pLV-dCas9-p300-P2A-PuroR

LOCUS       pLV-dCas9-p300-P2A-PuroR              14538 bp ds-DNA     circular SYN 18-MAY-2020
DEFINITION  Lentiviral Sp dCas9-p300-P2A-PuroR.
ACCESSION   .
VERSION     .
KEYWORDS    pLV-dCas9-p300-P2A-PuroR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14538)
  AUTHORS   Klann TS, Black JB, Chellappan M, Safi A, Song L, Hilton IB, 
            Crawford GE, Reddy TE, Gersbach CA
  TITLE     CRISPR-Cas9 epigenome editing enables high-throughput screening for 
            functional regulatory elements in the human genome.
  JOURNAL   Nat Biotechnol. 2017 Apr 3. doi: 10.1038/nbt.3853.
  PUBMED    28369033
REFERENCE   2  (bases 1 to 14538)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 18, 2020 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14538
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2440..2651
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2670..2679
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2676..6779
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6780..6827
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6834..8684
                     /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      8448..8457
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             8691..8714
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8724..8780
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             8781..9377
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     9274..9294
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9393..9981
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9446..9466)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9864..9875)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10053..10286
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10312..10329)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10318..10542
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10588..11016
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10675..10694)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10885..10906
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11025..11045)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11030..11359
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11210..11345
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11272..11291
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11407..11454
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11473..11847
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    11977..12098
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12014..12033)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12068..12087
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12147..12163)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12147..12163)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12160..12182)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12171..12187
                     /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(12195..12225)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12240..12261
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12378..12395)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12549..13137)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12629..12648)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(13308..14168)
                     /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     13931..13950
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14169..14273)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14348..14367)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttaaggtacc gaggcgatag ggttaaggga gctagctagg
     2401 tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca
     2461 cagtccccga gaagttgggg ggaggggtcg gcaattgatc cggtgcctag agaaggtggc
     2521 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
     2581 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
     2641 ccagaacaca ggaccggttc tagagcgctg ccaccatgga caagaagtac agcatcggcc
     2701 tggccatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
     2761 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
     2821 gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca
     2881 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
     2941 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
     3001 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
     3061 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
     3121 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
     3181 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
     3241 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
     3301 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc
     3361 tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga
     3421 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg
     3481 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
     3541 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
     3601 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc
     3661 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
     3721 agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc
     3781 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac
     3841 tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca
     3901 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt
     3961 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc
     4021 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga
     4081 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc
     4141 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc
     4201 ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat
     4261 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg
     4321 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
     4381 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
     4441 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
     4501 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
     4561 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
     4621 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
     4681 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
     4741 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
     4801 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
     4861 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
     4921 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
     4981 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
     5041 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
     5101 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
     5161 acatcaaccg gctgtccgac tacgatgtgg acgccatcgt gcctcagagc tttctgaagg
     5221 acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca
     5281 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg
     5341 ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga
     5401 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
     5461 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc
     5521 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
     5581 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
     5641 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg
     5701 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
     5761 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
     5821 agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa
     5881 ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca
     5941 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt
     6001 ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta
     6061 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag
     6121 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca
     6181 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag
     6241 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg
     6301 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc
     6361 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc
     6421 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga
     6481 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca
     6541 aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata
     6601 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca
     6661 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc
     6721 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca
     6781 agcgacctgc cgccacaaag aaggctggac aggctaagaa gaagaaagga tccattttca
     6841 aaccagaaga actacgacag gcactgatgc caactttgga ggcactttac cgtcaggatc
