LOCUS       sequence-352468-       12279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ZIM3 KRAB domain fused to the N terminus of dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12279)
  AUTHORS   Alerasool N, Segal D, Lee H, Taipale M
  TITLE     An efficient KRAB domain for CRISPRi applications in human cells.
  JOURNAL   Nat Methods. 2020 Nov;17(11):1093-1096. doi: 
            10.1038/s41592-020-0966-x. Epub 2020 Oct 5.
  PUBMED    33020655
REFERENCE   2  (bases 1 to 12279)
  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: 
            "10.1038/s41592-020-0966-x"; journalName: "Nat Methods"; date: 
            "2020-11"; volume: "17"; issue: "11"; pages: "1093-1096"
COMMENT     Sequence Label: pLX303-ZIM3-KRAB-dCas9
FEATURES             Location/Qualifiers
     source          1..12279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(36..827,828..896))
                     /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     659..678
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(897..1001)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1027..1482)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1164..1185)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1376..1395
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1609..1631
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1623..1640
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1624..1640
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1650..1669
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1650..1668
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      1689..1824
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1864..1985
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1875..1894)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1929..1948
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             2063..2296
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    2368..2956
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2474..2485
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2883..2903
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3004..3024)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(3031..3051)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(3064..3090)
                     /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             complement(3070..3096)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(join(3100..4683,4684..4686,4687..7173,7174..7176,
                     7177..7203))
                     /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             complement(3100..7203)
                     /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"
     regulatory      7200..7209
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(7258..7278)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    7298..7322
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(7623..7647)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(7670..7873)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(7699..7719)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7874..8177
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    8294..8411
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(8471..8866)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(8878..9378)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8941..8959)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             complement(9553..9597)
                     /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    9782..10015
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    10508..10633
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             10680..10860
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(10861..11087)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(11115..11135)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(11115..11133)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11154..11170)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11154..11170)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11167..11189)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11178..11194
                     /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(11202..11208,11209..11226,11227..11232))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11247..11268
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11385..11402)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11556..12144)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11636..11655)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
       61 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
      121 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
      181 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
