LOCUS       Exported               13567 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Donor plasmid with a Cas9-p300 expression cassette, a M2rtTA 
            expression cassette, a T2A-puro selection cassette, and two gRNA 
            recognition sequences at both ends of the whole insertion DNA 
            fragment..
ACCESSION   .
VERSION     .
KEYWORDS    Pblu-AAVS1-Cas9-p300-M2rtTA-AAVS1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13567)
  AUTHORS   Ma S, Lv J, Sun J, Tang P, Li H, Zhou H, Zhang Z, Lin Y, Rong Z
  TITLE     iKA-CRISPR hESCs for inducible and multiplex orthogonal gene 
            knockout and activation.
  JOURNAL   FEBS Lett. 2018 Jun 5. doi: 10.1002/1873-3468.13127.
  PUBMED    29869798
REFERENCE   2  (bases 1 to 13567)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..13567
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(6..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(143..164)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     355..374
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    706..731
                     /label=SA
                     /note="splice acceptor site"
     CDS             755..808
                     /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             818..1417
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(818..837)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1314..1334
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(1449..1466)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1455..1679
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    1686..1767
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     1711..1730
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1782..2528)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(2601..2620)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          2628..3644
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(2682..2704)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3645..3922)
                     /label=chicken beta-actin promoter
     enhancer        3924..4227
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    4325..4786
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     CDS             complement(4811..6661)
                     /codon_start=1
                     /gene="EP300"
                     /product="catalytic histone acetyltransferase core of the 
                     human E1A-associated protein p300 (Delvecchio et al., 
                     2013)"
                     /label=p300 core
                     /translation="IFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDI
                     VKSPMDLSTIKRKLDTGQYQEPWQYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQ
                     EIDPVMQSLGYCCGRKLEFSPQTLCCYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQG
                     ESVSLGDDPSQPQTTINKEQFSKRKNDTLDPELFVECTECGRKMHQICVLHHEIIWPAG
                     FVCDGCLKKSARTRKENKFSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHA
                     SDKTVEVKPGMKARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPP
                     PNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDY
                     IFHCHPPDQKIPKPKRLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFE
                     GDFWPNVLEESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLS
                     RGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDL
                     MDGRDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQD"
     regulatory      5038..5047
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             complement(6665..6685)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(6698..10801)
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(10823..10843)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(10850..10915)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(10952..10976)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(10993..11307)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     primer_bind     complement(10996..11020)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    11067..11085
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11103..11121
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11138..11156
