LOCUS       sequence-171198-       14617 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing a KRAB-dCas9 fusion protein and BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14617)
  AUTHORS   Fulco CP, Munschauer M, Anyoha R, Munson G, Grossman SR, Perez EM, 
            Kane M, Cleary B, Lander ES, Engreitz JM
  TITLE     Systematic mapping of functional enhancer-promoter connections with 
            CRISPR interference.
  JOURNAL   Science. 2016 Sep 29. pii: aag2445.
  PUBMED    27708057
REFERENCE   2  (bases 1 to 14617)
  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: "Science. 
            2016 Sep 29. pii: aag2445."
COMMENT     Sequence Label: TRE-KRAB-dCas9-IRES-BFP
FEATURES             Location/Qualifiers
     source          1..14617
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(21..40)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      834..1422
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1323..1342
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1576..1593
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1663..1683)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1668..1997
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1848..1983
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          3131..3196
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3326..3346
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3771..3905
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3808..3827)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3862..3881
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             4073..4706
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    4753..4878
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5375..5608
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5793..5837
                     /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    6122..6239
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6301..6650
                     /label=TRE3GV promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, optimized for 
                     retroviral and lentiviral vectors"
     protein_bind    6309..6327
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6345..6363
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6381..6399
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6417..6435
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6453..6471
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6489..6507
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6525..6543
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     6589..6613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             6732..6917
                     /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             join(6939..6965,6966..6968,6969..9455,9456..9458,
                     9459..11042)
                     /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             6939..11042
                     /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
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             11046..11072
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             11052..11078
                     /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             11091..11111
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11118..11138
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    join(11221..11795,11796..11798,11799..11807)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(11388..11405)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     11615..11634
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             11808..12509
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    12553..13141
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(12606..12626)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(13024..13035)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(13144..13160)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13145..13161)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             13251..13484
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(13509..13527)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(13510..13527)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(13817..13836)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(14050..14069)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     14169..14191
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(14229..14247)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        14315..14419
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(14420..14488,14489..14617,1..663)
                     /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"
ORIGIN
        1 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
       61 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
      121 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
      181 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
      241 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
      301 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
      361 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
      421 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
      481 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
      541 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
      601 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
      661 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
      721 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
      781 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
      841 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
      901 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
      961 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     1021 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1081 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     1141 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     1201 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     1261 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     1321 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     1381 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     1441 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     1501 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     1561 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     1621 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct gtggaatgtg tgtcagttag
     1681 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     1741 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     1801 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     1861 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     1921 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     1981 gcctaggctt ttgcaaaaag cttggacaca agacaggctt gcgagatatg tttgagaata
     2041 ccactttatc ccgcgtcagg gagaggcagt gcgtaaaaag acgcggactc atgtgaaata
     2101 ctggttttta gtgcgccaga tctctataat ctcgcgcaac ctattttccc ctcgaacact
     2161 ttttaagccg tagataaaca ggctgggaca cttcacatga gcgaaaaata catcgtcacc
