LbCpf1-2NLS

LOCUS       LbCpf1-2NLS               10514 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  bacterial expression of LbCpf1-2NLS.
ACCESSION   .
VERSION     .
KEYWORDS    LbCpf1-2NLS
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10514)
  AUTHORS   Moreno-Mateos MA, Fernandez JP, Rouet R, Vejnar CE, Lane MA, Mis E, 
            Khokha MK, Doudna JA, Giraldez AJ
  TITLE     CRISPR-Cpf1 mediates efficient homology-directed repair and 
            temperature-controlled genome editing.
  JOURNAL   Nat Commun. 2017 Dec 8;8(1):2024. doi: 10.1038/s41467-017-01836-2.
  PUBMED    29222508
REFERENCE   2  (bases 1 to 10514)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10514
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     506..525
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     609..628
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        609..627
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             716..738
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             758..775
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             785..1885
                     /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     1859..1882
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1940..1960
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1970..5653
                     /codon_start=1
                     /product="CRISPR-associated protein from Lachnospiraceae 
                     bacterium ND2006 (Zetsche et al., 2015)"
                     /label=LbCpf1
                     /translation="MSKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAE
                     DYKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKE
                     IAKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFDNRENMFS
                     EEAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFEG
                     EFFNFVLTQEGIDVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVL
                     SDRESLSFYGEGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKN
                     GPAISTISKDIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLEQ
                     LQEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKD
                     LLDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYS
                     KDKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVNG
                     NYEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKL
                     IDFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLV
                     EEGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLKK
                     EELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKI
                     NTEVRVLLKHDDNPYVIGIDRGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTD
                     YHSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNSG
                     FKNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQ
                     NGFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALD
                     YKNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQ
                     QGDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSRN
                     YEAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTS
                     VKH"
     CDS             5657..5683
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5702..5722
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5729..5749
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5789..6493
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(5931..5949)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6199..6218
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             6500..6520
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6652..6670)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6666..6713
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(7079..7107)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(7134..7152)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7220..7324
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7325..8185
                     /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(7543..7562)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8356..8944
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8845..8864
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9098..9115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    9130..9269
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(9195..9214)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9355..9377
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(9371..9562)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
       61 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg
      121 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc
      181 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
      241 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
      301 gcatcggtcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
      361 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
      421 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
      481 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
      541 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
      601 cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta gtgccggctc
      661 cggagagctc tttaattaag cggccgccct gcaggactcg agttctagaa ataattttgt
      721 ttaactttaa gaaggagata tacatatgaa atcttctcac catcaccatc accatggttc
      781 ttctatgaaa atcgaagaag gtaaactggt aatctggatt aacggcgata aaggctataa
      841 cggtctcgct gaagtcggta agaaattcga gaaagatacc ggaattaaag tcaccgttga
