LOCUS       Exported               14606 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Tet-regulatable dCas9-VP64 2nd generation lentiviral expression 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    pHAGE TRE dCas9-VP64
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14606)
  AUTHORS   Kearns NA, Genga RM, Enuameh MS, Garber M, Wolfe SA, Maehr R
  TITLE     Cas9 effector-mediated regulation of transcription and 
            differentiation in human pluripotent stem cells.
  JOURNAL   Development. 2014 Jan;141(1):219-23. doi: 10.1242/dev.103341.
  PUBMED    24346702
REFERENCE   2  (bases 1 to 14606)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14606
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             10..54
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    347..464
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    519..789
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        822..860
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     857..881
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    990..1014
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1018..1027
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1024..5127
                     /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             5140..5160
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5161..5250
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             5272..5421
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     protein_bind    complement(5468..5492)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5545..5571
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        5657..6868
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     6705..6724
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             6927..7673
                     /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
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    7725..8298
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7892..7909)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8119..8138
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             8314..9108
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8368..8387)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8978..8997
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    9285..9873
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9338..9358)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9756..9767)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9948..10181
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10300..10318)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        10386..10490
                     /gene="bla"
                     /label=AmpR promoter
     CDS             10491..11351
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(10709..10728)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11522..12110
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12011..12030
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12264..12281
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     12373..12392
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     protein_bind    12521..12542
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        12557..12587
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12595..12611
                     /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)."
     primer_bind     12600..12622
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12619..12635
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12619..12635
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12685..12704
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             12896..13529
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    13576..13701
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    14198..14431
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
       61 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
      121 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
      181 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa
      241 ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga
      301 tctcgacggt atcgccgaat tcacaaatgg cagtattcat ccacaatttt aaaagaaaag
      361 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
      421 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
      481 acagcagaga tccagtttgg actaggatcc tttaccactc cctatcagtg atagagaaaa
      541 gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
      601 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
      661 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
      721 tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag
      781 tgatagagaa aagtgaaagt cgagctcggt acccgggtcg aggtaggcgt gtacggtggg
      841 aggcctatat aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac
      901 gctgttttga cctccataga agacaccggg accgatccag cctccgcggc cccgaactag
      961 tccagtgtgg tggaattctg cagatatcaa caagtttgta caaaaaagca ggctggcgcc
     1021 accatggaca agaagtactc cattgggctc gctatcggta ccaacagcgt cggctgggcc
     1081 gtcattacgg acgagtacaa ggtgccgagc aaaaaattca aagttctggg caataccgat
     1141 cgccacagca taaagaagaa cctcattgga gccctcctgt tcgactccgg ggagacggcc
