LOCUS       Exported               15848 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Episomal plasmid encoding dCas9VP192.
ACCESSION   .
VERSION     .
KEYWORDS    pCXLE-dCas9VP192-T2A-EGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15848)
  AUTHORS   Balboa D, Weltner J, Eurola S, Trokovic R, Wartiovaara K, Otonkoski 
            T
  TITLE     Conditionally Stabilized dCas9 Activator for Controlling Gene 
            Expression in Human Cell Reprogramming and Differentiation.
  JOURNAL   Stem Cell Reports. 2015 Sep 8;5(3):448-59. doi: 
            10.1016/j.stemcr.2015.08.001.
  PUBMED    26352799
REFERENCE   2  (bases 1 to 15848)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..15848
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             1812..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             5973..6083
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     CDS             6135..6245
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     CDS             6297..6407
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     CDS             6459..6569
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     CDS             6609..6662
                     /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"
     CDS             6669..7388
                     /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             6669..7385
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6714..6735)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6975..6994)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7322..7343
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    7426..8014
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7479..7499)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7897..7908)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8104..8123)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    8169..8224
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(8223..8242)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(8585..8601)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8585..8601)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8598..8620)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8609..8625
                     /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(8633..8663)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8678..8699
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    complement(8788..8821)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             9282..11207
                     /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    complement(11375..11408)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     rep_origin      11595..13384
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
     primer_bind     complement(13926..13943)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14097..14685)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14177..14196)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(14856..15716)
                     /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     15479..15498
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15717..15821)
                     /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 gtgcgaaagc ggcgggccgg ccgacgccct tgacgatttt gacttagaca
     6001 tgctcccagc cgatgccctt gacgactttg accttgatat gctgcctgct gacgctcttg
     6061 acgattttga ccttgacatg ctccccgggt cactacccag cgctagcgtc gagtgcgaaa
     6121 gcggcgggcc ggccgacgcc cttgacgatt ttgacttaga catgctccca gccgatgccc
     6181 ttgacgactt tgaccttgat atgctgcctg ctgacgctct tgacgatttt gaccttgaca
     6241 tgctccccgg gtcactaccc agcgctagcg tcgagtgcga aagcggcggg ccggccgacg
     6301 cccttgacga ttttgactta gacatgctcc cagccgatgc ccttgacgac tttgaccttg
     6361 atatgctgcc tgctgacgct cttgacgatt ttgaccttga catgctcccc gggtcactac
     6421 ccagcgctag cgtcgagtgc gaaagcggcg ggccggccga cgcccttgac gattttgact
     6481 tagacatgct cccagccgat gcccttgacg actttgacct tgatatgctg cctgctgacg
     6541 ctcttgacga ttttgacctt gacatgctcc ccgggtcact acccagcgct agcgtcgacg
     6601 gctccggaga gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc
     6661 caatcgatat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     6721 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     6781 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     6841 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     6901 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     6961 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     7021 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     7081 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     7141 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     7201 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     7261 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     7321 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     7381 acaagtaatt taaagaattc aagcttcggg gactagtcat atgataatca acctctggat
