LOCUS       Exported               14433 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Tet-regulatable dCas9 lentiviral expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    pHAGE TRE dCas9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14433)
  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 14433)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14433
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    126..147
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        162..192
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    200..216
                     /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     205..227
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     224..240
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     224..240
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     290..309
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             501..1134
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    1181..1306
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1803..2036
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2221..2265
                     /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    2558..2675
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    2730..3000
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        3033..3071
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3068..3092
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3201..3225
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      3229..3238
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3235..7338
                     /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             7351..7371
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7372..7461
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     protein_bind    complement(7511..7535)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             7588..7614
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        7700..8911
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     8748..8767
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             8970..9716
                     /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    9874..10336
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(9930..9947)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     10157..10176
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             10352..11146
                     /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(10406..10425)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     11016..11035
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    11323..11911
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11376..11396)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11794..11805)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11986..12219
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(12338..12356)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        12424..12528
                     /gene="bla"
                     /label=AmpR promoter
     CDS             12529..13389
                     /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(12747..12766)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      13560..14148
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     14049..14068
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     14302..14319
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     14411..14430
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
       61 gcaaaaagct ccccgtggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
      121 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
      181 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
      241 atgattacga atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
      301 aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat
      361 catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta
      421 ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata
      481 tgtactacaa acttagtagt tggaagggct aattcactcc caaagaagac aagatatcct
      541 tgatctgtgg atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg
      601 gccaggggtc agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc
      661 agataaggta gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct
      721 gcatgggatg gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc
      781 atttcatcac gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc
      841 ttgctacaag ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg
      901 ggagtggcga gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc
      961 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     1021 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     1081 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     1141 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     1201 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     1261 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     1321 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     1381 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     1441 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     1501 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1561 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1621 aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat
     1681 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1741 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1801 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1861 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1921 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1981 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     2041 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     2101 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     2161 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     2221 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     2281 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     2341 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2401 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2461 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg
     2521 tatcgccgaa ttcacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg
     2581 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
     2641 aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag
     2701 atccagtttg gactaggatc ctttaccact ccctatcagt gatagagaaa agtgaaagtc
     2761 gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat
     2821 cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg
     2881 aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact
     2941 ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga
     3001 aaagtgaaag tcgagctcgg tacccgggtc gaggtaggcg tgtacggtgg gaggcctata
     3061 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
     3121 acctccatag aagacaccgg gaccgatcca gcctccgcgg ccccgaacta gtccagtgtg
     3181 gtggaattct gcagatatca acaagtttgt acaaaaaagc aggctggcgc caccatggac
     3241 aagaagtact ccattgggct cgctatcggt accaacagcg tcggctgggc cgtcattacg
     3301 gacgagtaca aggtgccgag caaaaaattc aaagttctgg gcaataccga tcgccacagc
     3361 ataaagaaga acctcattgg agccctcctg ttcgactccg gggagacggc cgaagccacg
     3421 cggctcaaaa gaacagcacg gcgcagatat acccgcagaa agaatcggat ctgctacctg
     3481 caggagatct ttagtaatga gatggctaag gtggatgact ctttcttcca taggctggag
     3541 gagtcctttt tggtggagga ggataaaaag cacgagcgcc acccaatctt tggcaatatc
     3601 gtggacgagg tggcgtacca tgaaaagtac ccaaccatat atcatctgag gaagaagctg
     3661 gtagacagta ctgataaggc tgacttgcgg ttgatctatc tcgcgctggc gcacatgatc
     3721 aaatttcggg gacacttcct catcgagggg gacctgaacc cagacaacag cgatgtcgac
     3781 aaactcttta tccaactggt tcagacttac aatcagcttt tcgaggagaa cccgatcaac
     3841 gcatccggcg ttgacgccaa agcaatcctg agcgctaggc tgtccaaatc ccggcggctc
     3901 gaaaacctca tcgcacagct ccctggggag aagaagaacg gcctgtttgg taatcttatc
     3961 gccctgtcac tcgggctgac ccccaacttt aaatctaact tcgacctggc cgaagatgcc
     4021 aagctgcaac tgagcaaaga cacctacgat gatgatctcg acaatctgct ggcccagatc
     4081 ggcgaccagt acgcagacct ttttttggcg gcaaagaacc tgtcagacgc cattctgctg
     4141 agtgatattc tgcgagtgaa cacggagatc accaaagctc cgctgagcgc tagtatgatc
     4201 aagcgctatg atgagcacca ccaagacttg actttgctga aggcccttgt cagacagcaa
     4261 ctgcctgaga agtacaagga aattttcttc gatcagtcta aaaatggcta cgccggatac
     4321 attgacggcg gagcaagcca ggaggaattt tacaaattta ttaagcccat cttggaaaaa
     4381 atggacggca ccgaggagct gctggtaaag ctgaacagag aagatctgtt gcgcaaacag
     4441 cgcactttcg acaatggaag catcccccac cagattcacc tgggcgaact gcacgctatc
     4501 ctcaggcggc aagaggattt ctaccccttt ttgaaagata acagggaaaa gattgagaaa
     4561 atcctcacat ttcggatacc ctactatgta ggccccctcg ctcggggaaa ttccagattc
     4621 gcgtggatga ctcgcaaatc agaagagacc atcactccct ggaacttcga ggaagtcgtg
     4681 gataaggggg cctctgccca gtccttcatc gaaaggatga ctaactttga taaaaatctg
     4741 cctaacgaaa aggtgcttcc taaacactct ctgctgtacg agtacttcac agtttataac
     4801 gagctcacca aggtcaaata cgtcacagaa gggatgagaa agccagcatt cctgtctgga
     4861 gagcagaaga aagctatcgt ggacctcctc ttcaagacga accggaaagt taccgtgaaa
     4921 cagctcaaag aagactattt caaaaagatt gaatgtttcg actctgttga aatcagcgga
     4981 gtggaggatc gcttcaacgc atccctggga acgtatcacg atctcctgaa aatcattaaa
     5041 gacaaggact tcctggacaa tgaggagaac gaggacattc ttgaggacat tgtcctcacc
     5101 cttacgttgt ttgaagatag ggagatgatt gaagaacgct tgaaaactta cgctcatctc
     5161 ttcgacgaca aagtcatgaa acagctcaag agacgccgat atacaggatg ggggcggctg
     5221 tcaagaaaac tgatcaatgg catccgagac aagcagagtg gaaagacaat cctggatttt
     5281 cttaagtccg atggatttgc caaccggaac ttcatgcagt tgatccatga tgactctctc
     5341 acctttaagg aggacatcca gaaagcacaa gtttctggcc agggggacag tcttcacgag
     5401 cacatcgcta atcttgcagg tagcccagct atcaaaaagg gaatactgca gaccgttaag
     5461 gtcgtggatg aactcgtcaa agtaatggga aggcataagc ccgagaatat cgttatcgag
     5521 atggcccgag agaaccaaac tacccagaag ggacagaaga acagtaggga aaggatgaag
     5581 aggattgaag agggtataaa agaactgggg tcccaaatcc ttaaggaaca cccagttgaa
     5641 aacacccagc ttcagaatga gaagctctac ctgtactacc tgcagaacgg cagggacatg