      241 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
      301 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
      361 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
      421 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
      481 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
      541 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
      601 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
      661 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
      721 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
      781 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
      841 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
      901 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
      961 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     1021 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     1081 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     1141 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     1201 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     1261 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     1321 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     1381 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     1441 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     1501 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     1561 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     1621 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta
     1681 cgtccctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggccgaggc
     1741 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     1801 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggatagcta gagccagaca
     1861 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     1921 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     1981 aagttcctct cactctctga tattcatttc tttgcaagtt ataaatactg aataataaga
     2041 tgacatgaac tactatacgt actgctagag attttccaca ctgactaaaa gggtctgagg
     2101 gatctctagt taccagagtc acacaacaga cgggcacaca ctacttgaag cactcaaggc
     2161 aagctttatt gaggcttaag cagtgggttc cctagttagc cagagagctc ccaggctcag
     2221 atctggtcta accagagaga cccagtacaa gcaaaaagca gatcttgtct tcgttgggag
     2281 tgaattagcc cttccagtcc ccccttttct tttaaaaagt ggctaagatc tacagctgcc
     2341 ttgtaagtca ttggtcttaa agtcgacgcg gggaggcggc ccaaagggag atccgactcg
     2401 tctgagggcg aaggcgaaga cgcggaagag gccgcagagc cggcagcagg ccgcgggaag
     2461 gaaggtccgc tggattgagg gccgaaggga cgtagcagaa ggacgtcccg cgcagaatcc
     2521 aggtggcaac acaggcgagc agccaaggaa aggacgatga tttccccgac aacaccacgg
     2581 aattgtcagt gcccaacagc cgagcccctg tccagcagcg ggcaaggcag gcggcgatga
     2641 gttccgccgt ggcaataggg agggggaaag cgaaagtccc ggaaaggagc tgacaggtgg
     2701 tggcaatgcc ccaaccagtg ggggttgcgt cagcaaacac agtgcacacc acgccacgtt
     2761 gcctgacaac gggccacaac tcctcataaa gagacagcaa ccaggattta tacaaggagg
     2821 agaaaatgaa agccatacgg gaagcaatag catgatacaa aggcattaaa gcagcgtatc
     2881 cacatagcgt aaaaggagca acatagttaa gaataccagt caatctttca caaattttgt
     2941 aatccagagg ttgattgtcg acttaacgcg ccaccggtta gcgctagctc attactagcc
     3001 gtccactttc cgctttttct taggatcttc caccttgcgt tttttcttgg gggagccgct
     3061 gcccaggctg gcataatcgg gcacgtcata gggataggcg tcgcctccca gctgagacag
     3121 gtcgatccgt gtctcgtaca ggccggtgat gctctggtgg atcagggtgg cgtccagcac
     3181 ctctttggtg ctggtgtacc tcttccggtc gatggtggtg tcaaagtact tgaaggcggc
     3241 aggggctccc agattggtca gggtaaacag gtggatgata ttctcggcct gctctctgat
     3301 aggcttgtct ctgtgcttgt tgtaggcgct cagcaccttg tccagattag cgtcggccag
     3361 gatcactctc ttggagaact cgctgatctg ctcgatgatc tcgtccaggt agtgtttgtg
     3421 ctgttccaca aacagctgtt tctgctcatt atcctcgggg gagcccttca gcttctcata
     3481 gtggctggcc aggtacagga agttcacata tttggagggc agggccagtt cgtttccctt
     3541 ctgcagttcg ccggcagagg ccagcattct cttccggccg ttttccagct cgaacaggga
     3601 gtacttaggc agcttgatga tcaggtcctt tttcacttct ttgtagccct tggcttccag
     3661 aaagtcgatg ggattcttct cgaagctgct tctttccatg atggtgatcc ccagcagctc
     3721 tttcacactc ttcagtttct tggacttgcc cttttccact ttggccacca ccagcacaga
     3781 ataggccacg gtggggctgt cgaagccgcc gtacttctta gggtcccagt ccttctttct
     3841 ggcgatcagc ttgtcgctgt tcctcttggg caggatagac tctttgctga agccgcctgt
     3901 ctgcacctcg gtctttttca cgatattcac ttggggcata gacagcactt tccgcacggt
     3961 ggcaaagtcc cggcccttat cccacacgat ctcgcctgtt tcgccgtttg tctcgatcag
     4021 aggccgcttc cggatctcgc cgttggccag ggtaatctcg gtcttgaaaa agttcatgat
     4081 gttgctgtag aagaagtact tggcggtagc cttgccgatt tcctgctcgc tcttggcgat
     4141 catcttccgc acgtcgtaca ccttgtagtc gccgtacacg aactcgcttt ccagcttagg
     4201 gtactttttg atcagggcgg ttcccacgac ggcgttcagg taggcgtcgt gggcgtggtg
     4261 gtagttgttg atctcgcgca ctttgtaaaa ctggaaatcc ttccggaaat cggacaccag
     4321 cttggacttc agggtgatca ctttcacttc ccggatcagt ttgtcgttct cgtcgtactt