     6901 cagaatccct tccctttcgt caacctgtgg accctcagct tttaggaatc cctgattact
     6961 ttgatattgt gaagagcccc atggatcttt ctaccattaa gaggaagtta gacactggac
     7021 agtatcagga gccctggcag tatgtcgatg atatttggct tatgttcaat aatgcctggt
     7081 tatataaccg gaaaacatca cgggtataca aatactgctc caagctctct gaggtctttg
     7141 aacaagaaat tgacccagtg atgcaaagcc ttggatactg ttgtggcaga aagttggagt
     7201 tctctccaca gacactgtgt tgctacggca aacagttgtg cacaatacct cgtgatgcca
     7261 cttattacag ttaccagaac aggtatcatt tctgtgagaa gtgtttcaat gagatccaag
     7321 gggagagcgt ttctttgggg gatgaccctt cccagcctca aactacaata aataaagaac
     7381 aattttccaa gagaaaaaat gacacactgg atcctgaact gtttgttgaa tgtacagagt
     7441 gcggaagaaa gatgcatcag atctgtgtcc ttcaccatga gatcatctgg cctgctggat
     7501 tcgtctgtga tggctgttta aagaaaagtg cacgaactag gaaagaaaat aagttttctg
     7561 ctaaaaggtt gccatctacc agacttggca cctttctaga gaatcgtgtg aatgactttc
     7621 tgaggcgaca gaatcaccct gagtcaggag aggtcactgt tagagtagtt catgcttctg
     7681 acaaaaccgt ggaagtaaaa ccaggcatga aagcaaggtt tgtggacagt ggagagatgg
     7741 cagaatcctt tccataccga accaaagccc tctttgcctt tgaagaaatt gatggtgttg
     7801 acctgtgctt ctttggcatg catgttcaag agtatggctc tgactgccct ccacccaacc
     7861 agaggagagt atacatatct tacctcgata gtgttcattt cttccgtcct aaatgcttga
     7921 ggactgcagt ctatcatgaa atcctaattg gatatttaga atatgtcaag aaattaggtt
     7981 acacaacagg gcatatttgg gcatgtccac caagtgaggg agatgattat atcttccatt
     8041 gccatcctcc tgaccagaag atacccaagc ccaagcgact gcaggaatgg tacaaaaaaa
     8101 tgcttgacaa ggctgtatca gagcgtattg tccatgacta caaggatatt tttaaacaag
     8161 ctactgaaga tagattaaca agtgcaaagg aattgcctta tttcgagggt gatttctggc
     8221 ccaatgttct ggaagaaagc attaaggaac tggaacagga ggaagaagag agaaaacgag
     8281 aggaaaacac cagcaatgaa agcacagatg tgaccaaggg agacagcaaa aatgctaaaa
     8341 agaagaataa taagaaaacc agcaaaaata agagcagcct gagtaggggc aacaagaaga
     8401 aacccgggat gcccaatgta tctaacgacc tctcacagaa actatatgcc accatggaga
     8461 agcataaaga ggtcttcttt gtgatccgcc tcattgctgg ccctgctgcc aactccctgc
     8521 ctcccattgt tgatcctgat cctctcatcc cctgcgatct gatggatggt cgggatgcgt
     8581 ttctcacgct ggcaagggac aagcacctgg agttctcttc actccgaaga gcccagtggt
     8641 ccaccatgtg catgctggtg gagctgcaca cgcagagcca ggacggatcc gattacaaag
     8701 acgatgacga taagggatcc ggcgcaacaa acttctctct gctgaaacaa gccggagatg
     8761 tcgaagagaa tcctggaccg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     8821 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     8881 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     8941 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     9001 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     9061 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     9121 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggagtctcg cccgaccacc
     9181 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     9241 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     9301 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     9361 gcaagcccgg tgcctgaacg cgttaagtcg acaatcaacc tctggattac aaaatttgtg
     9421 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     9481 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     9541 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     9601 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     9661 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     9721 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     9781 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     9841 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     9901 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     9961 ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga
    10021 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
    10081 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
    10141 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
    10201 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
    10261 ttagtcagtg tggaaaatct ctagcagggc ccgtttaaac ccgctgatca gcctcgactg
    10321 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
    10381 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
    10441 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
    10501 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
    10561 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
    10621 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
    10681 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
    10741 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
    10801 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
    10861 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
    10921 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
    10981 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
    11041 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
    11101 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
    11161 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
    11221 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
    11281 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
    11341 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
    11401 gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
    11461 aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
    11521 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
    11581 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
    11641 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
    11701 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
    11761 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
    11821 cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc
    11881 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
    11941 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
    12001 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
    12061 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
    12121 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
    12181 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
    12241 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
    12301 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
    12361 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
    12421 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
    12481 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
    12541 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
    12601 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
    12661 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
    12721 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
    12781 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
    12841 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
    12901 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
    12961 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
    13021 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
    13081 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
    13141 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
    13201 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
    13261 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
    13321 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
    13381 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
    13441 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
    13501 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
    13561 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
    13621 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
    13681 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
    13741 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
    13801 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
    13861 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
    13921 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
    13981 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
    14041 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
    14101 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
    14161 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
    14221 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
    14281 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
    14341 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
    14401 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
    14461 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
    14521 gggccagata tacgcgtt
//