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11174..11192
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11210..11228
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11245..11263
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    11281..11299
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(11382..11402)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(11382..11400)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11421..11437)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11421..11437)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11434..11456)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11445..11461
                     /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(11469..11499)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11514..11535
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11652..11669)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11823..12411)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11903..11922)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(12582..13442)
                     /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     13205..13224
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13443..13547)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
       61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac
      661 cgggcccccc ctcgaggtca ccaatcctgt ccctagtgga agcttctgac ctcttctctt
      721 cctcccacag ggcctcgaga gatctggcag cggagagggc agaggaagtc ttctaacatg
      781 cggtgacgtg gaggagaatc ccggccctag gctcgagatg accgagtaca agcccacggt
      841 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
      901 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
      961 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     1021 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     1081 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     1141 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     1201 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     1261 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     1321 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     1381 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgatct agagggcccg tttaaacccg
     1441 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     1501 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     1561 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     1621 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggg
     1681 tcgaggcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     1741 accattataa gctgcaataa acaagttaac aacaacaatt gttacccggg gagcatgtca
     1801 aggtcaaaat cgtcaagagc gtcagcaggc agcatatcaa ggtcaaagtc gtcaagggca
     1861 tcggctggga gcatgtctaa gtcaaaatcg tcaagggcgt cggccggccc gccgctttcg
     1921 cactttagct gtttctccag gccacatatg attagttcca ggccgaaaag gaaggcaggt
     1981 tcggctccct gccggtcgaa cagctcaatt gcttgtctca gaagtggggg catagaatcg
     2041 gtggtaggtg tctctctttc ctcttttgct acttgatgct cctgttcctc caatacgcag
     2101 cccagtgtaa agtggcccac ggcggacaga gcgtacagtg cgttctccag ggagaagcct
     2161 tgctgacaca ggaacgcgag ctgattttcc agggtttcgt actgtttctc tgttgggcgg
     2221 gtgccgagat gcactttagc cccgtcgcga tgtgagagga gagcacagcg gaatgacttg
     2281 gcgttgttcc gcagaaagtc ttgccatgac tcgccttcca gggggcagaa gtgggtatga
     2341 tgcctgtcca gcatctcgat tggcagggca tcgagcaggg cccgcttgtt cttcacgtgc
     2401 cagtacaggg taggctgctc aactcccagc ttttgagcga gtttccttgt cgtcaggcct
     2461 tcgataccga ctccattgag taattccaga gcgccgttta tgactttgct cttgtccagt
     2521 ctagacatgg tgaattcctg cagcccgggg gatccactag cttcgatcaa gccgaattct
     2581 ttgccaaaat gatgagacag cacaataacc agcacgttgc ccaggagctg taggaaaaag
     2641 aagaaggcat gaacatggtt agcagaggct ctagagccgc cggtcacacg ccagaagccg
     2701 aaccccgccc tgccccgtcc cccccgaagg cagccgtccc cccgcggaca gccccgaggc
     2761 tggagaggga gaaggggacg gcggcgcggc gacgcacgaa ggccctcccc gcccatttcc
     2821 ttcctgccgg cgccgcaccg cttcgccccg cgcccgctag agggggtgcg gcggcgcctc
     2881 ccagatttcg gctccgcaca gatttgggac aaaggaagtc cctgcgccct ctcgcacgat
     2941 taccataaaa ggcaatggct gcggctcgcc gcgcctcgac agccgccggc gctctggggg
     3001 ccgccgcgcc cctcccccga gccctccccg gcccgaggcg gccccgcccc gcccggcacc
