LOCUS       sequence-235431-       14933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of AB1-fused-dcas9 plasmids.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14933)
  AUTHORS   Wang H, Xu X, Nguyen CM, Liu Y, Gao Y, Lin X, Daley T, Kipniss NH, 
            La Russa M, Qi LS
  TITLE     CRISPR-Mediated Programmable 3D Genome Positioning and Nuclear 
            Organization.
  JOURNAL   Cell. 2018 Nov 15;175(5):1405-1417.e14. doi: 
            10.1016/j.cell.2018.09.013. Epub 2018 Oct 11.
  PUBMED    30318144
REFERENCE   2  (bases 1 to 14933)
  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"; doi: 
            "10.1016/j.cell.2018.09.013"; journalName: "Cell"; date: 
            "2018-11-15- 15"; volume: "175"; issue: "5"; pages: "1405-1417.e14"
COMMENT     Sequence Label: pHR-PGK-ABI-dCas9-P2A-mcherry
FEATURES             Location/Qualifiers
     source          1..14933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     95..112
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(182..202)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        187..516
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      367..502
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          1650..1715
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             1845..1865
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2290..2424
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2327..2346)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2381..2400
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             2592..3225
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    3272..3397
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3894..4127
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4312..4356
                     /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    4641..4758
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4823..5322
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4858..4881)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5255..5274
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             join(6313..6339,6340..6342,6343..8829,8830..8832,
                     8833..10416)
                     /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             6313..10416
                     /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             10420..10446
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             10426..10452
                     /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             10465..10485
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10492..10512
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10561..10617
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             10618..11328
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(10763..10781)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     11031..11050
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    11388..11976
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11441..11461)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11859..11870)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(11979..11995)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11980..11996)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             12086..12319
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(12344..12362)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(12345..12362)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(12652..12671)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(12885..12904)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     13004..13026
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(13064..13082)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        13150..13254
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(13255..13323,13324..14115)
                     /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     complement(13473..13492)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      14286..14874
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     14775..14794
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
       61 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
      121 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
      181 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
      241 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
      301 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
      361 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
      421 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
      481 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttggacacaa gacaggcttg
      541 cgagatatgt ttgagaatac cactttatcc cgcgtcaggg agaggcagtg cgtaaaaaga
      601 cgcggactca tgtgaaatac tggtttttag tgcgccagat ctctataatc tcgcgcaacc
      661 tattttcccc tcgaacactt tttaagccgt agataaacag gctgggacac ttcacatgag
      721 cgaaaaatac atcgtcacct gggacatgtt gcagatccat gcacgtaaac tcgcaagccg
      781 actgatgcct tctgaacaat ggaaaggcat tattgccgta agccgtggcg gtctgtaccg
      841 ggtgcgttac tggcgcgtga actgggtatt cgtcatgtcg ataccgtttg tatttccagc
      901 tacgatcacg acaaccagcg cgagcttaaa gtgctgaaac gcgcagaagg cgatggcgaa
      961 ggcttcatcg ttattgatga cctggtggat accggtggta ctgcggttgc gattcgtgaa
     1021 atgtatccaa aagcgcactt tgtcaccatc ttcgcaaaac cggctggtcg tccgctggtt
     1081 gatgactatg ttgttgatat cccgcaagat acctggattg aacagccgtg ggatatgggc
     1141 gtcgtattcg tcccgccaat ctccggtcgc taatcttttc aacgcctggc actgccgggc
     1201 gttgttcttt ttaacttcag gcgggttaca atagtttcca gtaagtattc tggaggctgc
     1261 atccatgaca caggcaaacc tgagcgaaac cctgttcaaa ccccgcttta aacatcctga
     1321 aacctcgacg ctagtccgcc gctttaatca cggcgcacaa ccgcctgtgc agtcggccct
     1381 tgatggtaaa accatccctc actggtatcg catgattaac cgtctgatgt ggatctggcg
     1441 cggcattgac ccacgcgaaa tcctcgacgt ccaggcacgt attgtgatga gcgatgccga
     1501 acgtaccgac gatgatttat acgatacggt gattggctac cgtggcggca actggattta
     1561 tgagtgggcc ccggatcttt gtgaaggaac cttacttctg tggtgtgaca taattggaca
     1621 aactacctac agagatttaa agctctaagg taaatataaa atttttaagt gtataatgtg
     1681 ttaaactact gattctaatt gtttgtgtat tttagattcc aacctatgga actgatgaat
     1741 gggagcagtg gtggaatgcc tttaatgagg aaaacctgtt ttgctcagaa gaaatgccat
     1801 ctagtgatga tgaggctact gctgactctc aacattctac tcctccaaaa aagaagagaa
     1861 aggtagaaga ccccaaggac tttccttcag aattgctaag ttttttgagt catgctgtgt
     1921 ttagtaatag aactcttgct tgctttgcta tttacaccac aaaggaaaaa gctgcactgc
     1981 tatacaagaa aattatggaa aaatattctg taacctttat aagtaggcat aacagttata
     2041 atcataacat actgtttttt cttactccac acaggcatag agtgtctgct attaataact
     2101 atgctcaaaa attgtgtacc tttagctttt taatttgtaa aggggttaat aaggaatatt
     2161 tgatgtatag tgccttgact agagatcata atcagccata ccacatttgt agaggtttta
     2221 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     2281 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2341 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2401 aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa
     2461 cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc
     2521 tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca
     2581 aacttagtag ttggaagggc taattcactc ccaaagaaga caagatatcc ttgatctgtg
     2641 gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt
     2701 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt
     2761 agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat
     2821 ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca
     2881 cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa
     2941 gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg
     3001 agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag
     3061 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     3121 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     3181 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag
     3241 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga
     3301 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
     3361 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
     3421 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
     3481 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
     3541 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
     3601 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
     3661 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
     3721 caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga tatgagggac
     3781 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     3841 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     3901 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     3961 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     4021 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     4081 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     4141 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     4201 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     4261 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     4321 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     4381 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     4441 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     4501 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
     4561 ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg gtatcgccaa
     4621 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     4681 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     4741 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttggatcgat
     4801 aagcttgata tcgaattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca
     4861 tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca
     4921 cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc cttcgcgcca
     4981 ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg
     5041 acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag
     5101 caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc
     5161 tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc
     5221 ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg
     5281 tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag cccaagctta
     5341 cgcgtctgca ggatatcaag cttgcggtac cgcgggcccg gggccaccat gacgcgtgtg
     5401 cctttgtatg gttttacttc gatttgtgga agaagacctg agatggaagc tgctgtttcg
     5461 actataccaa gattccttca atcttcctct ggttcgatgt tagatggtcg gtttgatcct
     5521 caatccgccg ctcatttctt cggtgtttac gacggccatg gcggttctca ggtagcgaac
     5581 tattgtagag agaggatgca tttggctttg gcggaggaga tagctaagga gaaaccgatg
     5641 ctctgcgatg gtgatacgtg gctggagaag tggaagaaag ctcttttcaa ctcgttcctg
     5701 agagttgact cggagattga gtcagttgcg ccggagacgg ttgggtcaac gtcggtggtt
     5761 gccgttgttt tcccgtctca catcttcgtc gctaactgcg gtgactctag agccgttctt
     5821 tgccgcggca aaactgcact tccattatcc gttgaccata aaccggatag agaagatgaa
     5881 gctgcgagga ttgaagccgc aggagggaaa gtgattcagt ggaatggagc tcgtgttttc
     5941 ggtgttctcg ccatgtcgag atccattggc gatagatact tgaaaccatc catcattcct
     6001 gatccggaag tgacggctgt gaagagagta aaagaagatg attgtctgat tttggcgagt
     6061 gacggggttt gggatgtaat gacggatgaa gaagcgtgtg agatggcaag gaagcggatt
     6121 ctcttgtggc acaagaaaaa cgcggtggct ggggatgcat cgttgctcgc ggatgagcgg
     6181 agaaaggaag ggaaagatcc tgcggcgatg tccgcggctg agtatttgtc aaagctggcg
     6241 atacagagag gaagcaaaga caacataagt gtggtggtgg ttgatttgaa gcatatgggt
     6301 ggaggctccg ggatggacaa gaagtacagc atcggcctgg ccatcggcac caactctgtg
     6361 ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc
     6421 aacaccgacc ggcacagcat caagaagaac ctgatcggcg ccctgctgtt cgacagcgga
     6481 gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag
     6541 aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc
     6601 ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac
     6661 cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac
     6721 cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg
     6781 gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc
     6841 gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc
     6901 gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg
     6961 agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc
     7021 ctgttcggca acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc
     7081 gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac
     7141 aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg
     7201 tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc
     7261 ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa
     7321 gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag
     7381 aacggctacg ccggctacat cgatggcgga gccagccagg aagagttcta caagttcatc
     7441 aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag
     7501 gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg
     7561 ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac
     7621 cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc
     7681 aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg
     7741 aacttcgagg aagtggtgga caagggcgcc agcgcccaga gcttcatcga gcggatgacc
     7801 aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag
     7861 tacttcaccg tgtacaacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag
     7921 cccgccttcc tgagcggcga gcagaaaaaa gccatcgtgg acctgctgtt caagaccaac
     7981 cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac
     8041 tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat
     8101 ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg
     8161 gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg
     8221 aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac
     8281 accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc
     8341 aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg
     8401 atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag
     8461 ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc
     8521 atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc
     8581 gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac
     8641 agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg
     8701 aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg
     8761 cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac
     8821 gatgtggacg ctatcgtgcc tcagagcttt ctgaaggacg actccatcga taacaaagtg
     8881 ctgactcgga gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg
     8941 aagaagatga agaactactg gcgccagctg ctgaatgcca agctgattac ccagaggaag
     9001 ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc
     9061 atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac
     9121 tcccggatga acactaagta cgacgagaac gacaaactga tccgggaagt gaaagtgatc
     9181 accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc
     9241 gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc
     9301 ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac
     9361 gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac
     9421 ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag
     9481 atccggaagc ggcctctgat cgagacaaac ggcgaaacag gcgagatcgt gtgggataag
     9541 ggccgggact ttgccaccgt gcggaaagtg ctgtctatgc cccaagtgaa tatcgtgaaa
     9601 aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc
     9661 gacaagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc
     9721 cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa
     9781 ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag
     9841 aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc
     9901 aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct
     9961 gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg
    10021 tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag
    10081 ctgtttgtgg aacagcacaa acactacctg gacgagatca tcgagcagat cagcgagttc
    10141 tccaagagag tgatcctggc cgacgctaat ctggacaagg tgctgagcgc ctacaacaag
    10201 cacagagaca agcctatcag agagcaggcc gagaatatca tccacctgtt taccctgacc
    10261 aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac
    10321 accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac
    10381 gagacacgga tcgacctgtc tcagctggga ggcgacgcct atccctatga cgtgcccgat
    10441 tatgccagcc tgggcagcgg ctcccccaag aaaaaacgca aggtggaaga tcctaagaaa
    10501 aagcggaaag tggacggcat tggtagtggg agcaacggca gcagcggatc cggaagcgga
    10561 gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctatg
    10621 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
    10681 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
    10741 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
    10801 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
    10861 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
    10921 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
    10981 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
    11041 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
    11101 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
    11161 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
    11221 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
    11281 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaagc ggccgcgact
    11341 ctagagtcga cctgcaggca tgcaagcttg atatcaagct tatcgataat caacctctgg
    11401 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
    11461 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
    11521 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
    11581 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
    11641 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
    11701 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
    11761 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
    11821 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
    11881 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
    11941 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgaggga
    12001 attaattcga gctcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
    12061 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
    12121 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
    12181 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
    12241 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
    12301 gtgtggaaaa tctctagcag catctagaat taattccgtg tattctatag tgtcacctaa
    12361 atcgtatgtg tatgatacat aaggttatgt attaattgta gccgcgttct aacgacaata
    12421 tgtacaagcc taattgtgta gcatctggct tactgaagca gaccctatca tctctctcgt
    12481 aaactgccgt cagagtcggt ttggttggac gaaccttctg agtttctggt aacgccgtcc
    12541 cgcacccgga aatggtcagc gaaccaatca gcagggtcat cgctagccag atcctctacg
    12601 ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg
    12661 ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg
    12721 gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg
    12781 caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa
    12841 tgcaggagtc gcataaggga gagcgtcgaa tggtgcactc tcagtacaat ctgctctgat
    12901 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
    12961 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
    13021 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
    13081 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
    13141 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
    13201 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
    13261 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
    13321 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
    13381 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
    13441 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
    13501 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
    13561 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
    13621 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
    13681 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
    13741 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
    13801 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
    13861 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
    13921 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
    13981 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
    14041 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
    14101 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
    14161 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
    14221 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
    14281 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
    14341 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
    14401 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
    14461 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
    14521 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
    14581 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
    14641 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
    14701 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
    14761 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
    14821 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
    14881 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca cat
//