     5701 tacgtggatc aggaactgga catcaaccgg ttgtccgact acgacgtgga tgctatcgtg
     5761 ccccaaagct ttctcaaaga tgattctatt gataataaag tgttgacaag atccgataaa
     5821 aatagaggga agagtgataa cgtcccctca gaagaagttg tcaagaaaat gaaaaattat
     5881 tggcggcagc tgctgaacgc caaactgatc acacaacgga agttcgataa tctgactaag
     5941 gctgaacgag gtggcctgtc tgagttggat aaagccggct tcatcaaaag gcagcttgtt
     6001 gagacacgcc agatcaccaa gcacgtggcc caaattctcg attcacgcat gaacaccaag
     6061 tacgatgaaa atgacaaact gattcgagag gtgaaagtta ttactctgaa gtctaagctg
     6121 gtctcagatt tcagaaagga ctttcagttt tataaggtga gagagatcaa caattaccac
     6181 catgcgcatg atgcctacct gaatgcagtg gtaggcactg cacttatcaa aaaatatccc
     6241 aagctggaat ctgaatttgt ttacggagac tataaagtgt acgatgttag gaaaatgatc
     6301 gcaaagtctg agcaggaaat aggcaaggcc accgctaagt acttctttta cagcaatatt
     6361 atgaattttt tcaagaccga gattacactg gccaatggag agattcggaa gcgaccactt
     6421 atcgaaacaa acggagaaac aggagaaatc gtgtgggaca agggtaggga tttcgcgaca
     6481 gtccgcaagg tcctgtccat gccgcaggtg aacatcgtta aaaagaccga agtacagacc
     6541 ggaggcttct ccaaggaaag tatcctcccg aaaaggaaca gcgacaagct gatcgcacgc
     6601 aaaaaagatt gggaccccaa gaaatacggc ggattcgatt ctcctacagt cgcttacagt
     6661 gtactggttg tggccaaagt ggagaaaggg aagtctaaaa aactcaaaag cgtcaaggaa
     6721 ctgctgggca tcacaatcat ggagcgatcc agcttcgaga aaaaccccat cgactttctc
     6781 gaagcgaaag gatataaaga ggtcaaaaaa gacctcatca ttaagctgcc caagtactct
     6841 ctctttgagc ttgaaaacgg ccggaaacga atgctcgcta gtgcgggcga gctgcagaaa
     6901 ggtaacgagc tggcactgcc ctctaaatac gttaatttct tgtatctggc cagccactat
     6961 gaaaagctca aagggtctcc cgaagataat gagcagaagc agctgttcgt ggaacaacac
     7021 aaacactacc ttgatgagat catcgagcaa ataagcgagt tctccaaaag agtgatcctc
     7081 gccgacgcta acctcgataa ggtgctttct gcttacaata agcacaggga taagcccatc
     7141 agggagcagg cagaaaacat tatccacttg tttactctga ccaacttggg cgcgcctgca
     7201 gccttcaagt acttcgacac caccatagac agaaagcggt acacctctac aaaggaggtc
     7261 ctggacgcca cactgattca tcagtcaatt acggggctct atgaaacaag aatcgacctc
     7321 tctcagctcg gtggagacgg caccggcggg cccaagaaga agaggaaggt atacccatac
     7381 gatgttcctg actatgcggg ctatccctat gacgtcccgg actatgcagg atcgtatcct
     7441 tatgacgttc cagattacgc tggatcctaa tgattcgaat ccggataagc ggccgcctcg
     7501 agatatctag acccagcttt cttgtacaaa gtggttgata tccagcacag tggcggccgc
     7561 tcgagtctag agggcccgcg gttcgaatac ccatacgacg tcccagacta cgcttagtaa
     7621 tgattaatta aactagaaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg
     7681 agtaacccac ccaagatctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc
     7741 ctcctcacgg cgagcgctgc cacgtcagac gaagggcgca gcgagcgtcc tgatccttcc
     7801 gcccggacgc tcaggacagc ggcccgctgc tcataagact cggccttaga accccagtat
     7861 cagcagaagg acattttagg acgggacttg ggtgactcta gggcactggt tttctttcca
     7921 gagagcggaa caggcgagga aaagtagtcc cttctcggcg attctgcgga gggatctccg
     7981 tggggcggtg aacgccgatg attatataag gacgcgccgg gtgtggcaca gctagttccg
     8041 tcgcagccgg gatttgggtc gcggttcttg tttgtggatc gctgtgatcg tcacttggtg
     8101 agtagcgggc tgctgggctg gccggggctt tcgtggccgc cgggccgctc ggtgggacgg
     8161 aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc gcgagcaagg ttgccctgaa
     8221 ctgggggttg gggggagcgc agcaaaatgg cggctgttcc cgagtcttga atggaagacg
     8281 cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg gcggcaagaa
     8341 cccaaggtct tgaggccttc gctaatgcgg gaaagctctt attcgggtga gatgggctgg
     8401 ggcaccatct ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt
     8461 ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag
     8521 cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg
     8581 ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct
     8641 agggtaggct ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag
     8701 gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt
     8761 ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga
     8821 aatgtaatca tttgggtcaa tatgtaattt tcagtgttag actagtaaat tgtccgctaa
     8881 attctggccg tttttggctt ttttgttaga cgaagcttgg taccgagctc ggatctccac
     8941 cccgtaccgg tcctgcagtc gaattcacca tgtctagact ggacaagagc aaagtcataa
     9001 acggagctct ggaattactc aatggtgtcg gtatcgaagg cctgacgaca aggaaactcg
     9061 ctcaaaagct gggagttgag cagcctaccc tgtactggca cgtgaagaac aagcgggccc
     9121 tgctcgatgc cctgccaatc gagatgctgg acaggcatca tacccacttc tgccccctgg
     9181 aaggcgagtc atggcaagac tttctgcgga acaacgccaa gtcataccgc tgtgctctcc
     9241 tctcacatcg cgacggggct aaagtgcatc tcggcacccg cccaacagag aaacagtacg
     9301 aaaccctgga aaatcagctc gcgttcctgt gtcagcaagg cttctccctg gagaacgcac
     9361 tgtacgctct gtccgccgtg ggccacttta cactgggctg cgtattggag gaacaggagc
     9421 atcaagtagc aaaagaggaa agagagacac ctaccaccga ttctatgccc ccacttctga
     9481 gacaagcaat tgagctgttc gaccggcagg gagccgaacc tgccttcctt ttcggcctgg
     9541 aactaatcat atgtggcctg gagaaacagc taaagtgcga aagcggcggg ccgaccgacg
     9601 cccttgacga ttttgactta gacatgctcc cagccgatgc ccttgacgac tttgaccttg
     9661 atatgctgcc tgctgacgct cttgacgatt ttgaccttga catgctcccc gggtaactaa
     9721 gtaaggatcc gcggccgcac tagaggaatt ccgcccctct cccccccccc cctccccccc