     7441 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     7501 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     7561 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     7621 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     7681 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     7741 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     7801 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     7861 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     7921 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     7981 acgagtcgga tctccctttg ggccgcctcc ccgcatcggt aaattcactc ctcaggtgca
     8041 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg
     8101 agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac
     8161 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     8221 ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt
     8281 ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag
     8341 aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt
     8401 gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct
     8461 aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt
     8521 catagctgtc cctcttctct tatggagatc cctcgacctg cagcccaagc ttggcgtaat
     8581 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     8641 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     8701 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatctcaa
     8761 ttccgatcat attcaataac ccttaatata acttcgtata atgtatgcta tacgaagtta
     8821 ttaggtctga agaggagttt acgtccagcc aagcttagga tcaattctca tgtttgacag
     8881 cttatcatcg ataagctgat cctcacaggc cgcacccagc ttttcttccg ttgccccagt
     8941 agcatctctg tctggtgacc ttgaagagga agaggagggg tcccgagaat ccccatccct
     9001 accgtccagc aaaaaggggg acgaggaatt tgaggcctgg cttgaggctc aggacgcaaa
     9061 tcttgaggat gttcagcggg agttttccgg gctgcgagta attggtgatg aggacgagga
     9121 tggttcggag gatggggaat tttcagacct ggatctgtct gacagcgacc atgaagggga
     9181 tgagggtggg ggggctgttg gagggggcag gagtctgcac tccctgtatt cactgagcgt
     9241 cgtctaataa agatgtctat tgatctcttt tagtgtgaat catgtctgac gaggggccag
     9301 gtacaggacc tggaaatggc ctaggagaga agggagacac atctggacca gaaggctccg
     9361 gcggcagtgg acctcaaaga agagggggtg ataaccatgg acgaggacgg ggaagaggac
     9421 gaggacgagg aggcggaaga ccaggagccc cgggcggctc aggatcaggg ccaagacata
     9481 gagatggtgt ccggagaccc caaaaacgtc caagttgcat tggctgcaaa gggacccacg
     9541 gtggaacagg agcaggagca ggagcgggag gggcaggagc aggaggggca ggagcaggag
     9601 gaggggcagg agcaggagga ggggcaggag gggcaggagg ggcaggaggg gcaggagcag
     9661 gaggaggggc aggagcagga ggaggggcag gaggggcagg aggggcagga gcaggaggag
     9721 gggcaggagc aggaggaggg gcaggagggg caggagcagg aggaggggca ggaggggcag
     9781 gaggggcagg agcaggagga ggggcaggag caggaggagg ggcaggaggg gcaggagcag
     9841 gaggaggggc aggaggggca ggaggggcag gagcaggagg aggggcagga gcaggagggg
     9901 caggaggggc aggaggggca ggagcaggag gggcaggagc aggaggaggg gcaggagggg
     9961 caggaggggc aggagcagga ggggcaggag caggaggggc aggagcagga ggggcaggag
    10021 caggaggggc aggaggggca ggagcaggag gggcaggagg ggcaggagca ggaggggcag
    10081 gaggggcagg agcaggagga ggggcaggag gggcaggagc aggaggaggg gcaggagggg
    10141 caggagcagg aggggcagga ggggcaggag caggaggggc aggaggggca ggagcaggag
    10201 gggcaggagg ggcaggagca ggaggagggg caggagcagg aggggcagga gcaggaggtg
    10261 gaggccgggg tcgaggaggc agtggaggcc ggggtcgagg aggtagtgga ggccggggtc
    10321 gaggaggtag tggaggccgc cggggtagag gacgtgaaag agccaggggg ggaagtcgtg
    10381 aaagagccag ggggagaggt cgtggacgtg gagaaaagag gcccaggagt cccagtagtc
    10441 agtcatcatc atccgggtct ccaccgcgca ggccccctcc aggtagaagg ccatttttcc
    10501 accctgtagg ggaagccgat tattttgaat accaccaaga aggtggccca gatggtgagc
    10561 ctgacgtgcc cccgggagcg atagagcagg gccccgcaga tgacccagga gaaggcccaa
    10621 gcactggacc ccggggtcag ggtgatggag gcaggcgcaa aaaaggaggg tggtttggaa
    10681 agcatcgtgg tcaaggaggt tccaacccga aatttgagaa cattgcagaa ggtttaagag
    10741 ctctcctggc taggagtcac gtagaaagga ctaccgacga aggaacttgg gtcgccggtg
    10801 tgttcgtata tggaggtagt aagacctccc tttacaacct aaggcgagga actgcccttg
    10861 ctattccaca atgtcgtctt acaccattga gtcgtctccc ctttggaatg gcccctggac
    10921 ccggcccaca acctggcccg ctaagggagt ccattgtctg ttatttcatg gtctttttac
    10981 aaactcatat atttgctgag gttttgaagg atgcgattaa ggaccttgtt atgacaaagc
    11041 ccgctcctac ctgcaatatc agggtgactg tgtgcagctt tgacgatgga gtagatttgc
    11101 ctccctggtt tccacctatg gtggaagggg ctgccgcgga gggtgatgac ggagatgacg
    11161 gagatgaagg aggtgatgga gatgagggtg aggaagggca ggagtgatgt aacttgttag
    11221 gagacgccct caatcgtatt aaaagccgtg tattcccccg cactaaagaa taaatcccca
    11281 gtagacatca tgcgtgctgt tggtgtattt ctggccatct gtcttgtcac cattttcgtc
    11341 ctcccaacat ggggcaattg ccggaaccct taatataact tcgtataatg tatgctatac
    11401 gaagttatta ggtccctcga agaggttcac tagcggatct caattgggca tacccatgtt
    11461 gtcacgtcac tcagctccgc gctcaacacc ttctcgcgtt ggaaaacatt agcgacattt
    11521 acctggtgag caatcagaca tgcgacggct ttagcctggc ctccttaaat tcacctaaga