     3061 cccacctgcc gccacccccc gcccggcacg gcgagccccg cgccacgccc cgtacggagc
     3121 cccgcacccg aagccgggcc gtgctcagca actcggggag gggggtgcag ggggggttgc
     3181 agcccgaccg acgcgcccac accccctgct caccccccca cgcacacacc ccgcacgcag
     3241 cctttgttcc cctcgcagcc ccccccgcac cgcggggcac cgcccccggc cgcgctcccc
     3301 tcgcgcacac tgcggagcgc acaaagcccc gcgccgcgcc cgcagcgctc acagccgccg
     3361 ggcagcgcgg agccgcacgc ggcgctcccc acgcacacac acacgcacgc accccccgag
     3421 ccgctccccc cgcacaaagg gccctcccgg agcccctcaa ggctttcacg cagccacaga
     3481 aaagaaacaa gccgtcatta aaccaagcgc taattacagc ccggaggaga agggccgtcc
     3541 cgcccgctca cctgtgggag taacgcggtc agtcagagcc ggggcgggcg gcgcgaggcg
     3601 gcggcggagc ggggcacggg gcgaaggcag cgcgcagcga ctcccgcccg ccgcgcgctt
     3661 cgctttttat agggccgccg ccgccgccgc ctcgccataa aaggaaactt tcggagcgcg
     3721 ccgctctgat tggctgccgc cgcacctctc cgcctcgccc cgccccgccc ctcgccccgc
     3781 cccgccccgc ctggcgcgcg cccccccccc ccccccgccc ccatcgctgc acaaaataat
     3841 taaaaaataa ataaatacaa aattgggggt ggggaggggg gggagatggg gagagtgaag
     3901 cagaacgtgg ggctcacctc gaccatggta atagcgatga ctaatacgta gatgtactgc
     3961 caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg
     4021 tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg
     4081 gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact
     4141 atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc
     4201 gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac
     4261 cccgtaattg attactatta ataactagtt ctagagcggc cgccaccgcg gtggagctcc
     4321 cagcttctga tggaattaga acttggcaaa acaatactga gaatgaagtg tatgtggaac
     4381 agaggctgct gatctcgttc ttcaggctat gaaactgaca catttggaaa ccacagtact
     4441 tagaaccaca aagtgggaat caagagaaaa acaatgatcc cacgagagat ctatagatct
     4501 atagatcatg agtgggagga atgagctggc ccttaatttg gttttgcttg tttaaattat
     4561 gatatccaac tatgaaacat tatcataaag caatagtaaa gagccttcag taaagagcag
     4621 gcatttatct aatcccaccc cacccccacc cccgtagctc caatccttcc attcaaaatg
     4681 taggtactct gttctcaccc ttcttaacaa agtatgacag gaaaaacttc cattttagtg
     4741 gacatcttta ttgtttaata gatcatcaat ttctgcagac ttacagcgga tccccacgcg
     4801 tttaaccggt gtcctggctc tgcgtgtgca gctccaccag catgcacatg gtggaccact
     4861 gggctcttcg gagtgaagag aactccaggt gcttgtccct tgccagcgtg agaaacgcat
     4921 cccgaccatc catcagatcg caggggatga gaggatcagg atcaacaatg ggaggcaggg
     4981 agttggcagc agggccagca atgaggcgga tcacaaagaa gacctcttta tgcttctcca
     5041 tggtggcata tagtttctgt gagaggtcgt tagatacatt gggcatcccg ggtttcttct
     5101 tgttgcccct actcaggctg ctcttatttt tgctggtttt cttattattc ttctttttag
     5161 catttttgct gtctcccttg gtcacatctg tgctttcatt gctggtgttt tcctctcgtt
     5221 ttctctcttc ttcctcctgt tccagttcct taatgctttc ttccagaaca ttgggccaga
     5281 aatcaccctc gaaataaggc aattcctttg cacttgttaa tctatcttca gtagcttgtt
     5341 taaaaatatc cttgtagtca tggacaatac gctctgatac agccttgtca agcatttttt
     5401 tgtaccattc ctgcagtcgc ttgggcttgg gtatcttctg gtcaggagga tggcaatgga
     5461 agatataatc atctccctca cttggtggac atgcccaaat atgccctgtt gtgtaaccta
     5521 atttcttgac atattctaaa tatccaatta ggatttcatg atagactgca gtcctcaagc
     5581 atttaggacg gaagaaatga acactatcga ggtaagatat gtatactctc ctctggttgg
     5641 gtggagggca gtcagagcca tactcttgaa catgcatgcc aaagaagcac aggtcaacac
     5701 catcaatttc ttcaaaggca aagagggctt tggttcggta tggaaaggat tctgccatct
     5761 ctccactgtc cacaaacctt gctttcatgc ctggttttac ttccacggtt ttgtcagaag
     5821 catgaactac tctaacagtg acctctcctg actcagggtg attctgtcgc ctcagaaagt
     5881 cattcacacg attctctaga aaggtgccaa gtctggtaga tggcaacctt ttagcagaaa
     5941 acttattttc tttcctagtt cgtgcacttt tctttaaaca gccatcacag acgaatccag
     6001 caggccagat gatctcatgg tgaaggacac agatctgatg catctttctt ccgcactctg
     6061 tacattcaac aaacagttca ggatccagtg tgtcattttt tctcttggaa aattgttctt
     6121 tatttattgt agtttgaggc tgggaagggt catcccccaa agaaacgctc tccccttgga
     6181 tctcattgaa acacttctca cagaaatgat acctgttctg gtaactgtaa taagtggcat
     6241 cacgaggtat tgtgcacaac tgtttgccgt agcaacacag tgtctgtgga gagaactcca
     6301 actttctgcc gcaacaatat ccaaggcttt gcatcactgg gtcaatttct tgttcaaaga
     6361 cctcagagag cttggagcag tatttgtata cccgtgatgt tttccggtta tataaccagg
     6421 cattattgaa cataagccaa atatcatcga catactgcca gggctcctga tactgtccag