     1201 gaagccacgc ggctcaaaag aacagcacgg cgcagatata cccgcagaaa gaatcggatc
     1261 tgctacctgc aggagatctt tagtaatgag atggctaagg tggatgactc tttcttccat
     1321 aggctggagg agtccttttt ggtggaggag gataaaaagc acgagcgcca cccaatcttt
     1381 ggcaatatcg tggacgaggt ggcgtaccat gaaaagtacc caaccatata tcatctgagg
     1441 aagaagctgg tagacagtac tgataaggct gacttgcggt tgatctatct cgcgctggcg
     1501 cacatgatca aatttcgggg acacttcctc atcgaggggg acctgaaccc agacaacagc
     1561 gatgtcgaca aactctttat ccaactggtt cagacttaca atcagctttt cgaggagaac
     1621 ccgatcaacg catccggcgt tgacgccaaa gcaatcctga gcgctaggct gtccaaatcc
     1681 cggcggctcg aaaacctcat cgcacagctc cctggggaga agaagaacgg cctgtttggt
     1741 aatcttatcg ccctgtcact cgggctgacc cccaacttta aatctaactt cgacctggcc
     1801 gaagatgcca agctgcaact gagcaaagac acctacgatg atgatctcga caatctgctg
     1861 gcccagatcg gcgaccagta cgcagacctt tttttggcgg caaagaacct gtcagacgcc
     1921 attctgctga gtgatattct gcgagtgaac acggagatca ccaaagctcc gctgagcgct
     1981 agtatgatca agcgctatga tgagcaccac caagacttga ctttgctgaa ggcccttgtc
     2041 agacagcaac tgcctgagaa gtacaaggaa attttcttcg atcagtctaa aaatggctac
     2101 gccggataca ttgacggcgg agcaagccag gaggaatttt acaaatttat taagcccatc
     2161 ttggaaaaaa tggacggcac cgaggagctg ctggtaaagc tgaacagaga agatctgttg
     2221 cgcaaacagc gcactttcga caatggaagc atcccccacc agattcacct gggcgaactg
     2281 cacgctatcc tcaggcggca agaggatttc tacccctttt tgaaagataa cagggaaaag
     2341 attgagaaaa tcctcacatt tcggataccc tactatgtag gccccctcgc tcggggaaat
     2401 tccagattcg cgtggatgac tcgcaaatca gaagagacca tcactccctg gaacttcgag
     2461 gaagtcgtgg ataagggggc ctctgcccag tccttcatcg aaaggatgac taactttgat
     2521 aaaaatctgc ctaacgaaaa ggtgcttcct aaacactctc tgctgtacga gtacttcaca
     2581 gtttataacg agctcaccaa ggtcaaatac gtcacagaag ggatgagaaa gccagcattc
     2641 ctgtctggag agcagaagaa agctatcgtg gacctcctct tcaagacgaa ccggaaagtt
     2701 accgtgaaac agctcaaaga agactatttc aaaaagattg aatgtttcga ctctgttgaa
     2761 atcagcggag tggaggatcg cttcaacgca tccctgggaa cgtatcacga tctcctgaaa
     2821 atcattaaag acaaggactt cctggacaat gaggagaacg aggacattct tgaggacatt
     2881 gtcctcaccc ttacgttgtt tgaagatagg gagatgattg aagaacgctt gaaaacttac
     2941 gctcatctct tcgacgacaa agtcatgaaa cagctcaaga gacgccgata tacaggatgg
     3001 gggcggctgt caagaaaact gatcaatggc atccgagaca agcagagtgg aaagacaatc
     3061 ctggattttc ttaagtccga tggatttgcc aaccggaact tcatgcagtt gatccatgat
     3121 gactctctca cctttaagga ggacatccag aaagcacaag tttctggcca gggggacagt
     3181 cttcacgagc acatcgctaa tcttgcaggt agcccagcta tcaaaaaggg aatactgcag
     3241 accgttaagg tcgtggatga actcgtcaaa gtaatgggaa ggcataagcc cgagaatatc
     3301 gttatcgaga tggcccgaga gaaccaaact acccagaagg gacagaagaa cagtagggaa
     3361 aggatgaaga ggattgaaga gggtataaaa gaactggggt cccaaatcct taaggaacac
     3421 ccagttgaaa acacccagct tcagaatgag aagctctacc tgtactacct gcagaacggc
     3481 agggacatgt acgtggatca ggaactggac atcaaccggt tgtccgacta cgacgtggat
     3541 gctatcgtgc cccaaagctt tctcaaagat gattctattg ataataaagt gttgacaaga
     3601 tccgataaaa atagagggaa gagtgataac gtcccctcag aagaagttgt caagaaaatg
     3661 aaaaattatt ggcggcagct gctgaacgcc aaactgatca cacaacggaa gttcgataat
     3721 ctgactaagg ctgaacgagg tggcctgtct gagttggata aagccggctt catcaaaagg
     3781 cagcttgttg agacacgcca gatcaccaag cacgtggccc aaattctcga ttcacgcatg
     3841 aacaccaagt acgatgaaaa tgacaaactg attcgagagg tgaaagttat tactctgaag
     3901 tctaagctgg tctcagattt cagaaaggac tttcagtttt ataaggtgag agagatcaac
     3961 aattaccacc atgcgcatga tgcctacctg aatgcagtgg taggcactgc acttatcaaa
     4021 aaatatccca agctggaatc tgaatttgtt tacggagact ataaagtgta cgatgttagg
     4081 aaaatgatcg caaagtctga gcaggaaata ggcaaggcca ccgctaagta cttcttttac
     4141 agcaatatta tgaatttttt caagaccgag attacactgg ccaatggaga gattcggaag
     4201 cgaccactta tcgaaacaaa cggagaaaca ggagaaatcg tgtgggacaa gggtagggat
     4261 ttcgcgacag tccgcaaggt cctgtccatg ccgcaggtga acatcgttaa aaagaccgaa
     4321 gtacagaccg gaggcttctc caaggaaagt atcctcccga aaaggaacag cgacaagctg
     4381 atcgcacgca aaaaagattg ggaccccaag aaatacggcg gattcgattc tcctacagtc
     4441 gcttacagtg tactggttgt ggccaaagtg gagaaaggga agtctaaaaa actcaaaagc
     4501 gtcaaggaac tgctgggcat cacaatcatg gagcgatcca gcttcgagaa aaaccccatc
     4561 gactttctcg aagcgaaagg atataaagag gtcaaaaaag acctcatcat taagctgccc
     4621 aagtactctc tctttgagct tgaaaacggc cggaaacgaa tgctcgctag tgcgggcgag
     4681 ctgcagaaag gtaacgagct ggcactgccc tctaaatacg ttaatttctt gtatctggcc
     4741 agccactatg aaaagctcaa agggtctccc gaagataatg agcagaagca gctgttcgtg
     4801 gaacaacaca aacactacct tgatgagatc atcgagcaaa taagcgagtt ctccaaaaga
     4861 gtgatcctcg ccgacgctaa cctcgataag gtgctttctg cttacaataa gcacagggat
     4921 aagcccatca gggagcaggc agaaaacatt atccacttgt ttactctgac caacttgggc
     4981 gcgcctgcag ccttcaagta cttcgacacc accatagaca gaaagcggta cacctctaca
     5041 aaggaggtcc tggacgccac actgattcat cagtcaatta cggggctcta tgaaacaaga
     5101 atcgacctct ctcagctcgg tggagacggc accggcgggc ccaagaagaa gaggaaggta
     5161 tacccatacg atgttcctga ctatgcgggc tatccctatg acgtcccgga ctatgcagga
     5221 tcgtatcctt atgacgttcc agattacgct ggatccggtt ccggacgggc tgacgcattg
     5281 gacgattttg atctggatat gctgggaagt gacgccctcg atgattttga ccttgacatg
