901 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
    12961 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
    13021 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
    13081 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
    13141 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
    13201 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
    13261 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
    13321 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
    13381 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
    13441 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
    13501 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
    13561 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
    13621 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
    13681 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
    13741 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
    13801 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
    13861 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
    13921 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
    13981 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
    14041 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
    14101 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
    14161 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
    14221 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
    14281 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
    14341 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
    14401 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
    14461 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
    14521 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
    14581 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
    14641 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
    14701 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
    14761 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
    14821 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
    14881 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
    14941 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
    15001 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
    15061 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
    15121 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
    15181 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
    15241 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggc
//