     6481 tgtctaactt cctcttaatg gtagaaagat ccatggggct cttcacaata tcaaagtaat
     6541 cagggattcc taaaagctga gggtccacag gttgacgaaa gggaagggat tctggatcct
     6601 gacggtaaag tgcctccaat gttggcatca gtgcctgtcg tagttcttct ggtttgaaaa
     6661 tgctcacctt cctcttcttc ttggggtcag ccctgctgtc tccaccgagc tgagagaggt
     6721 cgattcttgt ttcatagagc cccgtaattg actgatgaat cagtgtggcg tccaggacct
     6781 cctttgtaga ggtgtaccgc tttctgtcta tggtggtgtc gaagtacttg aaggctgcag
     6841 gcgcgcccaa gttggtcaga gtaaacaagt ggataatgtt ttctgcctgc tccctgatgg
     6901 gcttatccct gtgcttattg taagcagaaa gcaccttatc gaggttagcg tcggcgagga
     6961 tcactctttt ggagaattcg cttatttgct cgatgatctc atcaaggtag tgtttgtgtt
     7021 gttccacgaa cagctgcttc tgctcattat cttcgggaga ccctttgagc ttttcatagt
     7081 ggctggccag atacaagaaa ttaacgtatt tagagggcag tgccagctcg ttacctttct
     7141 gcagctcgcc cgcactagcg agcattcgtt tccggccgtt ttcaagctca aagagagagt
     7201 acttgggaag cttaatgatg aggtcttttt tgacctcttt atatcctttc gcctcgagaa
     7261 agtcgatggg gtttttttcg aagcttgatc gctccatgat tgtgatgccc agcagttcct
     7321 tgacgctttt gagtttttta gacttccctt tctccacttt ggccacaacc agtacactgt
     7381 aagcgactgt aggagaatcg aatccgccgt atttcttggg gtcccaatct tttttgcgtg
     7441 cgatcagctt gtcgctgttc cttttcggga ggatactttc cttggagaag cctccggtct
     7501 gtacttcggt ctttttaacg atgttcacct gcggcatgga caggaccttc cggactgtcg
     7561 cgaaatccct acccttgtcc cacacgattt ctcctgtttc tccgtttgtt tcgataagtg
     7621 gtcgcttccg aatctctcca ttggccagtg taatctcggt cttgaaaaaa ttcataatat
     7681 tgctgtaaaa gaagtactta gcggtggcct tgcctatttc ctgctcagac tttgcgatca
     7741 ttttcctaac atcgtacact ttatagtctc cgtaaacaaa ttcagattca agcttgggat
     7801 attttttgat aagtgcagtg cctaccactg cattcaggta ggcatcatgc gcatggtggt
     7861 aattgttgat ctctctcacc ttataaaact gaaagtcctt tctgaaatct gagaccagct
     7921 tagacttcag agtaataact ttcacctctc gaatcagttt gtcattttca tcgtacttgg
     7981 tgttcatgcg tgaatcgaga atttgggcca cgtgcttggt gatctggcgt gtctcaacaa
     8041 gctgcctttt gatgaagccg gctttatcca actcagacag gccacctcgt tcagccttag
     8101 tcagattatc gaacttccgt tgtgtgatca gtttggcgtt cagcagctgc cgccaataat
     8161 ttttcatttt cttgacaact tcttctgagg ggacgttatc actcttccct ctatttttat
     8221 cggatcttgt caacacttta ttatcaatag aatcatcttt gagaaaagac tggggcacga
     8281 tatgatccac gtcgtagtcg gagagccgat tgatgtccag ttcctgatcc acgtacatgt
     8341 ccctgccgtt ctgcaggtag tacaggtaga gcttctcatt ctgaagctgg gtgttttcaa
     8401 ctgggtgttc cttaaggatt tgggacccca gttcttttat accctcttca atcctcttca
     8461 tcctttccct actgttcttc tgtcccttct gggtagtttg gttctctcgg gccatctcga
     8521 taacgatatt ctcgggctta tgccttccca ttactttgac gagttcatcc acgaccttaa
     8581 cggtctgcag tattcccttt ttgatagctg ggctacctgc aagattagcg atgtgctcgt
     8641 gaagactgtc cccctggcca gaaacttgtg ctttctggat gtcctcctta aaggtgagag
     8701 agtcatcatg gatcaactgc atgaagttcc tgttggcaaa tccatcggac ttaagaaaat
     8761 ccaggattgt ctttccactc tgcttgtctc ggatcccatt gatcagtttt cttgacagcc
     8821 gcccccatcc tgtatatcgg cgcctcttga gctgtttcat gactttgtcg tcgaagagat
     8881 gagcgtaagt tttcaagcgt tcttcaatca tctccctatc ttcaaacaac gtaagggtga
     8941 ggacaatgtc ctcaagaatg tcctcgttct cctcattgtc caggaagtcc ttgtctttaa
     9001 tgattttcag gagatcgtga tacgttccca gggatgcgtt gaagcgatcc tccactccgc
     9061 tgatttcaac agagtcgaaa cattcaatct ttttgaaata gtcttctttg agctgtttca
     9121 cggtaacttt ccggttcgtc ttgaagagga ggtccacgat agctttcttc tgctctccag
     9181 acaggaatgc tggctttctc atcccttctg tgacgtattt gaccttggtg agctcgttat
     9241 aaactgtgaa gtactcgtac agcagagagt gtttaggaag caccttttcg ttaggcagat
     9301 ttttatcaaa gttagtcatc ctttcgatga aggactgggc agaggccccc ttatccacga
     9361 cttcctcgaa gttccaggga gtgatggtct cttctgattt gcgagtcatc cacgcgaatc
     9421 tggaatttcc ccgggcgagg gggcctacat agtagggtat ccgaaatgtg aggattttct
     9481 caatcttttc cctgttatct ttcaaaaagg ggtagaaatc ctcttgccgc ctgaggatag
     9541 cgtgcagttc gcccaggtga atctggtggg ggatgcttcc attgtcgaaa gtgcgctgtt
     9601 tgcgcaacag atcttctctg ttaagcttta ccagcagctc ctcggtgccg tccatttttt
     9661 ccaagatggg cttaataaat ttgtaaaatt cctcctggct tgctccgccg tcaatgtatc
     9721 cggcgtagcc atttttagac tgatcgaaga aaatttcctt gtacttctca ggcagttgct
     9781 gtctgacaag ggccttcagc aaagtcaagt cttggtggtg ctcatcatag cgcttgatca
