LOCUS       sequence-214627-        9758 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial vector for expression of Snap-tagged Staphylococcus aureus
            dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9758)
  AUTHORS   Savic N, Ringnalda FC, Lindsay H, Berk C, Bargsten K, Li Y, Neri D, 
            Robinson MD, Ciaudo C, Hall J, Jinek M, Schwank G
  TITLE     Covalent linkage of the DNA repair template to the CRISPR-Cas9 
            nuclease enhances homology-directed repair.
  JOURNAL   Elife. 2018 May 29;7. pii: 33761. doi: 10.7554/eLife.33761.
  PUBMED    29809142
REFERENCE   2  (bases 1 to 9758)
  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"; journalName: "Elife."; 
            date: "2018-05-29"; volume: "7. pii: 33761. doi"; pages: " 
            10.7554/eLife.33761"
COMMENT     Sequence Label: pNS38-SadCas9-SNAP
FEATURES             Location/Qualifiers
     source          1..9758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(18..36)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        104..208
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(209..277,278..1069)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(427..446)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1240..1828
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1729..1748
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1982..1999
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2014..2156
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2082..2101)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2242..2264
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2258..2449)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3907..3926
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4010..4029
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4010..4028
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             4117..4139
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4159..4176
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4186..5286
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     5260..5283
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             5299..5319
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5323..8481
                     /codon_start=1
                     /product="Cas9 endonuclease from the Staphylococcus aureus 
                     Type II CRISPR/Cas system"
                     /label=SaCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MKRNYILGLAIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNE
                     GRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEE
                     FSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGE
                     VRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFG
                     WKDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKF
                     QIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKE
                     IIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKA
                     INLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSI
                     KVINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENA
                     KYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVK
                     QEEASKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRF
                     SVQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKER
                     NKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEI
                     FITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKD
                     NDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKK
                     DNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKN
                     LDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLL
                     NRIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQ
                     IIKKG"
     CDS             8515..8541
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             8521..8547
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             8560..8580
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8587..8607
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8641..9183
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="optimized for bacterial expression"
                     /translation="DKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVEV
                     PAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLKV
                     VKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLAV
                     KEWLLAHEGHRLGKPGLG"
     CDS             9199..9219
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(9294..9312)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      9308..9355
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(9721..9726,9727..9743,9744..9749))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
       61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      181 gcttcaataa cattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