     2221 tgggacatgt tgcagatcca tgcacgtaaa ctcgcaagcc gactgatgcc ttctgaacaa
     2281 tggaaaggca ttattgccgt aagccgtggc ggtctgtacc gggtgcgtta ctggcgcgtg
     2341 aactgggtat tcgtcatgtc gataccgttt gtatttccag ctacgatcac gacaaccagc
     2401 gcgagcttaa agtgctgaaa cgcgcagaag gcgatggcga aggcttcatc gttattgatg
     2461 acctggtgga taccggtggt actgcggttg cgattcgtga aatgtatcca aaagcgcact
     2521 ttgtcaccat cttcgcaaaa ccggctggtc gtccgctggt tgatgactat gttgttgata
     2581 tcccgcaaga tacctggatt gaacagccgt gggatatggg cgtcgtattc gtcccgccaa
     2641 tctccggtcg ctaatctttt caacgcctgg cactgccggg cgttgttctt tttaacttca
     2701 ggcgggttac aatagtttcc agtaagtatt ctggaggctg catccatgac acaggcaaac
     2761 ctgagcgaaa ccctgttcaa accccgcttt aaacatcctg aaacctcgac gctagtccgc
     2821 cgctttaatc acggcgcaca accgcctgtg cagtcggccc ttgatggtaa aaccatccct
     2881 cactggtatc gcatgattaa ccgtctgatg tggatctggc gcggcattga cccacgcgaa
     2941 atcctcgacg tccaggcacg tattgtgatg agcgatgccg aacgtaccga cgatgattta
     3001 tacgatacgg tgattggcta ccgtggcggc aactggattt atgagtgggc cccggatctt
     3061 tgtgaaggaa ccttacttct gtggtgtgac ataattggac aaactaccta cagagattta
     3121 aagctctaag gtaaatataa aatttttaag tgtataatgt gttaaactac tgattctaat
     3181 tgtttgtgta ttttagattc caacctatgg aactgatgaa tgggagcagt ggtggaatgc
     3241 ctttaatgag gaaaacctgt tttgctcaga agaaatgcca tctagtgatg atgaggctac
     3301 tgctgactct caacattcta ctcctccaaa aaagaagaga aaggtagaag accccaagga
     3361 ctttccttca gaattgctaa gttttttgag tcatgctgtg tttagtaata gaactcttgc
     3421 ttgctttgct atttacacca caaaggaaaa agctgcactg ctatacaaga aaattatgga
     3481 aaaatattct gtaaccttta taagtaggca taacagttat aatcataaca tactgttttt
     3541 tcttactcca cacaggcata gagtgtctgc tattaataac tatgctcaaa aattgtgtac
     3601 ctttagcttt ttaatttgta aaggggttaa taaggaatat ttgatgtata gtgccttgac
     3661 tagagatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     3721 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     3781 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3841 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     3901 ggatcaactg gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta
     3961 ttaccactgc caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat
     4021 tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta gttggaaggg
     4081 ctaattcact cccaaagaag acaagatatc cttgatctgt ggatctacca cacacaaggc
     4141 tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc actgaccttt
     4201 ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc caataaagga
     4261 gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa
     4321 gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat
     4381 ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc cgctggggac
     4441 tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata
     4501 taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga
     4561 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     4621 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     4681 ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg
     4741 aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc
     4801 gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga
     4861 gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt
     4921 cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg
     4981 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
     5041 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
     5101 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
     5161 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
     5221 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     5281 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     5341 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     5401 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     5461 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     5521 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     5581 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     5641 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     5701 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     5761 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     5821 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     5881 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     5941 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     6001 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     6061 gatccattcg attagtgaac ggatctcgac ggtatcgcca aatggcagta ttcatccaca
     6121 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     6181 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc
     6241 gggtttatta cagggacagc agagatccag tttatcgatg aggccctttc gtcttcactc
     6301 gagtttactc cctatcagtg atagagaacg tatgaagagt ttactcccta tcagtgatag
     6361 agaacgtatg cagactttac tccctatcag tgatagagaa cgtataagga gtttactccc
     6421 tatcagtgat agagaacgta tgaccagttt actccctatc agtgatagag aacgtatcta
     6481 cagtttactc cctatcagtg atagagaacg tatatccagt ttactcccta tcagtgatag
     6541 agaacgtatg tcgaggtagg cgtgtacggt gggcgcctat aaaagcagag ctcgtttagt
     6601 gaaccgtcag atcgcctgga gcaattccac aacacttttg tcttatactt acgcgtctgc
     6661 aggatatcaa gcttgcggta ccgcgggccc gggatcgcac catggatgct aagtcactaa
     6721 ctgcctggtc ccggacactg gtgaccttca aggatgtatt tgtggacttc accagggagg
     6781 agtggaagct gctggacact gctcagcaga tcgtgtacag aaatgtgatg ctggagaact
     6841 ataagaacct ggtttccttg ggttatcagc ttactaagcc agatgtgatc ctccggttgg
     6901 agaagggaga agagcccggc ggttccggcg gagggtcgat ggacaagaag tacagcatcg
     6961 gcctggccat cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc
     7021 ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga
     7081 tcggcgccct gctgttcgac agcggagaaa cagccgaggc cacccggctg aagagaaccg
     7141 ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca
     7201 acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg
     7261 aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct
     7321 accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca
     7381 aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact
     7441 tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc
     7501 tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg
     7561 ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc
     7621 agctgcccgg cgagaagaag aatggcctgt tcggcaacct gattgccctg agcctgggcc
     7681 tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca
     7741 aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg
     7801 acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag
     7861 tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc
     7921 accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca
     7981 aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacatcgat ggcggagcca
     8041 gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg
     8101 aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg
     8161 gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag
     8221 atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca
     8281 tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa
     8341 agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgccagcg
