LOCUS       sequence-194744-       15533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EBNA episome plasmid for CAG promoter constitutive expression of 
            dCas9-GFP. Includes p53 shRNA expression cassette..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15533)
  AUTHORS   Weltner J, Balboa D, Katayama S, Bespalov M, Krjutskov K, Jouhilahti
            EM, Trokovic R, Kere J, Otonkoski T
  TITLE     Human pluripotent reprogramming with CRISPR activators.
  JOURNAL   Nat Commun. 2018 Jul 6;9(1):2643. doi: 10.1038/s41467-018-05067-x.
  PUBMED    29980666
REFERENCE   2  (bases 1 to 15533)
  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: "Nat 
            Commun."; date: "2018-07-6"; volume: "9(1):2643. doi"; pages: " 
            10.1038/s41467-018-05067-x"
COMMENT     Sequence Label: pCXLE-dCas9GFP-shP53
FEATURES             Location/Qualifiers
     source          1..15533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1752..1778
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1782..1802
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1812..1835,1836..1838,1839..4394,4395..4397,
                     4398..5912)
                     /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 N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(1812..1835,1836..1838,1839..4325,4326..4328,
                     4329..5912)
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5916..5936
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(5961..5963,5964..5966,5967..6680)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5961..5963,5964..5966,5967..6677)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6267..6286)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6614..6635
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6718..7306
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6771..6791)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7189..7200)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7396..7415)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7461..7516
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7515..7534)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7877..7893)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7877..7893)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7890..7912)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7901..7917
                     /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(7925..7931,7932..7949,7950..7955))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7970..7991
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    8080..8113
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             8574..10499
                     /codon_start=1
                     /product="Epstein-Barr nuclear antigen 1, also known as 
                     EBNA-1"
                     /label=EBNA1
                     /note="crucial for latent viral infection, and for episomal
                     amplification of vectors containing the oriP origin (Okita 
                     et al., 2013)"
                     /translation="MSDEGPGTGPGNGLGEKGDTSGPEGSGGSGPQRRGGDNHGRGRGR
                     GRGRGGGRPGAPGGSGSGPRHRDGVRRPQKRPSCIGCKGTHGGTGAGAGAGGAGAGGAG
                     AGGGAGAGGGAGGAGGAGGAGAGGGAGAGGGAGGAGGAGAGGGAGAGGGAGGAGAGGGA
                     GGAGGAGAGGGAGAGGGAGGAGAGGGAGGAGGAGAGGGAGAGGAGGAGGAGAGGAGAGG
                     GAGGAGGAGAGGAGAGGAGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGGAGAG
                     GGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGAGGAGAGGGGRGRGGSGGRGRG
                     GSGGRGRGGSGGRRGRGRERARGGSRERARGRGRGRGEKRPRSPSSQSSSSGSPPRRPP
                     PGRRPFFHPVGEADYFEYHQEGGPDGEPDVPPGAIEQGPADDPGEGPSTGPRGQGDGGR
                     RKKGGWFGKHRGQGGSNPKFENIAEGLRALLARSHVERTTDEGTWVAGVFVYGGSKTSL
                     YNLRRGTALAIPQCRLTPLSRLPFGMAPGPGPQPGPLRESIVCYFMVFLQTHIFAEVLK
                     DAIKDLVMTKPAPTCNIRVTVCSFDDGVDLPPWFPPMVEGAAAEGDDGDDGDEGGDGDE
                     GEEGQE"
     protein_bind    10667..10700
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      10887..12676
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
     promoter        13147..13460
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     13379..13399
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     complement(13611..13628)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13782..14370)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13862..13881)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14541..15332,15333..15401))
                     /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     15164..15183
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15402..15506)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
      541 ggggcgggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcctcgaccg
     1741 gtgccaccat gtacccatac gatgttccag attacgcttc gccgaagaaa aagcgcaagg
     1801 tcgaagcgtc cgacaagaag tacagcatcg gcctggccat cggcaccaac tctgtgggct
     1861 gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca
     1921 ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa
     1981 cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc
     2041 ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct
     2101 tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca
     2161 tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc
     2221 tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc
     2281 tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca
     2341 acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg
     2401 aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca
     2461 agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt
     2521 tcggcaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc
     2581 tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc
     2641 tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg
     2701 acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga
     2761 gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc
     2821 tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg
     2881 gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc
     2941 ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc
     3001 tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag
     3061 agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg
     3121 aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg
     3181 gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact
     3241 tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact
     3301 tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact
     3361 tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg
     3421 ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga
     3481 aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg
     3541 tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc
     3601 tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag
     3661 atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa
     3721 cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg
     3781 gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga
     3841 caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc
     3901 acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg
     3961 atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc
     4021 tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga
     4081 acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc
     4141 gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag
     4201 aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga
     4261 atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg
     4321 tcgacgccat cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga
     4381 ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga
     4441 agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg
     4501 acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca
     4561 agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc
     4621 ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc
     4681 tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga
     4741 tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga
     4801 tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg
     4861 tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct
     4921 tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc
     4981 ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc
     5041 gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga
     5101 ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata
     5161 agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca
     5221 ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga
     5281 agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc
     5341 ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc
     5401 tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg
     5461 gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc
     5521 tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt
     5581 ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca
     5641 agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc
     5701 gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc
     5761 tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca
     5821 gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga
     5881 cacggatcga cctgtctcag ctgggaggcg acagccccaa gaagaagaga aaggtggagg
     5941 ccagcctcga gggaatcgat atggtgagca agggcgagga gctgttcacc ggggtggtgc
     6001 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     6061 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     6121 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     6181 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     6241 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     6301 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     6361 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     6421 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     6481 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     6541 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     6601 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     6661 tggacgagct gtacaagtaa tttaaagaat tcaagcttcg gggactagtc atatgataat