     4381 agtgttcatc cgggagtcca ggatctgtgc cacgtgcttt gtgatctgcc gggtttccac
     4441 cagctgtctc ttgatgaagc cggccttatc cagttcgctc aggccgcctc tctcggcctt
     4501 ggtcagattg tcgaacttcc tctgggtaat cagcttggca ttcagcagct ggcgccagta
     4561 gttcttcatc ttcttcacga cctcttcgga gggcacgttg tcgctcttgc cccggttctt
     4621 gtcgctccga gtcagcactt tgttatcgat ggagtcgtcc ttcagaaagc tctgaggcac
     4681 gatagcgtcc acatcgtagt cggacagccg gttgatgtcc agttcctggt ccacgtacat
     4741 atcccgccca ttctgcaggt agtacaggta cagcttctcg ttctgcagct gggtgttttc
     4801 cacggggtgt tctttcagga tctggctgcc cagctctttg atgccctctt cgatccgctt
     4861 cattctctcg cggctgttct tctgtccctt ctgggtggtc tggttctctc tggccatttc
     4921 gatcacgatg ttctcgggct tgtgccggcc catcactttc acgagctcgt ccaccacctt
     4981 cactgtctgc aggatgccct tcttaatggc ggggctgccg gccagattgg caatgtgctc
     5041 gtgcaggcta tcgccctggc cggacacctg ggctttctgg atgtcctctt taaaggtcag
     5101 gctgtcgtcg tggatcagct gcatgaagtt tctgttggcg aagccgtcgg acttcaggaa
     5161 atccaggatt gtcttgccgg actgcttgtc ccggatgccg ttgatcagct tccggctcag
     5221 cctgccccag ccggtgtatc tccgccgctt cagctgcttc atcactttgt cgtcgaacag
     5281 gtgggcatag gttttcagcc gttcctcgat catctctctg tcctcaaaca gtgtcagggt
     5341 cagcacgata tcttccagaa tgtcctcgtt ttcctcattg tccaggaagt ccttgtcctt
     5401 gataattttc agcagatcgt ggtatgtgcc cagggaggcg ttgaaccgat cttccacgcc
     5461 ggagatttcc acggagtcga agcactcgat tttcttgaag tagtcctctt tcagctgctt
     5521 cacggtcact ttccggttgg tcttgaacag caggtccacg atggcttttt tctgctcgcc
     5581 gctcaggaag gcgggctttc tcattccctc ggtcacgtat ttcactttgg tcagctcgtt
     5641 gtacacggtg aagtactcgt acagcaggct gtgcttgggc agcaccttct cgttgggcag
     5701 gttcttatcg aagttggtca tccgctcgat gaagctctgg gcgctggcgc ccttgtccac
     5761 cacttcctcg aagttccagg gggtgatggt ttcctcgctc tttctggtca tccaggcgaa
     5821 tctgctgttt cccctggcca gagggcccac gtagtagggg atgcggaagg tcaggatctt
     5881 ctcgatcttt tcccggttgt ccttcaggaa tgggtaaaaa tcttcctgcc gccgcagaat
     5941 ggcgtgcagc tctcccaggt ggatctggtg ggggatgctg ccgttgtcga aggtccgctg
     6001 cttccgcagc aggtcctctc tgttcagctt cacgagcagt tcctcggtgc cgtccatctt
     6061 ttccaggatg ggcttgatga acttgtagaa ctcttcctgg ctggctccgc catcgatgta
     6121 gccggcgtag ccgttcttgc tctggtcgaa gaaaatctct ttgtacttct caggcagctg
     6181 ctgccgcacg agagctttca gcagggtcag gtcctggtgg tgctcgtcgt atctcttgat
     6241 catagaggcg ctcagggggg ccttggtgat ctcggtgttc actctcagga tgtcgctcag
     6301 caggatggcg tcggacaggt tcttggcggc cagaaacagg tcggcgtact ggtcgccgat
     6361 ctgggccagc aggttgtcca ggtcgtcgtc gtaggtgtcc ttgctcagct gcagtttggc
     6421 atcctcggcc aggtcgaagt tgctcttgaa gttgggggtc aggcccaggc tcagggcaat
     6481 caggttgccg aacaggccat tcttcttctc gccgggcagc tgggcgatca gattttccag
     6541 ccgtctgctc ttgctcagtc tggcagacag gatggccttg gcgtccacgc cgctggcgtt
     6601 gatggggttt tcctcgaaca gctggttgta ggtctgcacc agctggatga acagcttgtc
     6661 cacgtcgctg ttgtcggggt tcaggtcgcc ctcgatcagg aagtggcccc ggaacttgat
     6721 catgtgggcc agggccagat agatcagccg caggtcggcc ttgtcggtgc tgtccaccag
     6781 tttctttctc aggtggtaga tggtggggta cttctcgtgg taggccacct cgtccacgat
     6841 gttgccgaag atggggtgcc gctcgtgctt cttatcctct tccaccagga aggactcttc
     6901 cagtctgtgg aagaagctgt cgtccacctt ggccatctcg ttgctgaaga tctcttgcag
     6961 atagcagatc cggttcttcc gtctggtgta tcttcttctg gcggttctct tcagccgggt
     7021 ggcctcggct gtttctccgc tgtcgaacag cagggcgccg atcaggttct tcttgatgct
     7081 gtgccggtcg gtgttgccca gcaccttgaa tttcttgctg ggcaccttgt actcgtcggt
     7141 gatcacggcc cagcccacag agttggtgcc gatggccagg ccgatgctgt acttcttgtc
     7201 catggtggcg atcccgggcc cgcggtaccg caagcttgat atcctgcaga cgcgtcccac
     7261 cttgcgcttc ttcttgggag aacctccaga tccaccaacc actttgtaca agaaagttgg
     7321 gtagagactc tctttcacat cctttggctt ccaaatctgc cctccaatgt ccccattttt
     7381 ttctgcacgg ccacttccca gcacttcctc ttcctccaac catggctcct ttccttgttc
     7441 caacctcaag atcacatcgg gtttggtggt ttccccttgt cccacagaga caaggttgct
     7501 gtaattctcc agcatcacat ccctgtacaa gtttctctgt tcgggattca gccgctgcca
     7561 ctccccctgg gtgaagttca cagtgacatc ctcgaaggtc actcttccct gggaattgtt
     7621 catgccaact tttttgtaca aacttgttga tcccgacagt tagccagaga gctctgctta
     7681 tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac
     7741 gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg
     7801 ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc
     7861 aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     7921 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     7981 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     8041 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     8101 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     8161 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     8221 attactatta ataaccccgg gccccctcga gccggcgcca aagtggatct ctgctgtccc