     5341 cttggttcgg atgcccttga tgactttgac ctcgacatgc tcggcagtga cgcccttgat
     5401 gatttcgacc tggacatgct gattaactaa ttcgaatccg gataagcggc cgcctcgaga
     5461 tatctagacc cagctttctt gtacaaagtg gttgatatcc agcacagtgg cggccgctcg
     5521 agtctagagg gcccgcggtt cgaataccca tacgacgtcc cagactacgc ttagtaatga
     5581 ttaattaaac tagaaattct accgggtagg ggaggcgctt ttcccaaggc agtctggagt
     5641 aacccaccca agatctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc
     5701 ctcacggcga gcgctgccac gtcagacgaa gggcgcagcg agcgtcctga tccttccgcc
     5761 cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag
     5821 cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag
     5881 agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg
     5941 ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg
     6001 cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt
     6061 agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt gggacggaag
     6121 cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg
     6181 ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt
     6241 gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc
     6301 aaggtcttga ggccttcgct aatgcgggaa agctcttatt cgggtgagat gggctggggc
     6361 accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg
     6421 ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc
     6481 gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca
     6541 cctgccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg
     6601 gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg
     6661 tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg
     6721 aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat
     6781 gtaatcattt gggtcaatat gtaattttca gtgttagact agtaaattgt ccgctaaatt
     6841 ctggccgttt ttggcttttt tgttagacga agcttggtac cgagctcgga tctccacccc
     6901 gtaccggtcc tgcagtcgaa ttcaccatgt ctagactgga caagagcaaa gtcataaacg
     6961 gagctctgga attactcaat ggtgtcggta tcgaaggcct gacgacaagg aaactcgctc
     7021 aaaagctggg agttgagcag cctaccctgt actggcacgt gaagaacaag cgggccctgc
     7081 tcgatgccct gccaatcgag atgctggaca ggcatcatac ccacttctgc cccctggaag
     7141 gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc ataccgctgt gctctcctct
     7201 cacatcgcga cggggctaaa gtgcatctcg gcacccgccc aacagagaaa cagtacgaaa
     7261 ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt ctccctggag aacgcactgt
     7321 acgctctgtc cgccgtgggc cactttacac tgggctgcgt attggaggaa caggagcatc
     7381 aagtagcaaa agaggaaaga gagacaccta ccaccgattc tatgccccca cttctgagac
     7441 aagcaattga gctgttcgac cggcagggag ccgaacctgc cttccttttc ggcctggaac
     7501 taatcatatg tggcctggag aaacagctaa agtgcgaaag cggcgggccg accgacgccc
     7561 ttgacgattt tgacttagac atgctcccag ccgatgccct tgacgacttt gaccttgata
     7621 tgctgcctgc tgacgctctt gacgattttg accttgacat gctccccggg taactaagta
     7681 aggatccgcg gccgcactag aggaattccg cccctctccc cccccccctc tccctccccc
     7741 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     7801 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     7861 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
     7921 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     7981 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     8041 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     8101 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     8161 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt
     8221 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     8281 tgaaaaacac gatgataagc ttaccggtcc accatgattg aacaagatgg attgcacgca
     8341 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     8401 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     8461 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     8521 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     8581 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     8641 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     8701 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     8761 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     8821 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     8881 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     8941 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     9001 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     9061 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgatg tacaagtaaa
     9121 gcggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     9181 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     9241 ttagtccctc ccaattcgat atcaagctta tcgcgataga tcctaatcaa cctctggatt
     9301 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     9361 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     9421 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     9481 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     9541 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     9601 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     9661 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     9721 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     9781 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     9841 cgagtcggat ctccctttgg gccgcctccc cgcctgagat cctttaagac caatgactta