    11581 atgggagcaa ccagcaggaa aaggacaagc agcgaaaatt cacgccccct tgggaggtgg
    11641 cggcatatgc aaaggatagc actcccactc tactactggg tatcatatgc tgactgtata
    11701 tgcatgagga tagcatatgc tacccggata cagattagga tagcatatac tacccagata
    11761 tagattagga tagcatatgc tacccagata tagattagga tagcctatgc tacccagata
    11821 taaattagga tagcatatac tacccagata tagattagga tagcatatgc tacccagata
    11881 tagattagga tagcctatgc tacccagata tagattagga tagcatatgc tacccagata
    11941 tagattagga tagcatatgc tatccagata tttgggtagt atatgctacc cagatataaa
    12001 ttaggatagc atatactacc ctaatctcta ttaggatagc atatgctacc cggatacaga
    12061 ttaggatagc atatactacc cagatataga ttaggatagc atatgctacc cagatataga
    12121 ttaggatagc ctatgctacc cagatataaa ttaggatagc atatactacc cagatataga
    12181 ttaggatagc atatgctacc cagatataga ttaggatagc ctatgctacc cagatataga
    12241 ttaggatagc atatgctatc cagatatttg ggtagtatat gctacccatg gcaacattag
    12301 cccaccgtgc tctcagcgac ctcgtgaata tgaggaccaa caaccctgtg cttggcgctc
    12361 aggcgcaagt gtgtgtaatt tgtcctccag atcgcagcaa tcgcgcccct atcttggccc
    12421 gcccacctac ttatgcaggt attccccggg gtgccattag tggttttgtg ggcaagtggt
    12481 ttgaccgcag tggttagcgg ggttacaatc agccaagtta ttacaccctt attttacagt
    12541 ccaaaaccgc agggcggcgt gtgggggctg acgcgtgccc ccactccaca atttcaaaaa
    12601 aaagagtggc cacttgtctt tgtttatggg ccccattggc gtggagcccc gtttaatttt
    12661 cgggggtgtt agagacaacc agtggagtcc gctgctgtcg gcgtccactc tctttcccct
    12721 tgttacaaat agagtgtaac aacatggttc acctgtcttg gtccctgcct gggacacatc
    12781 ttaataaccc cagtatcata ttgcactagg attatgtgtt gcccatagcc ataaattcgt
    12841 gtgagatgga catccagtct ttacggcttg tccccacccc atggatttct attgttaaag
    12901 atattcagaa tgtttcattc ctacactagt atttattgcc caaggggttt gtgagggtta
    12961 tattggtgtc atagcacaat gccaccactg aaccccccgt ccaaatttta ttctgggggc
    13021 gtcacctgaa accttgtttt cgagcacctc acatacacct tactgttcac aactcagcag
    13081 ttattctatt agctaaacga aggagaatga agaagcaggc gaagattcag gagagttcac
    13141 tgcccgctcc ttgatcttca gccactgccc ttgtgactaa aatggttcac taccctcgtg
    13201 gaatcctgac cccatgtaaa taaaaccgtg acagctcatg gggtgggaga tatcgctgtt
    13261 ccttaggacc cttttactaa ccctaattcg atagcatatg cttcccgttg ggtaacatat
    13321 gctattgaat tagggttagt ctggatagta tatactacta cccgggaagc atatgctacc
    13381 cgtttagggt taacaagggg gccttataaa cactattgct aatgccctct tgagggtccg
    13441 cttatcggta gctacacagg cccctctgat tgacgttggt gtagcctccc gtagtcttcc
    13501 tgggcccctg ggaggtacat gtcccccagc attggtgtaa gagcttcagc caagagttac
    13561 acataaaggc aatgttgtgt tgcagtccac agactgcaaa gtctgctcca ggatgaaagc
    13621 cactcagtgt tggcaaatgt gcacatccat ttataaggat gtcaactaca gtcagagaac
    13681 ccctttgtgt ttggtccccc cccgtgtcac atgtggaaca gggcccagtt ggcaagttgt
    13741 accaaccaac tgaagggatt acatgcactg ccccgcgaag aaggggcaga gatgtcgtag
    13801 tcaggtttag ttcgtccggg gcggggcatc gatcctctag agtcgacgct agcggatccg
    13861 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
    13921 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
    13981 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
    14041 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
    14101 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
    14161 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
    14221 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
    14281 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
    14341 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
    14401 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
    14461 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
    14521 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
    14581 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
    14641 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
    14701 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
    14761 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
    14821 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
    14881 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
    14941 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
    15001 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
    15061 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
    15121 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
    15181 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
    15241 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
    15301 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
    15361 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
    15421 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
    15481 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
    15541 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
    15601 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
    15661 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
    15721 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
    15781 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
    15841 ccacctgg
//