      901 gcatccggat aaactggaag agaaattccc acaggttgcg gcaactggcg atggccctga
      961 cattatcttc tgggcacacg accgctttgg tggctacgct caatctggcc tgttggctga
     1021 aatcaccccg gacaaagcgt tccaggacaa gctgtatccg tttacctggg atgccgtacg
     1081 ttacaacggc aagctgattg cttacccgat cgctgttgaa gcgttatcgc tgatttataa
     1141 caaagatctg ctgccgaacc cgccaaaaac ctgggaagag atcccggcgc tggataaaga
     1201 actgaaagcg aaaggtaaga gcgcgctgat gttcaacctg caagaaccgt acttcacctg
     1261 gccgctgatt gctgctgacg ggggttatgc gttcaagtat gaaaacggca agtacgacat
     1321 taaagacgtg ggcgtggata acgctggcgc gaaagcgggt ctgaccttcc tggttgacct
     1381 gattaaaaac aaacacatga atgcagacac cgattactcc atcgcagaag ctgcctttaa
     1441 taaaggcgaa acagcgatga ccatcaacgg cccgtgggca tggtccaaca tcgacaccag
     1501 caaagtgaat tatggtgtaa cggtactgcc gaccttcaag ggtcaaccat ccaaaccgtt
     1561 cgttggcgtg ctgagcgcag gtattaacgc cgccagtccg aacaaagagc tggcaaaaga
     1621 gttcctcgaa aactatctgc tgactgatga aggtctggaa gcggttaata aagacaaacc
     1681 gctgggtgcc gtagcgctga agtcttacga ggaagagttg gcgaaagatc cacgtattgc
     1741 cgccactatg gaaaacgccc agaaaggtga aatcatgccg aacatcccgc agatgtccgc
     1801 tttctggtat gccgtgcgta ctgcggtgat caacgccgcc agcggtcgtc agactgtcga
     1861 tgaagccctg aaagacgcgc agactaattc gagctcgaac aacaacaaca ataacaataa
     1921 caacaacctc gggatcgagg aaaacctgta cttccaatcc aatgccacca tgagcaaact
     1981 ggagaagttc acgaactgtt attctctgtc gaaaacctta cgttttaagg ccattccggt
     2041 aggtaaaacg caagaaaata tcgacaataa acgtctcctg gttgaagatg aaaagcgtgc
     2101 cgaagattat aaaggtgtta aaaagctgct tgatcgctat tacctgtctt ttatcaacga
     2161 tgttctgcat tccatcaaat tgaaaaactt gaataattac atctcgttat ttcgcaaaaa
     2221 aacccgtacc gaaaaagaaa acaaggagct ggaaaacctt gaaattaatc tgcgtaaaga
     2281 aatcgcgaaa gcatttaaag gtaatgaagg atataaatct ctgtttaaaa aagacattat
     2341 tgaaaccatc ctgcctgagt tcttggatga taaagatgaa attgccctgg ttaatagctt
     2401 taatggcttt acgactgcat ttaccggctt ttttgataac cgtgaaaaca tgttcagcga
     2461 agaagcgaaa tcgacttcca ttgccttccg ttgcattaat gaaaacctga cccgctacat
     2521 ttctaatatg gacatctttg aaaaagttga tgccattttc gacaaacatg aagttcaaga
     2581 aatcaaagaa aagattttaa actccgatta tgatgttgaa gatttctttg agggagaatt
     2641 tttcaatttt gttctcaccc aagagggtat tgatgtctat aacgccatta tcggaggatt
     2701 tgttacggaa tcaggcgaaa agattaaagg actgaatgaa tacattaatt tatacaatca
     2761 gaaaaccaaa cagaaattgc caaaattcaa accactgtat aaacaggtat taagcgaccg
     2821 tgaatccttg tccttttatg gcgaaggcta tacaagcgat gaagaggtgc tcgaagtatt
     2881 ccgtaatact ttgaacaaaa actcggaaat cttctccagc attaaaaaac tggaaaaatt
     2941 gtttaaaaat ttcgatgaat atagtagcgc aggcatcttc gtcaaaaatg gccctgcgat
     3001 ttcaaccatt tctaaagaca ttttcggtga atggaacgtg atccgcgata aatggaacgc
     3061 ggagtatgac gacattcatc tgaaaaaaaa agccgttgtt accgaaaagt atgaagatga
     3121 ccgccgcaag agctttaaaa aaattggtag tttctcactg gaacagttac aagaatatgc
     3181 agacgccgat ctgtcggttg tggaaaagtt aaaagagatt attatccaga aagtcgatga
     3241 aatttataaa gtgtacggct ccagcgaaaa gctctttgat gccgactttg ttttagaaaa
     3301 atcgctgaaa aaaaacgatg ccgtggtggc catcatgaaa gatctgttgg attcggtcaa
     3361 atcgttcgaa aactatatca aggcgttctt tggcgaaggg aaagagacca atcgtgacga
     3421 atcgttctat ggcgattttg tgttggcgta tgatattctt ctgaaagtag atcacatcta
     3481 tgatgctatc cgcaactacg tgacccagaa accgtattcg aaggataaat ttaagctgta
     3541 ttttcaaaac cctcagttta tgggtggctg ggataaagat aaagaaaccg actaccgcgc
     3601 gacgatcttg cgctatggct ctaagtacta tttagccatc atggataaaa agtatgcaaa
     3661 atgcctccag aaaatcgata aggatgatgt caacgggaac tacgaaaaaa tcaactacaa
     3721 actcttgcct ggcccgaata aaatgctgcc gaaagtcttt ttttccaaaa aatggatggc
     3781 ctattataac ccttccgaag atattcagaa gatttacaag aacggtacct ttaaaaaagg
     3841 tgacatgttt aatctgaatg actgtcacaa gcttattgat ttcttcaaag atagtatttc
     3901 tcgctaccct aaatggtcaa acgcgtacga ttttaatttc tcagaaacag agaaatacaa
     3961 agatattgcg gggttttatc gcgaagtcga agaacaaggg tataaggtgt cgttcgaatc
     4021 ggcttccaaa aaggaagtgg ataaacttgt tgaagaaggc aaactctata tgttccagat
     4081 ctacaacaaa gatttctccg acaagagcca tggcacgccg aacttacata ccatgtactt
     4141 taaattactg ttcgatgaaa ataatcatgg tcagattcgc ttgagcggtg gcgccgaact
     4201 gtttatgcgc cgcgcatcat tgaaaaagga agaacttgtt gtgcacccag caaacagtcc
     4261 gattgcaaac aaaaatccgg ataacccgaa gaagactacc accttaagct atgatgtata
     4321 taaagataaa cgttttagcg aagatcagta tgaactccac attccgatcg cgattaacaa
     4381 atgtcctaag aacatcttta aaatcaatac cgaggttcgt gtactgctca aacacgacga
     4441 caacccctat gttattggca ttgaccgcgg cgaacgcaac ctgctgtata ttgtcgttgt
     4501 ggatggcaaa ggtaacattg ttgaacagta tagccttaat gaaatcatca ataatttcaa
     4561 cggcattcgt attaaaacgg actatcatag cctgctggac aaaaaagaga aagaacgctt
     4621 cgaagcccgt cagaattgga cctctatcga aaatatcaag gaactgaaag ctgggtacat
     4681 ttctcaagtg gtccacaaga tttgcgaact ggtggaaaag tatgacgcgg tgatcgcact
     4741 ggaggatctg aattcgggct ttaaaaatag ccgtgtgaag gtggagaaac aggtttacca
     4801 aaagttcgaa aaaatgctga ttgacaaact taattacatg gtggacaaaa aatctaatcc
     4861 atgtgcgacc ggcggggctc tcaaaggcta tcagattacg aataaattcg aaagcttcaa
     4921 atccatgtct acccagaacg gatttatctt ttacatcccc gcctggctga cttccaaaat
     4981 tgatccgagt accggttttg ttaatctgct gaaaaccaaa tatacgtcga tcgccgattc
     5041 aaaaaaattc attagcagct tcgaccgtat tatgtacgtc ccggaagaag acctgttcga
     5101 attcgccctt gactataaaa atttctcgcg tacggatgcc gattatatca aaaaatggaa
     5161 actgtacagc tacggcaacc gcatccgcat ttttcgcaac ccaaagaaga acaacgtatt
     5221 tgactgggag gaagtttgct tgacgagcgc ttacaaagag ctctttaaca aatatgggat
     5281 caattatcag caaggggaca tccgtgcgtt gctgtgtgag cagtcagata aagcgttcta
     5341 cagctctttc atggctttaa tgtcgctgat gttgcagatg cgcaattcga tcactggccg
