LOCUS       sequence-313417-       11402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminal Flag and biotin-acceptor-site (FB)-tagged dCas9
            protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11402)
  AUTHORS   Liu X, Zhang Y, Chen Y, Li M, Zhou F, Li K, Cao H, Ni M, Liu Y, Gu 
            Z, Dickerson KE, Xie S, Hon GC, Xuan Z, Zhang MQ, Shao Z, Xu J
  TITLE     In Situ Capture of Chromatin Interactions by Biotinylated dCas9.
  JOURNAL   Cell. 2017 Aug 24;170(5):1028-1043.e19. doi: 
            10.1016/j.cell.2017.08.003.
  PUBMED    28841410
REFERENCE   2  (bases 1 to 11402)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2017.08"; journalName: "Cell"; date: "2017-08-24- 
            24"; volume: "170"; issue: "5"; pages: "1028-1043.e19"
COMMENT     Sequence Label: pEF1a-FB-dCas9-puro
FEATURES             Location/Qualifiers
     source          1..11402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1212..1235
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1275..1319
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             1350..1370
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1371..1394,1395..1397,1398..3884,3885..3887,
                     3888..5471)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5481..5501
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6405..6421)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6503..6519)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6504..6520)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(6548..6566)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6587..6603)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6611..6627
                     /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(join(6635..6641,6642..6659,6660..6665))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6680..6701
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6818..6835)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6989..7577)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7069..7088)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7748..8539,8540..8608))
                     /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     8371..8390
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8609..8713)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8740..9195
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8827..8846)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9037..9058
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9336..9352
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9359..9378
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9359..9377
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    9472..9696
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     9685..9702
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(9777..10376)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(9860..9880)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     10357..10376
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(10388..10887)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(10436..10455)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     10829..10852
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    10952..10973
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10988..10993,10994..11011,11012..11018)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11026..11042
                     /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     11050..11066
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        11098..11308
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        11120..11316
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      11167..11302
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattgatcc agatctttcg
     1201 aaatcaccat ggactacaaa gacgatgacg ataaagcaag aagagcatct gtggaattcc
     1261 tgcagcccgg gtccggcctg aacgacatct tcgaggctca gaaaatcgaa tggcacgaag
     1321 gcgcgccgag ctcgaggatc cttgctagcc ccaaaaagaa gaggaaagtg gacaagaagt
     1381 attctatcgg actggccatc gggactaata gcgtcgggtg ggccgtgatc actgacgagt
     1441 acaaggtgcc ctctaagaag ttcaaggtgc tcgggaacac cgaccggcat tccatcaaga
     1501 aaaatctgat cggagctctc ctctttgatt caggggagac cgctgaagca acccgcctca
     1561 agcggactgc tagacggcgg tacaccagga ggaagaaccg gatttgttac cttcaagaga
     1621 tattctccaa cgaaatggca aaggtcgacg acagcttctt ccataggctg gaagaatcat
     1681 tcctcgtgga agaggataag aagcatgaac ggcatcccat cttcggtaat atcgtcgacg
     1741 aggtggccta tcacgagaaa tacccaacca tctaccatct tcgcaaaaag ctggtggact
     1801 caaccgacaa ggcagacctc cggcttatct acctggccct ggcccacatg atcaagttca
     1861 gaggccactt cctgatcgag ggcgacctca atcctgacaa tagcgatgtg gataaactgt
     1921 tcatccagct ggtgcagact tacaaccagc tctttgaaga gaaccccatc aatgcaagcg
     1981 gagtcgatgc caaggccatt ctgtcagccc ggctgtcaaa gagccgcaga cttgagaatc
     2041 ttatcgctca gctgccgggt gaaaagaaaa atggactgtt cgggaacctg attgctcttt
     2101 cacttgggct gactcccaat ttcaagtcta atttcgacct ggcagaggat gccaagctgc
     2161 aactgtccaa ggacacctat gatgacgatc tcgacaacct cctggcccag atcggtgacc
     2221 aatacgccga ccttttcctt gctgctaaga atctttctga cgccatcctg ctgtctgaca
     2281 ttctccgcgt gaacactgaa atcaccaagg cccctctttc agcttcaatg attaagcggt
     2341 atgatgagca ccaccaggac ctgaccctgc ttaaggcact cgtccggcag cagcttccgg