     8281 tgtaataaac ccgaaaattt tgaatttttg taatttgttt ttgtaattct ttagtttgta
     8341 tgtctgttgc tattatgtct actattcttt cccctgcact gtacccccca atcccccctt
     8401 ttcttttaaa attgtggatg aatactgcca tttgtctcaa gatctagaat tcttactact
     8461 gcagctgcag gccctcccac acataaccag agggcagcaa ttcacgaatc ccaactgccg
     8521 tcggctgtcc atcactgtcc ttcactatgg ctttgatccc aggatgcaga tcgagaagca
     8581 cctgtcggca ccgtccgcag gggctcaaga tgcccctgtt ctcatttccg atcgcgacga
     8641 tacaagtcag gttgccagct gccgcagcag cagcagtgcc cagcaccacg agttctgcac
     8701 aaggtccccc agtaaaatga tatacattga caccagtgaa gatgcggccg tcgctagaga
     8761 gagctgcgct ggcgacgctg tagtcttcag agatggggat gctgttgatt gtagccgttg
     8821 ctctttcaat gagggtggat tcttcttgag acaaaggctt ggccatggtg gtaccccctg
     8881 gggagagagg tcggtgattc ggtcaacgag ggagccgact gccgacgtgc gctccggagg
     8941 cttgcagaat gcggaacacc gcgcgggcag gaacagggcc cacactaccg ccccacaccc
     9001 cgcctcccgc accgcccctt cccggccgct gctctcggcg cgccctgctg agcagccgct
     9061 attggccaca gcccatcgcg gtcggcgcgc tgccattgct ccctggcgct gtccgtctgc
     9121 gagggtacta gtgagacgtg cggcttccgt ttgtcacgtc cggcacgccg cgaaccgcaa
     9181 ggaaccttcc cgacttaggg gcggagcagg aagcgtcgcc ggggggccca caagggtagc
     9241 ggcgaagatc cgggtgacgc tgcgaacgga cgtgaagaat gtgcgagacc cagggtcggc
     9301 gccgctgcgt ttcccggaac cacgcccaga gcagccgcgt ccctgcgcaa acccagggct
     9361 gccttggaaa aggcgcaagg gtcccctcgg ggttgggagg tgggtctgaa acgataatgg
     9421 tgaatatccc tgcctaactc tattcactat agaaagtaca gcaaaaacta ttcttaaacc
     9481 taccaagcct cctactatca ttatgaataa ttttatatac cacagccaat ttgttatgtt
     9541 aaaccaattc cacaaacttg cccatttatc taattccaat aattcttgtt cattcttttc
     9601 ttgctggttt tgcgattctt caattaagga gtgtattaag cttgtgtaat tgttaatttc
     9661 tctgtcccac tccatccagg tcgtgtgatt ccaaatctgt tccagagatt tattactcca
     9721 actagcattc caaggcacag cagtggtgca aatgagtttt ccagagcaac cccaaatccc
     9781 caggagctgt tgatccttta ggtatctttc cacagccagg attcttgcct ggagctgctt
     9841 gatgccccag actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt
     9901 gttctgctgc tgcactatac cagacaataa ttgtctggcc tgtaccgtca gcgtcattga
     9961 cgctgcgccc atagtgcttc ctgctgctcc caagaaccca aggaacaaag ctcctattcc
    10021 cactgctctt ttttctctct gcaccactct tctctttgcc ttggtgggtg ctactcctaa
    10081 tggttcaatt tttactactt tatatttata taattcactt ctccaattgt ccctcatatc
    10141 tcctcctcca ggtctgaaga tcagcggccg cttgctgtgc ggtggtctta cttttgtttt
    10201 gctcttcctc tatcttgtct aaagcttcct tggtgtcttt tatctctatc ctttgatgca
    10261 cacaatagag ggttgctact gtattatata atgatctaag ttcttctgat cctgtctgaa
    10321 gggatggttg tagctgtccc agtatttgtc tacagccttc tgatgtttct aacaggccag
    10381 gattaactgc gaatcgttct agctccctgc ttgcccatac tatatgtttt aatttatatt
    10441 ttttctttcc ccctggcctt aaccgaattt tttcccatcg cgatctaatt ctcccccgct
    10501 taatactgac gctctcgcac ccatctctct ccttctagcc tccgctagtc aaaatttttg
    10561 gcgtactcac cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca agccgagtcc
    10621 tgcgtcgaga gagctcctct ggtttccctt tcgctttcaa gtccctgttc gggcgccact
    10681 gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac cagagtcaca
    10741 caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag gcttaagcag
    10801 tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc
    10861 gtttatgtat cgagctaggc acttaaatac aatatctctg caatgcggca attcagtggt
    10921 tcgtccaatc catgtcagac ccgtctgttg ccttcctaat aaggcacgat cgtaccacct
    10981 tacttccacc aatcggcatg cacggtgctt tttctctcct tgtaaggcat gttgctaact
    11041 catcgttacc atgttgcaag actacaagag tattgcataa gactacatta agcttgcagc
    11101 tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag
    11161 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
    11221 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
    11281 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
    11341 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
    11401 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
    11461 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
    11521 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
    11581 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
    11641 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
    11701 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
    11761 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
    11821 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
    11881 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
    11941 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
    12001 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
    12061 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
    12121 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
    12181 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
    12241 acctagatcc ttttaaatta aaaatgaagt tttaaatca
//
