LOCUS       sequence-434911-       14055 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline-inducible (Zim3)KRAB-dCas9-HA-P2A-mCherry cassette for 
            integration at the AAVS1-locus of the human genome..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14055)
  AUTHORS   Edenhofer FC, Termeg A, Ohnuki M, Jocher J, Kliesmete Z, Briem E, 
            Hellmann I, Enard W
  TITLE     Generation and characterization of inducible KRAB-dCas9 iPSCs from 
            primates for cross-species CRISPRi.
  JOURNAL   iScience. 2024 May 23;27(6):110090. doi: 10.1016/j.isci.2024.110090.
            eCollection 2024 Jun 21.
  PUBMED    38947524
REFERENCE   2  (bases 1 to 14055)
  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: "iScience.";
            date: "2024-05-23"; volume: "27(6):110090. doi"; pages: " 
            10.1016/j.isci.2024.110090. eCollection 2024 Jun 21"
COMMENT     Sequence Label: pAAVS1-TetOn-ZIM3-KRAB-dCas9-P2A-mCherry
FEATURES             Location/Qualifiers
     source          1..14055
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      46..55
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(52..78,79..81,82..2568,2569..2571,2572..4155)
                     /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             52..4155
                     /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             4159..4185
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4165..4191
                     /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             4204..4224
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4231..4251
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4270..4326
                     /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             4327..5037
                     /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(4472..4490)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4740..4759
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5069..5190
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5106..5125)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5160..5179
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5239..5255)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5239..5255)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5252..5274)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5263..5279
                     /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(5287..5293,5294..5311,5312..5317))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5332..5353
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5470..5487)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5543..5576
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     5907..5926
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    5925..5980
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     6026..6045
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(6132..6878)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(6910..6929)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          6937..7953
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(6991..7013)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     enhancer        8233..8612
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    8618..8842
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     8831..8848
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(8868..9671)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8979..8998)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     9589..9608
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     regulatory      9668..9677
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(9693..9746)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    9770..9795
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    9802..9835
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    9837..10640
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     CDS             join(10694..10762,10763..11554)
                     /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(10912..10931)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11725..12313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12214..12233
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    12361..13197
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     protein_bind    13230..13248
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    13266..13284
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    13302..13320
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    13338..13356
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    13374..13392
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    13410..13428
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    13446..13464
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     13509..13533
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    13615..13639
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(13988..14012)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             14032..14052
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 cgcgtctgca ggatatcaag cttgcggtac cgcgggcccg ggatcgccac catggacaag
       61 aagtacagca tcggcctggc catcggcacc aactctgtgg gctgggccgt gatcaccgac
      121 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