     2401 agaagtacaa ggaaatcttc tttgaccagt caaagaatgg atacgccggc tacatcgacg
     2461 gaggtgcctc ccaagaggaa ttttataagt ttatcaaacc tatccttgag aagatggacg
     2521 gcaccgaaga gctcctcgtg aaactgaatc gggaggatct gctgcggaag cagcgcactt
     2581 tcgacaatgg gagcattccc caccagatcc atcttgggga gcttcacgcc atccttcggc
     2641 gccaagagga cttctacccc tttcttaagg acaacaggga gaagattgag aaaattctca
     2701 ctttccgcat cccctactac gtgggacccc tcgccagagg aaatagccgg tttgcttgga
     2761 tgaccagaaa gtcagaagaa actatcactc cctggaactt cgaagaggtg gtggacaagg
     2821 gagccagcgc tcagtcattc atcgaacgga tgactaactt cgataagaac ctccccaatg
     2881 agaaggtcct gccgaaacat tccctgctct acgagtactt taccgtgtac aacgagctga
     2941 ccaaggtgaa atatgtcacc gaagggatga ggaagcccgc attcctgtca ggcgaacaaa
     3001 agaaggcaat tgtggacctt ctgttcaaga ccaatagaaa ggtgaccgtg aagcagctga
     3061 aggaggacta tttcaagaaa attgaatgct tcgactctgt ggagattagc ggggtcgaag
     3121 atcggttcaa cgcaagcctg ggtacctacc atgatctgct taagatcatc aaggacaagg
     3181 attttctgga caatgaggag aacgaggaca tccttgagga cattgtcctg actctcactc
     3241 tgttcgagga ccgggaaatg atcgaggaga ggcttaagac ctacgcccat ctgttcgacg
     3301 ataaagtgat gaagcaactt aaacggagaa gatataccgg atggggacgc cttagccgca
     3361 aactcatcaa cggaatccgg gacaaacaga gcggaaagac cattcttgat ttccttaaga
     3421 gcgacggatt cgctaatcgc aacttcatgc aacttatcca tgatgattcc ctgaccttta
     3481 aggaggacat ccagaaggcc caagtgtctg gacaaggtga ctcactgcac gagcatatcg
     3541 caaatctggc tggttcaccc gctattaaga agggtattct ccagaccgtg aaagtcgtgg
     3601 acgagctggt caaggtgatg ggtcgccata aaccagagaa cattgtcatc gagatggcca
     3661 gggaaaacca gactacccag aagggacaga agaacagcag ggagcggatg aaaagaattg
     3721 aggaagggat taaggagctc gggtcacaga tccttaaaga gcacccggtg gaaaacaccc
     3781 agcttcagaa tgagaagctc tatctgtact accttcaaaa tggacgcgat atgtatgtgg
     3841 accaagagct tgatatcaac aggctctcag actacgacgt ggacgccatc gtccctcaga
     3901 gcttcctcaa agacgactca attgacaata aggtgctgac tcgctcagac aagaaccggg
     3961 gaaagtcaga taacgtgccc tcagaggaag tcgtgaaaaa gatgaagaac tattggcgcc
     4021 agcttctgaa cgcaaagctg atcactcagc ggaagttcga caatctcact aaggctgaga
     4081 ggggcggact gagcgaactg gacaaagcag gattcattaa acggcaactt gtggagactc
     4141 ggcagattac taaacatgtc gcccaaatcc ttgactcacg catgaatacc aagtacgacg
     4201 aaaacgacaa acttatccgc gaggtgaagg tgattaccct gaagtccaag ctggtcagcg
     4261 atttcagaaa ggactttcaa ttctacaaag tgcgggagat caataactat catcatgctc
     4321 atgacgcata tctgaatgcc gtggtgggaa ccgccctgat caagaagtac ccaaagctgg
     4381 aaagcgagtt cgtgtacgga gactacaagg tctacgacgt gcgcaagatg attgccaaat
     4441 ctgagcagga gatcggaaag gccaccgcaa agtacttctt ctacagcaac atcatgaatt
     4501 tcttcaagac cgaaatcacc cttgcaaacg gtgagatccg gaagaggccg ctcatcgaga
     4561 ctaatgggga gactggcgaa atcgtgtggg acaagggcag agatttcgct accgtgcgca
     4621 aagtgctttc tatgcctcaa gtgaacatcg tgaagaaaac cgaggtgcaa accggaggct
     4681 tttctaagga atcaatcctc cccaagcgca actccgacaa gctcattgca aggaagaagg
     4741 attgggaccc taagaagtac ggcggattcg attcaccaac tgtggcttat tctgtcctgg
     4801 tcgtggctaa ggtggaaaaa ggaaagtcta agaagctcaa gagcgtgaag gaactgctgg
     4861 gtatcaccat tatggagcgc agctccttcg agaagaaccc aattgacttt ctcgaagcca
     4921 aaggttacaa ggaagtcaag aaggacctta tcatcaagct cccaaagtat agcctgttcg
     4981 aactggagaa tgggcggaag cggatgctcg cctccgctgg cgaacttcag aagggtaatg
     5041 agctggctct cccctccaag tacgtgaatt tcctctacct tgcaagccat tacgagaagc
     5101 tgaaggggag ccccgaggac aacgagcaaa agcaactgtt tgtggagcag cataagcatt
     5161 atctggacga gatcattgag cagatttccg agttttctaa acgcgtcatt ctcgctgatg
     5221 ccaacctcga taaagtcctt agcgcataca ataagcacag agacaaacca attcgggagc
     5281 aggctgagaa tatcatccac ctgttcaccc tcaccaatct tggtgcccct gccgcattca
     5341 agtacttcga caccaccatc gaccggaaac gctatacctc caccaaagaa gtgctggacg
     5401 ccaccctcat ccaccagagc atcaccggac tttacgaaac tcggattgac ctctcacagc
     5461 tcggagggga tgagggagct cccaagaaaa agcgcaaggt aggtagttcc taatctagat
     5521 gagggtcccc acctgggacc cttgagagta tcaggtctcc cacgtgggag acaagaaatc
     5581 cctgtttaat atttaaacag cagtgttccc catctgggtc cttgcacccc tcactctggc
     5641 ctcagccgac tgcacagcgg cccctgcatc cccttggctg tgaggcccct ggacaagcag
     5701 aggtggccag agctgggagg catggccctg gggtcccacg aatttgctgg ggaatctcgt
     5761 ttttcttctt aagacttttg ggacatggtt tgactcccga acatcaccga cgtgtctcct
     5821 gtttttctgg gtggcctcgg gacacctgcc ctgcccccac gagggtcagg actgtgactc
     5881 tttttagggc caggcaggtg cctggacatt tgccttgctg gacggggact ggggatgtgg
     5941 gagggagcag acaggaggaa tcatgtcagg cctgtgtgtg aaaggaagct ccactgtcac
     6001 cctccacctc ttcacccccc actcaccagt gtcccctcca ctgtcacatt gtaactgaac
     6061 ttcaggataa taaagtgttt gcctccaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     6121 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     6181 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     6241 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     6301 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     6361 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaccga attcactggc cgtcgtttta
     6421 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     6481 cctttcgcca gatcaagctt atcgataccg tcgacctcga gggggggccc ggtaccagct
     6541 tttgttccct ttagtgaggg ttaatttcga gcttggcgta atcatggtca tagctgtttc