     8401 cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc
     8461 tgcccaagca cagcctgctg tacgagtact tcaccgtgta caacgagctg accaaagtga
     8521 aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaagcca
     8581 tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact
     8641 acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca
     8701 acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg
     8761 acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg
     8821 acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga
     8881 tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca
     8941 acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct
     9001 tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca
     9061 tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg
     9121 ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg
     9181 tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc
     9241 agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca
     9301 tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga
     9361 acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac
     9421 tggacatcaa ccggctgtcc gactacgatg tggacgctat cgtgcctcag agctttctga
     9481 aggacgactc catcgataac aaagtgctga ctcggagcga caagaaccgg ggcaagagcg
     9541 acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgc cagctgctga
     9601 atgccaagct gattacccag aggaagttcg acaatctgac caaggccgag agaggcggcc
     9661 tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca
     9721 caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac gagaacgaca
     9781 aactgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga
     9841 aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct
     9901 acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt
     9961 tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg
    10021 aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga
    10081 ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg
    10141 aaacaggcga gatcgtgtgg gataagggcc gggactttgc caccgtgcgg aaagtgctgt
    10201 ctatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag
    10261 agtctatcct gcccaagagg aacagcgaca agctgatcgc cagaaagaag gactgggacc
    10321 ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca
    10381 aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca
    10441 tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca
    10501 aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa
    10561 acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc
    10621 tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct
    10681 cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaaacac tacctggacg
    10741 agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg
    10801 acaaggtgct gagcgcctac aacaagcaca gagacaagcc tatcagagag caggccgaga
    10861 atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg
    10921 acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga
    10981 tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggcg
    11041 acgcctatcc ctatgacgtg cccgattatg ccagcctggg cagcggctcc cccaagaaaa
    11101 aacgcaaggt ggaagatcct aagaaaaagc ggaaagtgga cggcattggt agtgggagca
    11161 acggcagcag cggatcctaa gcggccgctc gataagcttg atatcgaatt ccgccccccc
    11221 cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
    11281 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
    11341 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
    11401 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
    11461 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
    11521 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
    11581 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
    11641 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
    11701 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
    11761 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg agcgagctga
    11821 ttaaggagaa catgcacatg aagctgtaca tggagggcac cgtggacaac catcacttca
    11881 agtgcacatc cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg
    11941 tggtcgaggg cggccctctc cccttcgcct tcgacatcct ggctactagc ttcctctacg
    12001 gcagcaagac cttcatcaac cacacccagg gcatccccga cttcttcaag cagtccttcc
    12061 ctgagggctt cacatgggag agagtcacca catacgaaga cgggggcgtg ctgaccgcta
    12121 cccaggacac cagcctccag gacggctgcc tcatctacaa cgtcaagatc agaggggtga
    12181 acttcacatc caacggccct gtgatgcaga agaaaacact cggctgggag gccttcaccg
    12241 agacgctgta ccccgctgac ggcggcctgg aaggcagaaa cgacatggcc ctgaagctcg
    12301 tgggcgggag ccatctgatc gcaaacatca agaccacata tagatccaag aaacccgcta
    12361 agaacctcaa gatgcctggc gtctactatg tggactacag actggaaaga atcaaggagg
    12421 ccaacaacga gacctacgtc gagcagcacg aggtggcagt ggccagatac tgcgacctcc
    12481 ctagcaaact ggggcacaag cttaattaaa ctagtcctgc aggcatgcaa gcttgatatc
    12541 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
    12601 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
    12661 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
    12721 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
    12781 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
    12841 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
    12901 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
    12961 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
    13021 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
    13081 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
    13141 catcgatacc gtcgacctcg agggaattaa ttcgagctcg gtacctttaa gaccaatgac
    13201 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
    13261 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
    13321 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
    13381 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
    13441 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagcatct agaattaatt
    13501 ccgtgtattc tatagtgtca cctaaatcgt atgtgtatga tacataaggt tatgtattaa
    13561 ttgtagccgc gttctaacga caatatgtac aagcctaatt gtgtagcatc tggcttactg
    13621 aagcagaccc tatcatctct ctcgtaaact gccgtcagag tcggtttggt tggacgaacc
    13681 ttctgagttt ctggtaacgc cgtcccgcac ccggaaatgg tcagcgaacc aatcagcagg
    13741 gtcatcgcta gccagatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca
    13801 ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac
    13861 ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga
    13921 ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc
    13981 aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgaatggtg
    14041 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
    14101 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
    14161 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
    14221 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
    14281 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
    14341 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
    14401 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
    14461 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
    14521 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
    14581 ccttgagagt tttcgccccg aagaacgttt tccaatg
//