     5401 caccgatgtg gatttcctga ttagcccggt caaaaatagc gatggcattt tttacgattc
     5461 acgtaactat gaagcccagg aaaacgctat tctgccgaag aacgctgatg cgaacggcgc
     5521 gtacaatatt gcccgtaagg tactgtgggc aattggtcag ttcaagaaag cggaagatga
     5581 aaagttggat aaggtcaaaa ttgcgatctc gaataaagaa tggctggaat atgcgcagac
     5641 cagcgtgaaa catgcctatc cctatgacgt gcccgattat gccagcctgg gcagcggctc
     5701 ccccaagaaa aaacgcaagg tggaagatcc taagaaaaag cggaaagtgg actaaattgg
     5761 tagtgggagc aacggcagca gcggatccgt gagcaagggc gaggaggata acatggccat
     5821 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     5881 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     5941 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     6001 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     6061 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     6121 gacccaggac tcctccctcc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
     6181 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc
     6241 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
     6301 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
     6361 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga
     6421 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
     6481 cgagctgtac aaggggtcac ctaagaaaaa gcgaaaagtt gagtaataac attggaagtg
     6541 gataacggat ccgcgatcgc ggcgcgccac ctggtggccg gccggtacca cgcgtgcgcg
     6601 ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     6661 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     6721 gaggaactat atccggatat ccacaggacg ggtgtggtcg ccatgatcgc gtagtcgata
     6781 gtggctccaa gtagcgaagc gagcaggact gggcggcggc caaagcggtc ggacagtgct
     6841 ccgagaacgg gtgcgcatag aaattgcatc aacgcatata gcgctagcag cacgccatag
     6901 tgactggcga tgctgtcgga atggacgata tcccgcaaga ggcccggcag taccggcata
     6961 accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg
     7021 ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat
     7081 taaagcttat cgatgataag ctgtcaaaca tgagaattct tgaagacgaa agggcctcgt
     7141 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     7201 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     7261 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataacatt gaaaaaggaa
     7321 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     7381 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     7441 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     7501 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     7561 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     7621 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     7681 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     7741 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     7801 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     7861 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     7921 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     7981 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     8041 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     8101 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     8161 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     8221 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     8281 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     8341 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     8401 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     8461 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     8521 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     8581 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     8641 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     8701 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     8761 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     8821 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     8881 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     8941 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     9001 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     9061 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     9121 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa
     9181 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     9241 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     9301 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     9361 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     9421 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     9481 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     9541 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     9601 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     9661 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     9721 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
     9781 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
     9841 gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg
     9901 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
     9961 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
    10021 gcacccgtgg ccaggaccca acgctgcccg agatgcgccg cgtgcggctg ctggagatgg
    10081 cggacgcgat ggatatgttc tgccaagggt tggtttgcgc attcacagtt ctccgcaaga
    10141 attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc cggcttccat
    10201 tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc agacaaggta
    10261 tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg cggcataaat
    10321 cgccgtgacg atcagcggtc caatgatcga agttaggctg gtaagagccg cgagcgatcc
    10381 ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct gcaacgcggg
    10441 catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc agcctcgcgt
    10501 cgcgaacgcc agca
//