     9901 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     9961 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca
    10021 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
    10081 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
    10141 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
    10201 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggcccgggtt
    10261 aattaaggaa agggctagat cattcttgaa gacgaaaggg cctcgtgata cgcctatttt
    10321 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
    10381 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
    10441 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
    10501 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
    10561 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
    10621 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
    10681 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
    10741 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
    10801 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
    10861 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
    10921 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
    10981 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
    11041 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
    11101 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
    11161 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
    11221 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
    11281 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
    11341 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
    11401 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
    11461 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
    11521 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
    11581 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
    11641 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
    11701 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
    11761 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
    11821 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
    11881 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
    11941 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
    12001 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
    12061 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
    12121 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
    12181 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
    12241 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
    12301 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagcaag ctcatggctg
    12361 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
    12421 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctccccg tggcacgaca
    12481 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
    12541 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
    12601 gcggataaca atttcacaca ggaaacagct atgacatgat tacgaatttc acaaataaag
    12661 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
    12721 tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc
    12781 ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat tcctctgaat
    12841 tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagttggaa
    12901 gggctaattc actcccaaag aagacaagat atccttgatc tgtggatcta ccacacacaa
    12961 ggctacttcc ctgattagca gaactacaca ccagggccag gggtcagata tccactgacc
    13021 tttggatggt gctacaagct agtaccagtt gagccagata aggtagaaga ggccaataaa
    13081 ggagagaaca ccagcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga
    13141 gaagtgttag agtggaggtt tgacagccgc ctagcatttc atcacgtggc ccgagagctg
    13201 catccggagt acttcaagaa ctgctgatat cgagcttgct acaagggact ttccgctggg
    13261 gactttccag ggaggcgtgg cctgggcggg actggggagt ggcgagccct cagatcctgc
    13321 atataagcag ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg
    13381 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
    13441 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
    13501 cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa
    13561 gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga
    13621 ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag
    13681 agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa
    13741 attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc
    13801 agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga
    13861 caaatactgg gacagctaca accatccctt cagacaggat cagaagaact tagatcatta
    13921 tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag
    13981 gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg
    14041 gccggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat
    14101 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
    14161 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
    14221 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac
    14281 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
    14341 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
    14401 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
    14461 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt
    14521 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
    14581 actccttaat tgaagaatcg caaaac
//