     9841 tactagcgct cagcggagct ttggtgatct ccgtgttcac tcgcagaata tcactcagca
     9901 gaatggcgtc tgacaggttc tttgccgcca aaaaaaggtc tgcgtactgg tcgccgatct
     9961 gggccagcag attgtcgaga tcatcatcgt aggtgtcttt gctcagttga agcttggcat
    10021 cttcggccag gtcgaagtta gatttaaagt tgggggtcag cccgagtgac agggcgataa
    10081 gattaccaaa caggccgttc ttcttctccc cagggagctg tgcgatgagg ttttcgagcc
    10141 gccgggattt ggacagccta gcgctcagga ttgctttggc gtcaactccg gatgcgttga
    10201 tcgggttctc ttcgaaaagc tgattgtaag tctgaaccag ttggataaag agtttgtcga
    10261 catcgctgtt gtctgggttc aggtccccct cgatgaggaa gtgtccccga aatttgatca
    10321 tatgcgccag cgcgagatag atcaaccgca agtcagcctt atcagtactg tctacaagct
    10381 tcttcctcag atgatatatg gttgggtact tttcatggta cgccacctcg tccacgatat
    10441 tgccaaagat tgggtggcgc tcgtgctttt tatcctcctc caccaaaaag gactcctcca
    10501 gcctatggaa gaaagagtca tccaccttag ccatctcatt actaaagatc tcctgcaggt
    10561 agcagatccg attctttctg cgggtatatc tgcgccgtgc tgttcttttg agccgcgtgg
    10621 cttcggccgt ctccccggag tcgaacagga gggcgccaat gaggttcttc tttatgctgt
    10681 ggcgatcggt attgcccaga actttgaatt ttttgctcgg caccttgtac tcgtccgtaa
    10741 tgacggccca gccgacgctg tttgtgccga tatcgagccc aatggagtac ttcttgtcca
    10801 tgggtacccc gcggtgaatg cccaccttcc tcttcttctt gggggccatc ttgtcatcgt
    10861 catccttgta atcgatgtca tgatctttat aatcaccgtc atggtctttg tagtccatgg
    10921 tgctagctcc ggtccaacct gaaaaccgcg gagcctgctt ttttgtacaa acttgttgat
    10981 cggaattctc caggcgatct gacggttcac taaacgagct ctgcttatat aggcctccca
    11041 ccgtacacgc ctacctcgac atacgttctc tatcactgat agggagtaaa ctcgacatac
    11101 gttctctatc actgataggg ataaactcga catacgttct ctatcactga tagggagtaa
    11161 actcgacata cgttctctat cactgatagg gagtaaactc gacatacgtt ctctatcact
    11221 gatagggagt aaactcgaca tcgttctcta tcactgatag ggagtaaact cgacatacgt
    11281 tctctatcac tgatagggag taaactcgac agtactaagc ttgtcaccaa tcctgtccct
    11341 agtgggcggc cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt
    11401 gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
    11461 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
    11521 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
    11581 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
    11641 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
    11701 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
    11761 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
    11821 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
    11881 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
    11941 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
    12001 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
    12061 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
    12121 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
    12181 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
    12241 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
    12301 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
    12361 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
    12421 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
    12481 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
    12541 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
    12601 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
    12661 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
    12721 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
    12781 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
    12841 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
    12901 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
    12961 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
    13021 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
    13081 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
    13141 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
    13201 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
    13261 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
    13321 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
    13381 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
    13441 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
    13501 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
    13561 aaagtgc
//