     9781 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     9841 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     9901 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     9961 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
    10021 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
    10081 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
    10141 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
    10201 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
    10261 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
    10321 aaaaacacga tgataagctt accggtccac catgattgaa caagatggat tgcacgcagg
    10381 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
    10441 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
    10501 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
    10561 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
    10621 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
    10681 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
    10741 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
    10801 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
    10861 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
    10921 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
    10981 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
    11041 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
    11101 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgatgta caagtaaagc
    11161 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
    11221 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgttt
    11281 agtccctccc aattcgatat caagcttatc gcgatagatc ctaatcaacc tctggattac
    11341 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
    11401 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
    11461 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
    11521 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
    11581 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
    11641 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
    11701 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
    11761 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
    11821 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
    11881 agtcggatct ccctttgggc cgcctccccg cctgagatcc tttaagacca atgacttaca
    11941 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
    12001 actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga
    12061 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
    12121 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
    12181 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta
    12241 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg cccgggttaa
    12301 ttaaggaaag ggctagatca ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
    12361 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
    12421 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
    12481 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
    12541 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
    12601 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
    12661 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
    12721 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
    12781 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
    12841 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
    12901 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
    12961 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
    13021 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
    13081 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
    13141 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
    13201 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
    13261 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
    13321 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
    13381 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
    13441 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
    13501 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
    13561 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
    13621 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
    13681 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
    13741 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
    13801 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
    13861 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
    13921 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
    13981 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
    14041 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
    14101 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
    14161 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
    14221 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
    14281 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
    14341 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagcaagct catggctgac
    14401 taattttttt tatttatgca gaggccgagg ccg
//