      181 aagaagaacc tgatcggcgc cctgctgttc gacagcggag aaacagccga ggccacccgg
      241 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
      301 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
      361 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
      421 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
      481 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
      541 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
      601 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
      661 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa
      721 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggcaa cctgattgcc
      781 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
      841 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
      901 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
      961 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
     1021 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
     1081 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatc
     1141 gatggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
     1201 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg
     1261 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
     1321 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
     1381 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
     1441 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
     1501 aagggcgcca gcgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
     1561 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtacaacgag
     1621 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
     1681 cagaaaaaag ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
     1741 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
     1801 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
     1861 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
     1921 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
     1981 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
     2041 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
     2101 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
     2161 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
     2221 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
     2281 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
     2341 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
     2401 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
     2461 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
     2521 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacgc tatcgtgcct
     2581 cagagctttc tgaaggacga ctccatcgat aacaaagtgc tgactcggag cgacaagaac
     2641 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
     2701 cgccagctgc tgaatgccaa gctgattacc cagaggaagt tcgacaatct gaccaaggcc
     2761 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
     2821 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
     2881 gacgagaacg acaaactgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
     2941 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
     3001 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
     3061 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
     3121 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
     3181 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
     3241 gagacaaacg gcgaaacagg cgagatcgtg tgggataagg gccgggactt tgccaccgtg
     3301 cggaaagtgc tgtctatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
     3361 ggcttcagca aagagtctat cctgcccaag aggaacagcg acaagctgat cgccagaaag
     3421 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
     3481 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
     3541 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
     3601 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
     3661 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga
     3721 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
     3781 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaaa
     3841 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
     3901 gacgctaatc tggacaaggt gctgagcgcc tacaacaagc acagagacaa gcctatcaga
     3961 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
     4021 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
     4081 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct
     4141 cagctgggag gcgacgccta tccctatgac gtgcccgatt atgccagcct gggcagcggc
     4201 tcccccaaga aaaaacgcaa ggtggaagat cctaagaaaa agcggaaagt ggacggcatt
     4261 ggtagtgggg ctactaactt cagcctgctg aagcaggctg gagacgtgga ggagaaccct
     4321 ggacctatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     4381 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     4441 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     4501 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     4561 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     4621 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     4681 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     4741 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     4801 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     4861 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     4921 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     4981 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaaacc
     5041 ggtgcggccg caatcgatcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5101 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5161 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5221 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5281 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5341 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5401 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5461 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5521 agctcactca aaggacggcc gcataacttc gtataatgta tgctatacga agttataagc
     5581 ttgggctgca ggtcgaggga tctccataag agaagaggga cagctatgac tgggagtagt
     5641 caggagagga ggaaaaatct ggctagtaaa acatgtaagg aaaattttag ggatgttaaa
     5701 gaaaaaaata acacaaaaca aaatataaaa aaaatctaac ctcaagtcaa ggcttttcta
     5761 tggaataagg aatggacagc agggggctgt ttcatatact gatgacctct ttatagccaa
     5821 cctttgttca tggcagccag catatgggca tatgttgcca aactctaaac caaatactca
     5881 ttctgatgtt ttaaatgatt tgccctccca tatgtccttc cgagtgagag acacaaaaaa
     5941 ttccaacaca ctattgcaat gaaaataaat ttcctttatt agccagaagt cagatgctca
     6001 aggggcttca tgatgtcccc ataatttttg gcagagggaa aaagatctca gtggtatttg
     6061 tgagccaggg cattggccac accagccacc accttctgat aggcagcctg cacctgagga
     6121 gtgaattcat tttagcctgg cagcatatcg agatcgaagt catcgagagc gtcagcaggg
     6181 agcatatcca gatcaaaatc atccagggca tcggcgggca gcatgtccag gtcgaagtcg
     6241 tccagggcgt cggtgggtcc gccgctctcg cacttcagct gcttctcgag gccgcagatg
     6301 atcagttcca ggccgaacag gaaggcgggc tcggcgccct gtctgtcgaa cagctcgatg
     6361 gcctgcttca gcaggggggg catgctgtcg gtggtgggtg tctctctttc ctctttggcg
     6421 acctggtgtt cctgttcttc cagcacgcag cccagggtga agtggcccac ggcgctcagg
     6481 gcgtacaggg cgttttccag gctgaagccc tgctggcaca ggaaggccag ctggttttcc
     6541 agtgtctcgt actgcttctc ggtgggtctg gtgcccaggt gcactttggc gccgtcccgg
     6601 tggctcagca gggcgcatct gtagctcttg gcgttgttcc gcaggaagtc ctgccagctc
     6661 tcgccttcca gagggcagct gtgggtgtgg tgccggtcca gcatctcgat gggcagggcg
     6721 tccagcaggg cccgcttgtt cttcacgtgc cagtacaggg tgggctgttc cacgcccagt
     6781 ttctgggcca gcttccgggt ggtcaggccc tcgatgccca cgccgttcag cagttccagg
     6841 gcgctgttga tcactttgct cttgtccagc cggctcatgt cgacattctt tgccaaaatg
     6901 atgagacagc acaataacca gcacgttgcc caggagctgt aggaaaaaga agaaggcatg
     6961 aacatggtta gcagaggctc tagagccgcc ggtcacacgc cagaagccga accccgccct
     7021 gccccgtccc ccccgaaggc agccgtcccc ccgcggacag ccccgaggct ggagagggag
     7081 aaggggacgg cggcgcggcg acgcacgaag gccctccccg cccatttcct tcctgccggc
     7141 gccgcaccgc ttcgccccgc gcccgctaga gggggtgcgg cggcgcctcc cagatttcgg
     7201 ctccgcacag atttgggaca aaggaagtcc ctgcgccctc tcgcacgatt accataaaag
     7261 gcaatggctg cggctcgccg cgcctcgaca gccgccggcg ctccgggggc cgccgcgccc
     7321 cycccccgag cccyccccgg cccgmggcgg ccccgccccg cccggcaccc ccmcctgccg
     7381 ccaccccccg cccggcacgg cgagccccgc gccacgcccc gtacggagcc ccgcacccga
     7441 agccgggccg tgctcagcaa ctcggggagg ggggtgcagg gggggttgca gcccgaccga
     7501 cgcgcccaca ccccctgctc acccccccac gcacacaccc cgcacgcagc ctttgttccc
     7561 ctcgcagccc cccccgcacc gcggggcacc gcccccggcc gcgctcccct cgcgcacact
     7621 gcggagcgca caaagccccg cgccgcgccc gcagcgctca cagccgccgg gcagcgcgga
     7681 gccgcacgcg gcgctcccca cgcacacaca cacgcacgca ccccccgagc cgctcccccc
     7741 gcacaaaggg ccctcccgga gcccctcaag gctttcacgc agccacagaa aagaaacaag
     7801 ccgtcattaa accaagcgct aattacagcc cggaggagaa gggccgtccc gcccgctcac
     7861 ctgtgggagt aacgcggtca gtcagagccg gggcgggcgg cgcgaggcgg cggcggagcg
     7921 gggcacgggg cgaaggcagc gcgcagcgac tcccgcccgc cgcgcgcttc gctttttata
     7981 gggccgccgc cgccgccgcc tcgccataaa aggaaacttt cggagcgcgc cgctctgatt
     8041 ggctgccgcc gcmccycycc sccycscccc cccccccccc cccccccccc cccccccccc
     8101 yggcgcgcsc cccccccccc cccccgcccc catcgctgca caaaataatt aaaaaataaa
     8161 taaatacaaa attgggggtg gggagggggg ggagatgggg agagtgaagc agaacgtggg
     8221 gctcacctcg accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
     8281 gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
     8341 aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
     8401 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
     8461 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
     8521 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
     8581 ttactattaa taactagtca ataatcaatg tcgagaccca tagagcccac cgcatcccca
     8641 gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc
     8701 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
     8761 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
     8821 ggctggcaac tagaaggcac agtcgaggct gatcagcggg tttaaactca gaagaactcg
     8881 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     8941 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
     9001 atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     9061 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcatcg
     9121 ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
     9181 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     9241 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     9301 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     9361 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     9421 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc
     9481 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     9541 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     9601 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatg
     9661 gccgatccca tggtggcggt accctcgagc ctagggccgg gattctcctc cacgtcaccg
     9721 catgttagaa gacttcctct gccctctccg ctgccagatc tctcgaggcc ctgtgggagg
     9781 aagagaagag gtcagaagct tataacttcg tataatgtat gctatacgaa gttattgccc
     9841 cactgtgggg tggaggggac agataaaagt acccagaacc agagccacat taaccggccc
     9901 tgggaatata aggtggtccc agctcgggga cacaggatcc ctggaggcag caaacatgct
     9961 gtcctgaagt ggacataggg gcccgggttg gaggaagaag actagctgag ctctcggacc
    10021 cctggaagat gccatgacag ggggctggaa gagctagcac agactagaga ggtaaggggg
    10081 gtaggggagc tgcccaaatg aaaggagtga gaggtgaccc gaatccacag gagaacgggg
    10141 tgtccaggca aagaaagcaa gaggatggag aggtggctaa agccagggag acggggtact
    10201 ttggggttgt ccagaaaaac ggtgatgatg caggcctaca agaaggggag gcgggacgca
    10261 agggagacat ccgtcggaga aggccatcct aagaaacgag agatggcaca ggccccagaa
    10321 ggagaaggaa aagggaaccc agcgagtgaa gacggcatgg ggttgggtga gggaggagag
    10381 atgcccggag aggacccaga cacggggagg atccgctcag aggacatcac gtggtgcagc
    10441 gccgagaagg aagtgctccg gaaagagcat ccttgggcag caacacagca gagagcaagg
    10501 ggaagaggga gtggaggaag acggaacctg aaggaggcgg cagggaagga tctgggccag
    10561 ccgtagaggt gacccaggcc acaagctgca gacagaaagc ggcacaggcc caggggagag
    10621 aatgctggtc agagaaagca aagggcgaat tcaaccctga taaatgcttc aataatattg
    10681 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
    10741 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
    10801 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
    10861 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
    10921 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
    10981 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
    11041 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
    11101 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
    11161 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
    11221 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
    11281 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
    11341 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
    11401 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
    11461 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
    11521 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
    11581 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
    11641 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
    11701 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
    11761 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
    11821 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
    11881 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
    11941 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
    12001 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
    12061 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
    12121 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
    12181 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
    12241 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
    12301 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcga attcgccctt
    12361 agagcagagc caggaacccc tgtagggaag gggcaggaga gccaggggca tgagatggtg
    12421 gacgaggaag ggggacaggg aagcctgagc gcctctcctg ggcttgccaa ggactcaaac
    12481 ccagaagccc agagcagggc cttagggaag cgggaccctg ctctgggcgg aggaatatgt
    12541 cccagatagc actggggact ctttaaggaa agaaggatgg agaaagagaa agggagtaga
    12601 ggcggccacg acctggtgaa cacctaggac gcaccattct cacaaaggga gttttccaca
    12661 cggacacccc cctcctcacc acagccctgc caggacgggg ctggctactg gccttatctc
    12721 acaggtaaaa ctgacgcacg gaggaacaat ataaattggg gactagaaag gtgaagagcc
    12781 aaagttagaa ctcaggacca acttattctg attttgtttt tccaaactgc ttctcctctt
    12841 gggaagtgta aggaagctgc agcaccagga tcagtgaaac gcaccagaca gccgcgtcag
    12901 agcagctcag gttctgggag agggtagcgc agggtggcca ctgagaaccg ggcaggtcac
    12961 gcatcccccc cttccctccc accccctgcc aagctctccc tcccaggatc ctctctggct
    13021 ccatcgtaag caaaccttag aggttctggc aaggagagag atggctccag gaaatggggg
    13081 tgtgtcacca gataaggaat ctgcctaaca ggaggtgggg gttagaccca atatcaggag
    13141 actaggaagg aggaggccta aggatggggc ttttctgtca ccaatcctgt ccctagtaaa
    13201 gctgatatct ccaccccaca gtggtttact ccctatcagt gatagagaac gtatgaagag
    13261 tttactccct atcagtgata gagaacgtat gcagacttta ctccctatca gtgatagaga
    13321 acgtataagg agtttactcc ctatcagtga tagagaacgt atgaccagtt tactccctat
    13381 cagtgataga gaacgtatct acagtttact ccctatcagt gatagagaac gtatatccag
    13441 tttactccct atcagtgata gagaacgtat aagctttagg cgtgtacggt gggcgcctat
    13501 aaaagcagag ctcgtttagt gaaccgtcag atcgcctgga gcaattccac aacacttttg
    13561 tcttatacca actttccgta ccacttccta ccctcgtaaa cttaaggtta atcaacaagt
    13621 ttgtacaaaa aagcaggctg gcgccggaac caattcagtc gacaggccta ggctacgcgc
    13681 gccaccatga acaattccca gggaagagtg accttcgagg atgtcactgt gaacttcacc
    13741 cagggggagt ggcagcggct gaatcccgaa cagagaaact tgtacaggga tgtgatgctg
    13801 gagaattaca gcaaccttgt ctctgtggga caaggggaaa ccaccaaacc cgatgtgatc
    13861 ttgaggttgg aacaaggaaa ggagccatgg ttggaggaag aggaagtgct gggaagtggc
    13921 cgtgcagaaa aaaatgggga cattggaggg cagatttgga agccaaagga tgtgaaagag
    13981 agtctctacc caactttctt gtacaaagtg gttggtggat ctggaggttc tcccaagaag
    14041 aagcgcaagg tggga
//