     6721 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     6781 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     6841 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     6901 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     6961 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     7021 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     7081 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     7141 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     7201 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     7261 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg gtaaattcac
     7321 tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca
     7381 caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct
     7441 tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     7501 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa
     7561 tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg
     7621 ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca
     7681 tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc
     7741 tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg
     7801 actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa
     7861 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     7921 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     7981 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     8041 agcggatctc aattccgatc atattcaata acccttaata taacttcgta taatgtatgc
     8101 tatacgaagt tattaggtct gaagaggagt ttacgtccag ccaagcttag gatcaattct
     8161 catgtttgac agcttatcat cgataagctg atcctcacag gccgcaccca gcttttcttc
     8221 cgttgcccca gtagcatctc tgtctggtga ccttgaagag gaagaggagg ggtcccgaga
     8281 atccccatcc ctaccgtcca gcaaaaaggg ggacgaggaa tttgaggcct ggcttgaggc
     8341 tcaggacgca aatcttgagg atgttcagcg ggagttttcc gggctgcgag taattggtga
     8401 tgaggacgag gatggttcgg aggatgggga attttcagac ctggatctgt ctgacagcga
     8461 ccatgaaggg gatgagggtg ggggggctgt tggagggggc aggagtctgc actccctgta
     8521 ttcactgagc gtcgtctaat aaagatgtct attgatctct tttagtgtga atcatgtctg
     8581 acgaggggcc aggtacagga cctggaaatg gcctaggaga gaagggagac acatctggac
     8641 cagaaggctc cggcggcagt ggacctcaaa gaagaggggg tgataaccat ggacgaggac
     8701 ggggaagagg acgaggacga ggaggcggaa gaccaggagc cccgggcggc tcaggatcag
     8761 ggccaagaca tagagatggt gtccggagac cccaaaaacg tccaagttgc attggctgca
     8821 aagggaccca cggtggaaca ggagcaggag caggagcggg aggggcagga gcaggagggg
     8881 caggagcagg aggaggggca ggagcaggag gaggggcagg aggggcagga ggggcaggag
     8941 gggcaggagc aggaggaggg gcaggagcag gaggaggggc aggaggggca ggaggggcag
     9001 gagcaggagg aggggcagga gcaggaggag gggcaggagg ggcaggagca ggaggagggg
     9061 caggaggggc aggaggggca ggagcaggag gaggggcagg agcaggagga ggggcaggag
     9121 gggcaggagc aggaggaggg gcaggagggg caggaggggc aggagcagga ggaggggcag
     9181 gagcaggagg ggcaggaggg gcaggagggg caggagcagg aggggcagga gcaggaggag
     9241 gggcaggagg ggcaggaggg gcaggagcag gaggggcagg agcaggaggg gcaggagcag
     9301 gaggggcagg agcaggaggg gcaggagggg caggagcagg aggggcagga ggggcaggag
     9361 caggaggggc aggaggggca ggagcaggag gaggggcagg aggggcagga gcaggaggag
     9421 gggcaggagg ggcaggagca ggaggggcag gaggggcagg agcaggaggg gcaggagggg
     9481 caggagcagg aggggcagga ggggcaggag caggaggagg ggcaggagca ggaggggcag
     9541 gagcaggagg tggaggccgg ggtcgaggag gcagtggagg ccggggtcga ggaggtagtg
     9601 gaggccgggg tcgaggaggt agtggaggcc gccggggtag aggacgtgaa agagccaggg
     9661 ggggaagtcg tgaaagagcc agggggagag gtcgtggacg tggagaaaag aggcccagga
     9721 gtcccagtag tcagtcatca tcatccgggt ctccaccgcg caggccccct ccaggtagaa
     9781 ggccattttt ccaccctgta ggggaagccg attattttga ataccaccaa gaaggtggcc
     9841 cagatggtga gcctgacgtg cccccgggag cgatagagca gggccccgca gatgacccag
     9901 gagaaggccc aagcactgga ccccggggtc agggtgatgg aggcaggcgc aaaaaaggag
     9961 ggtggtttgg aaagcatcgt ggtcaaggag gttccaaccc gaaatttgag aacattgcag
    10021 aaggtttaag agctctcctg gctaggagtc acgtagaaag gactaccgac gaaggaactt
    10081 gggtcgccgg tgtgttcgta tatggaggta gtaagacctc cctttacaac ctaaggcgag
    10141 gaactgccct tgctattcca caatgtcgtc ttacaccatt gagtcgtctc ccctttggaa
    10201 tggcccctgg acccggccca caacctggcc cgctaaggga gtccattgtc tgttatttca
    10261 tggtcttttt acaaactcat atatttgctg aggttttgaa ggatgcgatt aaggaccttg
    10321 ttatgacaaa gcccgctcct acctgcaata tcagggtgac tgtgtgcagc tttgacgatg
    10381 gagtagattt gcctccctgg tttccaccta tggtggaagg ggctgccgcg gagggtgatg
    10441 acggagatga cggagatgaa ggaggtgatg gagatgaggg tgaggaaggg caggagtgat
    10501 gtaacttgtt aggagacgcc ctcaatcgta ttaaaagccg tgtattcccc cgcactaaag
    10561 aataaatccc cagtagacat catgcgtgct gttggtgtat ttctggccat ctgtcttgtc
    10621 accattttcg tcctcccaac atggggcaat tgccggaacc cttaatataa cttcgtataa
    10681 tgtatgctat acgaagttat taggtccctc gaagaggttc actagcggat ctcaattggg
    10741 catacccatg ttgtcacgtc actcagctcc gcgctcaaca ccttctcgcg ttggaaaaca
    10801 ttagcgacat ttacctggtg agcaatcaga catgcgacgg ctttagcctg gcctccttaa
    10861 attcacctaa gaatgggagc aaccagcagg aaaaggacaa gcagcgaaaa ttcacgcccc
    10921 cttgggaggt ggcggcatat gcaaaggata gcactcccac tctactactg ggtatcatat
    10981 gctgactgta tatgcatgag gatagcatat gctacccgga tacagattag gatagcatat
    11041 actacccaga tatagattag gatagcatat gctacccaga tatagattag gatagcctat