     6601 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6661 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6721 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6781 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6841 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6901 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6961 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7021 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7081 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7141 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7201 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7261 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7321 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7381 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     7441 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7501 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7561 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7621 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7681 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7741 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7801 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7861 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7921 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7981 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     8041 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     8101 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     8161 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     8221 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     8281 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     8341 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8401 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8461 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8521 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8581 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8641 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8701 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     8761 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     8821 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     8881 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     8941 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     9001 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     9061 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     9121 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     9181 taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     9241 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     9301 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgta
     9361 atacgactca ctatagggcg aattggagct ccaccgcggt ggcggccgct ctagcctcga
     9421 ggctagaact agtggatctc gagccccagc tggttctttc cgcctcagaa gccatagagc
     9481 ccaccgcatc cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc
     9541 ccaccccacc ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat
     9601 tttattagga aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg
     9661 ggcaaacaac agatggctgg caactagaag gcacagtcga ggctgatcag cgagctctac
     9721 gatgcatggg gtcgtgcgct cctttcggtc gggcgctgcg ggtcgtgggg cgggcgtcag
     9781 gcaccgggct tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg
     9841 gtgacggtga agccgagccg ctcgtagaag gggaggttgc ggggcgcgga ggtctccagg
     9901 aaggcgggca ccccggcgcg ctcggccgcc tccactccgg ggagcacgac ggcgctgccc
     9961 agacccttgc cctggtggtc gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc
    10021 ttgggccggt gcggcgccag gaggccttcc atctgttgct gcgcggccag ccgggaaccg
    10081 ctcaactcgg ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc gacgctctcc
    10141 ggcgtggtcc agaccgccac cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg
    10201 agcccgacgc gcgtgaggaa gagttcttgc agctcggtga cccgctcgat gtggcggtcc
    10261 ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg
    10321 gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc ggtcatggta
    10381 agctggtcga aaggcccgga gatgaggaag aggagaacag cgcggcagac gtgcgctttt
    10441 gaagcgtgca gaatgccggg cctccggagg accttcgggc gcccgccccg cccctgagcc
    10501 cgcccctgag cccgcccccg gacccacccc ttcccagcct ctgagcccag aaagcgaagg
    10561 agcaaagctg ctattggccg ctgccccaaa ggcctacccg cttccattgc tcagcggtgc
    10621 tgtccatctg cacgagacta gtgagacgtg ctacttccat ttgtcacgtc ctgcacgacg
    10681 cgagctgcgg ggcggggggg aacttcctga ctaggggagg agtagaaggt ggcgcgaagg
    10741 ggccaccaaa gaacggagcc ggttggcgcc taccggtgga tgtggaatgt gtgcgaggcc
    10801 agaggccact tgtgtagcgc caagtgccca gcggggctgc taaagcgcat gctccagact
    10861 gccttgggaa aagcgcctcc cctacccggt agaattcgat ctggcacgac aggtttcccg
    10921 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
    10981 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
    11041 aatttcacac aggaaacagc tatgaccatg attacgccaa gcttaattcc ctccccagca
    11101 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
    11161 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
    11221 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
    11281 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tttgcaaaga tggataaagt
    11341 tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa
    11401 tt
//