      481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
      541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
      601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
      661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
      721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
      781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
      841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
      901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
      961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     1981 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2041 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc
     2101 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     2161 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     2221 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     2281 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     2341 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     2401 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     2461 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
     2521 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
     2581 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
     2641 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
     2701 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
     2761 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
     2821 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
     2881 tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga
     2941 tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg
     3001 tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc
     3061 cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg
     3121 acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt
     3181 ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccag tgatcgaagt
     3241 taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg
     3301 cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat
     3361 aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc
     3421 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt
     3481 gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat
     3541 cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg
     3601 tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg
     3661 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gacgctctcc
     3721 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
     3781 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     3841 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     3901 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     3961 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc
     4021 actataggga gaccacaacg gtttccctct agtgccggct ccggagagct ctttaattaa
     4081 gcggccgccc tgcaggactc gagttctaga aataattttg tttaacttta agaaggagat
     4141 atacatatga aatcttctca ccatcaccat caccatggtt cttctatgaa aatcgaagaa
     4201 ggtaaactgg taatctggat taacggcgat aaaggctata acggtctcgc tgaagtcggt
     4261 aagaaattcg agaaagatac cggaattaaa gtcaccgttg agcatccgga taaactggaa
     4321 gagaaattcc cacaggttgc ggcaactggc gatggccctg acattatctt ctgggcacac
     4381 gaccgctttg gtggctacgc tcaatctggc ctgttggctg aaatcacccc ggacaaagcg
     4441 ttccaggaca agctgtatcc gtttacctgg gatgccgtac gttacaacgg caagctgatt
     4501 gcttacccga tcgctgttga agcgttatcg ctgatttata acaaagatct gctgccgaac
     4561 ccgccaaaaa cctgggaaga gatcccggcg ctggataaag aactgaaagc gaaaggtaag
     4621 agcgcgctga tgttcaacct gcaagaaccg tacttcacct ggccgctgat tgctgctgac
     4681 gggggttatg cgttcaagta tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat
     4741 aacgctggcg cgaaagcggg tctgaccttc ctggttgacc tgattaaaaa caaacacatg
     4801 aatgcagaca ccgattactc catcgcagaa gctgccttta ataaaggcga aacagcgatg
     4861 accatcaacg gcccgtgggc atggtccaac atcgacacca gcaaagtgaa ttatggtgta
     4921 acggtactgc cgaccttcaa gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca
     4981 ggtattaacg ccgccagtcc gaacaaagag ctggcaaaag agttcctcga aaactatctg
     5041 ctgactgatg aaggtctgga agcggttaat aaagacaaac cgctgggtgc cgtagcgctg
     5101 aagtcttacg aggaagagtt ggcgaaagat ccacgtattg ccgccactat ggaaaacgcc
     5161 cagaaaggtg aaatcatgcc gaacatcccg cagatgtccg ctttctggta tgccgtgcgt
     5221 actgcggtga tcaacgccgc cagcggtcgt cagactgtcg atgaagccct gaaagacgcg
     5281 cagactaatg ggatcgagga aaacctgtac ttccaatcca atatgaaaag gaattatatc
     5341 ttaggattag ctatcggaat tacatcagtg ggttatggaa ttattgatta tgaaactaga
     5401 gatgtcatag atgcgggcgt acgtttattt aaagaggcta atgttgaaaa taatgaagga
     5461 cgacgatcaa aaagaggtgc cagaaggctt aagaggcgtc gtagacatag aatacaaaga
     5521 gtaaagaaac ttttatttga ttacaatttg ttgacagatc atagtgagct aagtggaatc
     5581 aatccttacg aggcgcgcgt aaagggatta agtcaaaaat taagtgaaga ggaattttct
     5641 gcggcattgc tacatttagc aaagcgtaga ggtgtacata atgttaatga agtggaagaa
     5701 gatacaggta atgaattatc cactaaagaa caaatttcaa gaaatagtaa agcgttagaa