    11101 gctacccaga tataaattag gatagcatat actacccaga tatagattag gatagcatat
    11161 gctacccaga tatagattag gatagcctat gctacccaga tatagattag gatagcatat
    11221 gctacccaga tatagattag gatagcatat gctatccaga tatttgggta gtatatgcta
    11281 cccagatata aattaggata gcatatacta ccctaatctc tattaggata gcatatgcta
    11341 cccggataca gattaggata gcatatacta cccagatata gattaggata gcatatgcta
    11401 cccagatata gattaggata gcctatgcta cccagatata aattaggata gcatatacta
    11461 cccagatata gattaggata gcatatgcta cccagatata gattaggata gcctatgcta
    11521 cccagatata gattaggata gcatatgcta tccagatatt tgggtagtat atgctaccca
    11581 tggcaacatt agcccaccgt gctctcagcg acctcgtgaa tatgaggacc aacaaccctg
    11641 tgcttggcgc tcaggcgcaa gtgtgtgtaa tttgtcctcc agatcgcagc aatcgcgccc
    11701 ctatcttggc ccgcccacct acttatgcag gtattccccg gggtgccatt agtggttttg
    11761 tgggcaagtg gtttgaccgc agtggttagc ggggttacaa tcagccaagt tattacaccc
    11821 ttattttaca gtccaaaacc gcagggcggc gtgtgggggc tgacgcgtgc ccccactcca
    11881 caatttcaaa aaaaagagtg gccacttgtc tttgtttatg ggccccattg gcgtggagcc
    11941 ccgtttaatt ttcgggggtg ttagagacaa ccagtggagt ccgctgctgt cggcgtccac
    12001 tctctttccc cttgttacaa atagagtgta acaacatggt tcacctgtct tggtccctgc
    12061 ctgggacaca tcttaataac cccagtatca tattgcacta ggattatgtg ttgcccatag
    12121 ccataaattc gtgtgagatg gacatccagt ctttacggct tgtccccacc ccatggattt
    12181 ctattgttaa agatattcag aatgtttcat tcctacacta gtatttattg cccaaggggt
    12241 ttgtgagggt tatattggtg tcatagcaca atgccaccac tgaacccccc gtccaaattt
    12301 tattctgggg gcgtcacctg aaaccttgtt ttcgagcacc tcacatacac cttactgttc
    12361 acaactcagc agttattcta ttagctaaac gaaggagaat gaagaagcag gcgaagattc
    12421 aggagagttc actgcccgct ccttgatctt cagccactgc ccttgtgact aaaatggttc
    12481 actaccctcg tggaatcctg accccatgta aataaaaccg tgacagctca tggggtggga
    12541 gatatcgctg ttccttagga cccttttact aaccctaatt cgatagcata tgcttcccgt
    12601 tgggtaacat atgctattga attagggtta gtctggatag tatatactac tacccgggaa
    12661 gcatatgcta cccgtttagg gttaacaagg gggccttata aacactattg ctaatgccct
    12721 cttgagggtc cgcttatcgg tagctacaca ggcccctctg attgacgttg gtgtagcctc
    12781 ccgtagtctt cctgggcccc tgggaggtac atgtccccca gcattggtgt aagagcttca
    12841 gccaagagtt acacataaag gcaatgttgt gttgcagtcc acagactgca aagtctgctc
    12901 caggatgaaa gccactcagt gttggcaaat gtgcacatcc atttataagg atgtcaacta
    12961 cagtcagaga acccctttgt gtttggtccc cccccgtgtc acatgtggaa cagggcccag
    13021 ttggcaagtt gtaccaacca actgaaggga ttacatgcac tgccccgcga agaaggggca
    13081 gagatgtcgt agtcaggttt agttcgtccg gggcggggca tcgatcctct agagtcgacg
    13141 ctagcggatc cgacgccgcc atctctaggc ccgcgccggc cccctcgcac agacttgtgg
    13201 gagaagctcg gctactcccc tgccccggtt aatttgcata taatatttcc tagtaactat
    13261 agaggcttaa tgtgcgataa aagacagata atctgttctt tttaatacta gctacatttt
    13321 acatgatagg cttggatttc tataagagat acaaatacta aattattatt ttaaaaaaca
    13381 gcacaaaagg aaactcaccc taactgtaaa gtaattgtgt gttttgagac tataaatatc
    13441 ccttggagaa aagccttgtt tgggccgact ccagtggtaa tctacttcaa gagagtagat
    13501 taccactgga gtcttttttg gaattcctgc agcccggggg atccgctgca ttaatgaatc
    13561 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
    13621 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
    13681 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
    13741 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
    13801 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
    13861 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
    13921 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
    13981 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
    14041 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
    14101 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
    14161 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
    14221 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
    14281 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
    14341 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
    14401 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
    14461 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
    14521 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
    14581 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
    14641 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
    14701 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
    14761 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
    14821 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
    14881 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
    14941 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
    15001 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
    15061 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
    15121 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
    15181 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
    15241 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
    15301 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
    15361 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
    15421 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
    15481 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgg
//
