LOCUS       sequence-447848-       12441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for CRISPRi integration at AAVS1 with a GFP 
            fluorophore.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12441)
  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 12441)
  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-GFP
FEATURES             Location/Qualifiers
     source          1..12441
                     /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..2685
                     /label=chicken beta-actin promoter
     intron          2686..3702
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3626..3648
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3710..3729
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3761..3770
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3767..3793,3794..3796,3797..6283,6284..6286,
                     6287..7870)
                     /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             3767..7870
                     /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             7874..7900
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7880..7906
                     /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             7919..7939
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7946..7966
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8276..8461
                     /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             8480..8536
                     /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             join(8537..8539,8540..9253)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(8840..8859)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9187..9208
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(9342..9361)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    9407..9462
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(9461..9480)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    complement(9811..9844)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    9885..10721
                     /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(10769..11357)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10849..10868)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11528..12319,12320..12388))
                     /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     12151..12170
                     /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 tggggggggg gggggggggg
     2521 gggggggggc gcgcgcgagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg
     2581 agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg
     2641 cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc
     2701 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
     2761 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
     2821 cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc
     2881 cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg
     2941 gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg
     3001 gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg
     3061 tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga
     3121 gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt
     3181 gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc
     3241 gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc
     3301 ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg
     3361 gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg
     3421 tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc
     3481 ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg
     3541 ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc
     3601 cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga
     3661 gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg
     3721 gttattgtgc tgtctcatca ttttggcaaa gaatgtcgac gccaccatgg acaagaagta
     3781 cagcatcggc ctggccatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta
     3841 caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa
     3901 gaacctgatc ggcgccctgc tgttcgacag cggagaaaca gccgaggcca cccggctgaa
     3961 gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat
     4021 cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt
     4081 cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga
     4141 ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag
     4201 caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg
     4261 gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt
     4321 catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg
     4381 cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct
     4441 gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggcaacctga ttgccctgag
     4501 cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca
     4561 gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca
     4621 gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat
     4681 cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata
     4741 cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga
     4801 gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct acatcgatgg
     4861 cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg
     4921 caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt
     4981 cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg
     5041 gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac
     5101 cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat
     5161 gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg
     5221 cgccagcgcc cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga
     5281 gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtaca acgagctgac
     5341 caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa
     5401 aaaagccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa
     5461 agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga
     5521 tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga
     5581 cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact
     5641 gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga
     5701 caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa
     5761 gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc
     5821 cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa
     5881 agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc
     5941 caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga
     6001 cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag
     6061 agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga
     6121 agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca
     6181 gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga
     6241 ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gacgctatcg tgcctcagag
     6301 ctttctgaag gacgactcca tcgataacaa agtgctgact cggagcgaca agaaccgggg
     6361 caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcgcca
     6421 gctgctgaat gccaagctga ttacccagag gaagttcgac aatctgacca aggccgagag
     6481 aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg
     6541 gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga
     6601 gaacgacaaa ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga
     6661 tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca
     6721 cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga
     6781 aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag
     6841 cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt
     6901 tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac
     6961 aaacggcgaa acaggcgaga tcgtgtggga taagggccgg gactttgcca ccgtgcggaa
     7021 agtgctgtct atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt
     7081 cagcaaagag tctatcctgc ccaagaggaa cagcgacaag ctgatcgcca gaaagaagga
     7141 ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt
     7201 ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg
     7261 gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa
     7321 gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga
     7381 gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga
     7441 actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct
     7501 gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc acaaacacta
     7561 cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc
     7621 taatctggac aaggtgctga gcgcctacaa caagcacaga gacaagccta tcagagagca
     7681 ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa
     7741 gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc
     7801 caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct
     7861 gggaggcgac gcctatccct atgacgtgcc cgattatgcc agcctgggca gcggctcccc
     7921 caagaaaaaa cgcaaggtgg aagatcctaa gaaaaagcgg aaagtggacg gcattggtag
     7981 tgggagcaac ggcagcagcg gatccggagg gccgagctct ggcgcacccc caccaagtgg
     8041 agggtctcct gccgggtccc caacatctac tgaagaaggc accagcgaat ccgcaacgcc
     8101 cgagtcaggc cctggtacct ccacagaacc atctgaaggt agtgcgcctg gttccccagc
     8161 tggaagccct acttccaccg aagaaggcac gtcaaccgaa ccaagtgaag gatctgcccc
     8221 tgggaccagc actgaaccat ctgagatgga tgctaagtca ctaactgcct ggtcccggac
     8281 actggtgacc ttcaaggatg tatttgtgga cttcaccagg gaggagtgga agctgctgga
     8341 cactgctcag cagatcgtgt acagaaatgt gatgctggag aactataaga acctggtttc
     8401 cttgggttat cagcttacta agccagatgt gatcctccgg ttggagaagg gagaagagcc
     8461 cagcggccgc ggtagtgggg ctactaactt cagcctgctg aagcaggctg gagacgtgga
     8521 ggagaaccct ggacctgtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     8581 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtctggcg agggcgaggg
     8641 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     8701 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     8761 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     8821 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     8881 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     8941 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     9001 caagcagaag aacggcatca aggcgaactt caagatccgc cacaacatcg aggacggcag
     9061 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     9121 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     9181 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     9241 gctgtacaag taacgcgtga attcactcct caggtgcagg ctgcctatca gaaggtggtg
     9301 gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa
     9361 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt
     9421 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg
     9481 agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc
     9541 catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag
     9601 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
     9661 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
     9721 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta
     9781 tggagatccc tcgacctgca gcccaagctt ataacttcgt atagcataca ttatacgaag
     9841 ttatgcggcc gcaccggtat gcatttaatt aagatatcag ctttactagg gacaggattg
     9901 gtgacagaaa agccccatcc ttaggcctcc tccttcctag tctcctgata ttgggtctaa
     9961 cccccacctc ctgttaggca gattccttat ctggtgacac acccccattt cctggagcca
    10021 tctctctcct tgccagaacc tctaaggttt gcttacgatg gagccagaga ggatcctggg
    10081 agggagagct tggcaggggg tgggagggaa gggggggatg cgtgacctgc ccggttctca
    10141 gtggccaccc tgcgctaccc tctcccagaa cctgagctgc tctgacgcgg ctgtctggtg
    10201 cgtttcactg atcctggtgc tgcagcttcc ttacacttcc caagaggaga agcagtttgg
    10261 aaaaacaaaa tcagaataag ttggtcctga gttctaactt tggctcttca cctttctagt
    10321 ccccaattta tattgttcct ccgtgcgtca gttttacctg tgagataagg ccagtagcca
    10381 gccccgtcct ggcagggctg tggtgaggag gggggtgtcc gtgtggaaaa ctccctttgt
    10441 gagaatggtg cgtcctaggt gttcaccagg tcgtggccgc ctctactccc tttctctttc
    10501 tccatccttc tttccttaaa gagtccccag tgctatctgg gacatattcc tccgcccaga
    10561 gcagggtccc gcttccctaa ggccctgctc tgggcttctg ggtttgagtc cttggcaagc
    10621 ccaggagagg cgctcaggct tccctgtccc ccttcctcgt ccaccatctc atgcccctgg
    10681 ctctcctgcc ccttccctac aggggttcct ggctctgctc taagggcgaa ttcgccagga
    10741 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
    10801 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
    10861 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
    10921 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
    10981 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
    11041 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
    11101 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
    11161 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
    11221 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
    11281 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
    11341 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
    11401 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
    11461 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
    11521 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
    11581 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
    11641 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
    11701 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
    11761 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
    11821 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
    11881 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
    11941 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
    12001 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
    12061 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
    12121 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
    12181 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
    12241 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
    12301 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
    12361 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
    12421 atcagggttg aattcgccct t
//