     5761 gagaagtatg ttgcagaatt acagttggaa cgtttgaaaa aagacggtga agtgagaggt
     5821 tcgattaacc gtttcaaaac atctgactat gtaaaagaag caaagcagtt attaaaagta
     5881 caaaaagcat atcatcaact tgatcaatca tttatagaca cttatattga tttattggaa
     5941 acaagaagaa catattatga gggaccaggt gaaggtagcc catttggatg gaaagatatt
     6001 aaagaatggt atgaaatgtt aatgggacat tgtacgtatt tcccagaaga attacgtagt
     6061 gtgaaatatg cctataatgc tgatttatat aatgcgctga atgatttgaa caacttggtt
     6121 attacacgag atgagaatga gaagctagag tattatgaaa aattccaaat tatcgagaat
     6181 gtctttaaac aaaagaaaaa gccgacgctt aaacaaattg cgaaggaaat cttggtgaat
     6241 gaagaagaca tcaaaggcta tcgtgtcaca agtacaggta aaccagaatt tacaaacttg
     6301 aaagtttatc acgatatcaa agatattaca gcaagaaaag aaattatcga gaatgcagag
     6361 ctactcgatc aaatagctaa aatattaact atttaccagt catcagaaga tatacaagaa
     6421 gaattaacaa acctaaattc agaattgaca caagaagaga ttgaacaaat ttcaaacttg
     6481 aaaggttata caggaactca taacctttca ctaaaggcaa taaatttaat attagacgaa
     6541 ttgtggcata cgaacgataa tcaaatagct attttcaatc gtttgaaact tgtacctaaa
     6601 aaggtagatt taagccaaca aaaagaaatt cctactactt tagttgatga ttttatactg
     6661 tctccagtag tgaaacgttc atttatacaa tctattaaag ttattaacgc tattattaaa
     6721 aaatacggtt tgccaaatga tattattatt gaacttgcga gagaaaagaa ttctaaagat
     6781 gcacaaaaaa tgattaatga aatgcagaag agaaatcgtc aaacgaatga acgtattgag
     6841 gaaattataa gaacgacagg taaagaaaat gctaaatatt taattgaaaa aattaagctg
     6901 cacgatatgc aagaagggaa atgtttatac tcgttagaag caatccctct agaagattta
     6961 cttaataatc cattcaatta cgaagtagac catatcattc cacgttctgt ttctttcgat
     7021 aactctttca ataataaagt gttggtgaaa caagaagaag ctagtaaaaa aggtaataga
     7081 acgccatttc aatatttaag ttcttcagat tctaaaataa gttatgagac attcaaaaag
     7141 catattttaa atcttgctaa aggcaaaggt agaatctcta agacgaaaaa agaatatttg
     7201 ttagaagaac gagatatcaa tcgcttcagt gtccaaaaag attttattaa ccgtaactta
     7261 gtagatacac gctatgcgac aagagggtta atgaacttat taagatctta ttttagagtg
     7321 aataacttag atgtcaaagt gaaatcgatt aatggcggat tcacaagttt cttaagaagg
     7381 aaatggaagt tcaaaaaaga aagaaataag ggctacaaac accatgctga agatgcactg
     7441 attattgcga acgctgattt tattttcaaa gaatggaaaa aactagataa agctaaaaaa
     7501 gtgatggaaa atcaaatgtt tgaagaaaag caagctgaaa gtatgcctga aattgagact
     7561 gagcaagagt ataaagaaat ttttataacg cctcatcaaa ttaaacatat taaggatttt
     7621 aaagattata aatattcaca tagagttgat aaaaagccga atagagagtt aataaatgat
     7681 acattatatt ctacgagaaa agatgacaag ggtaatacat taatcgttaa taacttaaat
     7741 ggtttatacg ataaagataa tgataaattg aaaaaattaa ttaataaatc acctgaaaaa
     7801 ttattgatgt atcatcatga tccacaaaca tatcaaaaat taaaattgat catggaacaa
     7861 tatggcgatg agaaaaatcc gctttataaa tattatgaag aaacaggcaa ttacttaaca
     7921 aaatatagta aaaaagataa cggaccagtc atcaaaaaaa ttaaatatta tggtaacaag
     7981 ctaaatgcgc atttagatat tacggatgat tatccaaata gcagaaataa agtagtaaaa
     8041 ctttcattaa aaccatatcg ctttgatgtt tatttagata atggggtata taaatttgtg
     8101 acagttaaaa atttagatgt tatcaaaaaa gaaaactact atgaagttaa ttcaaagtgt
     8161 tatgaagaag caaaaaaact gaagaaaatt agtaatcaag cagaatttat cgcaagtttt
     8221 tacaataatg acttgattaa gattaacgga gaattatata gagtcatagg tgtaaataat
     8281 gatctactta acagaattga agtaaatatg atagacatca catatagaga atatttagag
     8341 aacatgaatg ataaaagacc acctagaata attaaaacaa tagcaagcaa aacacaatct
     8401 attaaaaagt attctacaga tattctaggc aatctttatg aagtgaagag taaaaagcat
     8461 cctcaaatca taaagaaagg atacgaacat tggaagtgga taacggatcc agcctatccc
     8521 tatgacgtgc ccgattatgc cagcctgggc agcggctccc ccaagaaaaa acgcaaggtg
     8581 gaagatccta agaaaaagcg gaaagtggac ggcattggta gtgggagcaa cggcagcagc
     8641 gacaaagatt gcgaaatgaa acgtaccacc ctggatagcc cgctgggcaa actggaactg
     8701 agcggctgcg aacagggcct gcatgaaatt aaactgctgg gtaaaggcac cagcgcggcc
     8761 gatgcggttg aagttccggc cccggccgcc gtgctgggtg gtccggaacc gctgatgcag
     8821 gcgaccgcgt ggctgaacgc gtattttcat cagccggaag cgattgaaga atttccggtt
     8881 ccggcgctgc atcatccggt gtttcagcag gagagcttta cccgtcaggt gctgtggaaa
     8941 ctgctgaaag tggttaaatt tggcgaagtg attagctatc agcagctggc ggccctggcg
     9001 ggtaatccgg cggccaccgc cgccgttaaa accgcgctga gcggtaaccc ggtgccgatt
     9061 ctgattccgt gccatcgtgt ggttagctct agcggtgcgg ttggcggtta tgaaggtggt
     9121 ctggcggtga aagagtggct gctggcccat gaaggtcatc gtctgggtaa accgggtctg
     9181 ggacctgcag gcgggtcacc taagaaaaaa cgaaaagttg agtaaggtac cacgcgtgcg
     9241 cgctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     9301 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     9361 aggaggaact atatccggat atccacagga cgggtgtggt cgccatgatc gcgtagtcga
     9421 tagtggctcc aagtagcgaa gcgagcagga ctgggcggcg gccaaagcgg tcggacagtg
     9481 ctccgagaac gggtgcgcat agaaattgca tcaacgcata tagcgctagc agcacgccat
     9541 agtgactggc gatgctgtcg gaatggacga tatcccgcaa gaggcccggc agtaccggca
     9601 taaccaagcc tatgcctaca gcatccaggg tgacggtgcc gaggatgacg atgagcgcat
     9661 tgttagattt catacacggt gcctgactgc gttagcaatt taactgtgat aaactaccgc
     9721 attaaagctt atcgatgata agctgtcaaa catgagaa
//
