LOCUS       sequence-448471-       12440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for CRISPRi integration at AAVS1 with an mCherry 
            fluorophore.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12440)
  AUTHORS   She R, Fair T, Schaefer NK, Saunders RA, Pavlovic BJ, Weissman JS, 
            Pollen AA
  TITLE     Comparative landscape of genetic dependencies in human and 
            chimpanzee stem cells.
  JOURNAL   Cell. 2023 Jul 6;186(14):2977-2994.e23. doi: 
            10.1016/j.cell.2023.05.043. Epub 2023 Jun 20.
  PUBMED    37343560
REFERENCE   2  (bases 1 to 12440)
  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.2023.05.043"; journalName: "Cell"; date: "2023-07-6-
            6"; volume: "186"; issue: "14"; pages: "2977-2994.e23"
COMMENT     Sequence Label: AAVS1_dCas9-XTEN-KRAB-p2A-mCherry
FEATURES             Location/Qualifiers
     source          1..12440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..804
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     protein_bind    complement(806..839)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    846..871
                     /label=SA
                     /note="splice acceptor site"
     CDS             895..948
                     /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"
     regulatory      964..973
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             970..1773
                     /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(1033..1052)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1643..1662
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1793..1810)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1799..2023
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        2029..2408
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2410..2687
                     /label=chicken beta-actin promoter
     intron          2688..3704
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3628..3650
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3712..3731
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3763..3772
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3769..3795,3796..3798,3799..6285,6286..6288,
                     6289..7872)
                     /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             3769..7872
                     /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             7876..7902
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7882..7908
                     /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             7921..7941
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7948..7968
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8278..8463
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             8482..8538
                     /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             8539..9249
                     /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(8684..8702)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     8952..8971
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(9341..9360)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    9406..9461
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(9460..9479)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    complement(9810..9843)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    9884..10720
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     rep_origin      complement(10768..11356)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10848..10867)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11527..12318,12319..12387))
                     /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     12150..12169
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tgctttctct gaccagcatt ctctcccctg ggcctgtgcc gctttctgtc tgcagcttgt
       61 ggcctgggtc acctctacgg ctggcccaga tccttccctg ccgcctcctt caggttccgt
      121 cttcctccac tccctcttcc ccttgctctc tgctgtgttg ctgcccaagg atgctctttc
      181 cggagcactt ccttctcggc gctgcaccac gtgatgtcct ctgagcggat cctccccgtg
      241 tctgggtcct ctccgggcat ctctcctccc tcacccaacc ccatgccgtc ttcactcgct
      301 gggttccctt ttccttctcc ttctggggcc tgtgccatct ctcgtttctt aggatggcct
      361 tctccgacgg atgtctccct tgcgtcccgc ctccccttct tgtaggcctg catcatcacc
      421 gtttttctgg acaaccccaa agtaccccgt ctccctggct ttagccacct ctccatcctc
      481 ttgctttctt tgcctggaca ccccgttctc ctgtggattc gggtcacctc tcactccttt
      541 catttgggca gctcccctac cccccttacc tctctagtct gtgctagctc ttccagcccc
      601 ctgtcatggc atcttccagg ggtccgagag ctcagctagt cttcttcctc caacccgggc
      661 ccctatgtcc acttcaggac agcatgtttg ctgcctccag ggatcctgtg tccccgagct
      721 gggaccacct tatattccca gggccggtta atgtggctct ggttctgggt acttttatct
      781 gtcccctcca ccccacagtg gggcaataac ttcgtatagc atacattata cgaagttata
      841 agcttctgac ctcttctctt cctcccacag ggcctcgaga gatctggcag cggagagggc
      901 agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctag gctcgagggt
      961 accgccacca tgggatcggc cattgaacaa gatggattgc acgcaggttc tccggccgct
     1021 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     1081 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     1141 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     1201 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     1261 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     1321 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     1381 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     1441 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     1501 aaggcgcgca tgcccgacgg cgatgatctc gtcgtgaccc atggcgatgc ctgcttgccg
     1561 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     1621 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     1681 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     1741 gccttctatc gccttcttga cgagttcttc tgagtttaaa cccgctgatc agcctcgact
     1801 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     1861 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     1921 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     1981 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tgggtctcga cattgattat
     2041 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     2101 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     2161 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     2221 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     2281 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     2341 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     2401 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
     2461 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg
     2521 gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc
     2581 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
     2641 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc
     2701 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
     2761 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
     2821 agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc
     2881 tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg
     2941 tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg
     3001 cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc
     3061 ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt
     3121 gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag
     3181 ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg
     3241 ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg
     3301 ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg
     3361 cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt
     3421 tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc
     3481 gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt
     3541 cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct
     3601 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
     3661 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
     3721 tggttattgt gctgtctcat cattttggca aagaatgtcg acgccaccat ggacaagaag
     3781 tacagcatcg gcctggccat cggcaccaac tctgtgggct gggccgtgat caccgacgag
     3841 tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag
     3901 aagaacctga tcggcgccct gctgttcgac agcggagaaa cagccgaggc cacccggctg
     3961 aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag
     4021 atcttcagca acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc
     4081 ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac
     4141 gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac
     4201 agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc
     4261 cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg
     4321 ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc
     4381 ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat
     4441 ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggcaacct gattgccctg
     4501 agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg
     4561 cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac
     4621 cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac
     4681 atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga
     4741 tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct
     4801 gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacatcgat
     4861 ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac
     4921 ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc
     4981 ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg
     5041 cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg
     5101 accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg
     5161 atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag
     5221 ggcgccagcg cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac
     5281 gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta caacgagctg
     5341 accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag
     5401 aaaaaagcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg
     5461 aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa
     5521 gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag
     5581 gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca
     5641 ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac
     5701 gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg
     5761 aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag
     5821 tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt
     5881 aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt
     5941 gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg
     6001 gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc
     6061 agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc
     6121 gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc
     6181 cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg
     6241 gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggacgctat cgtgcctcag
     6301 agctttctga aggacgactc catcgataac aaagtgctga ctcggagcga caagaaccgg
     6361 ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgc
     6421 cagctgctga atgccaagct gattacccag aggaagttcg acaatctgac caaggccgag
     6481 agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc
     6541 cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac
     6601 gagaacgaca aactgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc
     6661 gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc
     6721 cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg
     6781 gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag
     6841 agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac
     6901 tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag
     6961 acaaacggcg aaacaggcga gatcgtgtgg gataagggcc gggactttgc caccgtgcgg
     7021 aaagtgctgt ctatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc
     7081 ttcagcaaag agtctatcct gcccaagagg aacagcgaca agctgatcgc cagaaagaag
     7141 gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg
     7201 gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg
     7261 gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc
     7321 aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc
     7381 gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac
     7441 gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag
     7501 ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaaacac
     7561 tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac
     7621 gctaatctgg acaaggtgct gagcgcctac aacaagcaca gagacaagcc tatcagagag
     7681 caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc
     7741 aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac
     7801 gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag
     7861 ctgggaggcg acgcctatcc ctatgacgtg cccgattatg ccagcctggg cagcggctcc
     7921 cccaagaaaa aacgcaaggt ggaagatcct aagaaaaagc ggaaagtgga cggcattggt
     7981 agtgggagca acggcagcag cggatccgga gggccgagct ctggcgcacc cccaccaagt
     8041 ggagggtctc ctgccgggtc cccaacatct actgaagaag gcaccagcga atccgcaacg
     8101 cccgagtcag gccctggtac ctccacagaa ccatctgaag gtagtgcgcc tggttcccca
     8161 gctggaagcc ctacttccac cgaagaaggc acgtcaaccg aaccaagtga aggatctgcc
     8221 cctgggacca gcactgaacc atctgagatg gatgctaagt cactaactgc ctggtcccgg
     8281 acactggtga ccttcaagga tgtatttgtg gacttcacca gggaggagtg gaagctgctg
     8341 gacactgctc agcagatcgt gtacagaaat gtgatgctgg agaactataa gaacctggtt
     8401 tccttgggtt atcagcttac taagccagat gtgatcctcc ggttggagaa gggagaagag
     8461 cccagcggcc gcggtagtgg ggctactaac ttcagcctgc tgaagcaggc tggagacgtg
     8521 gaggagaacc ctggacctat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
     8581 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
     8641 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     8701 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
     8761 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
     8821 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
     8881 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     8941 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
     9001 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
     9061 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     9121 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     9181 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     9241 tacaagtaat aacgcgtgaa ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg
     9301 ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa
     9361 attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta
     9421 ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga
     9481 gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc
     9541 atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc
     9601 cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta
     9661 tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac
     9721 tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat
     9781 ggagatccct cgacctgcag cccaagctta taacttcgta tagcatacat tatacgaagt
     9841 tatgcggccg caccggtatg catttaatta agatatcagc tttactaggg acaggattgg
     9901 tgacagaaaa gccccatcct taggcctcct ccttcctagt ctcctgatat tgggtctaac
     9961 ccccacctcc tgttaggcag attccttatc tggtgacaca cccccatttc ctggagccat
    10021 ctctctcctt gccagaacct ctaaggtttg cttacgatgg agccagagag gatcctggga
    10081 gggagagctt ggcagggggt gggagggaag ggggggatgc gtgacctgcc cggttctcag
    10141 tggccaccct gcgctaccct ctcccagaac ctgagctgct ctgacgcggc tgtctggtgc
    10201 gtttcactga tcctggtgct gcagcttcct tacacttccc aagaggagaa gcagtttgga
    10261 aaaacaaaat cagaataagt tggtcctgag ttctaacttt ggctcttcac ctttctagtc
    10321 cccaatttat attgttcctc cgtgcgtcag ttttacctgt gagataaggc cagtagccag
    10381 ccccgtcctg gcagggctgt ggtgaggagg ggggtgtccg tgtggaaaac tccctttgtg
    10441 agaatggtgc gtcctaggtg ttcaccaggt cgtggccgcc tctactccct ttctctttct
    10501 ccatccttct ttccttaaag agtccccagt gctatctggg acatattcct ccgcccagag
    10561 cagggtcccg cttccctaag gccctgctct gggcttctgg gtttgagtcc ttggcaagcc
    10621 caggagaggc gctcaggctt ccctgtcccc cttcctcgtc caccatctca tgcccctggc
    10681 tctcctgccc cttccctaca ggggttcctg gctctgctct aagggcgaat tcgccaggaa
    10741 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
    10801 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
    10861 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
    10921 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
    10981 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
    11041 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
    11101 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
    11161 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
    11221 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
    11281 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
    11341 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
    11401 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
    11461 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
    11521 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
    11581 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
    11641 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
    11701 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
    11761 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
    11821 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
    11881 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
    11941 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
    12001 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
    12061 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
    12121 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
    12181 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
    12241 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
    12301 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
    12361 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
    12421 tcagggttga attcgccctt
//
