LOCUS       sequence-193895-       10517 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of LbCpf1-2NLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10517)
  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 10517)
  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: "Nat 
            Commun."; date: "2017-12-8"; volume: "8(1):2024. doi"; pages: " 
            10.1038/s41467-017-01836-2"
COMMENT     Sequence Label: LbCpf1-2NLS
FEATURES             Location/Qualifiers
     source          1..10517
                     /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             5341..5361
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5371..9054
                     /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             9058..9084
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             9064..9090
                     /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             9103..9123
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9130..9150
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9190..9894
                     /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(9332..9350)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     9600..9619
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             9901..9921
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(10053..10071)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      10067..10114
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(10480..10485,10486..10502,10503..10508))
                     /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 tattgaaaaa 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 agcggtccaa 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 cagactaatt cgagctcgaa caacaacaac aataacaata acaacaacct cgggatcgag
     5341 gaaaacctgt acttccaatc caatgccacc atgagcaaac tggagaagtt cacgaactgt
     5401 tattctctgt cgaaaacctt acgttttaag gccattccgg taggtaaaac gcaagaaaat
     5461 atcgacaata aacgtctcct ggttgaagat gaaaagcgtg ccgaagatta taaaggtgtt
     5521 aaaaagctgc ttgatcgcta ttacctgtct tttatcaacg atgttctgca ttccatcaaa
     5581 ttgaaaaact tgaataatta catctcgtta tttcgcaaaa aaacccgtac cgaaaaagaa
     5641 aacaaggagc tggaaaacct tgaaattaat ctgcgtaaag aaatcgcgaa agcatttaaa
     5701 ggtaatgaag gatataaatc tctgtttaaa aaagacatta ttgaaaccat cctgcctgag
     5761 ttcttggatg ataaagatga aattgccctg gttaatagct ttaatggctt tacgactgca
     5821 tttaccggct tttttgataa ccgtgaaaac atgttcagcg aagaagcgaa atcgacttcc
     5881 attgccttcc gttgcattaa tgaaaacctg acccgctaca tttctaatat ggacatcttt
     5941 gaaaaagttg atgccatttt cgacaaacat gaagttcaag aaatcaaaga aaagatttta
     6001 aactccgatt atgatgttga agatttcttt gagggagaat ttttcaattt tgttctcacc
     6061 caagagggta ttgatgtcta taacgccatt atcggaggat ttgttacgga atcaggcgaa
     6121 aagattaaag gactgaatga atacattaat ttatacaatc agaaaaccaa acagaaattg
     6181 ccaaaattca aaccactgta taaacaggta ttaagcgacc gtgaatcctt gtccttttat
     6241 ggcgaaggct atacaagcga tgaagaggtg ctcgaagtat tccgtaatac tttgaacaaa
     6301 aactcggaaa tcttctccag cattaaaaaa ctggaaaaat tgtttaaaaa tttcgatgaa
     6361 tatagtagcg caggcatctt cgtcaaaaat ggccctgcga tttcaaccat ttctaaagac
     6421 attttcggtg aatggaacgt gatccgcgat aaatggaacg cggagtatga cgacattcat
     6481 ctgaaaaaaa aagccgttgt taccgaaaag tatgaagatg accgccgcaa gagctttaaa
     6541 aaaattggta gtttctcact ggaacagtta caagaatatg cagacgccga tctgtcggtt
     6601 gtggaaaagt taaaagagat tattatccag aaagtcgatg aaatttataa agtgtacggc
     6661 tccagcgaaa agctctttga tgccgacttt gttttagaaa aatcgctgaa aaaaaacgat
     6721 gccgtggtgg ccatcatgaa agatctgttg gattcggtca aatcgttcga aaactatatc
     6781 aaggcgttct ttggcgaagg gaaagagacc aatcgtgacg aatcgttcta tggcgatttt
     6841 gtgttggcgt atgatattct tctgaaagta gatcacatct atgatgctat ccgcaactac
     6901 gtgacccaga aaccgtattc gaaggataaa tttaagctgt attttcaaaa ccctcagttt
     6961 atgggtggct gggataaaga taaagaaacc gactaccgcg cgacgatctt gcgctatggc
     7021 tctaagtact atttagccat catggataaa aagtatgcaa aatgcctcca gaaaatcgat
     7081 aaggatgatg tcaacgggaa ctacgaaaaa atcaactaca aactcttgcc tggcccgaat
     7141 aaaatgctgc cgaaagtctt tttttccaaa aaatggatgg cctattataa cccttccgaa
     7201 gatattcaga agatttacaa gaacggtacc tttaaaaaag gtgacatgtt taatctgaat
     7261 gactgtcaca agcttattga tttcttcaaa gatagtattt ctcgctaccc taaatggtca
     7321 aacgcgtacg attttaattt ctcagaaaca gagaaataca aagatattgc ggggttttat
     7381 cgcgaagtcg aagaacaagg gtataaggtg tcgttcgaat cggcttccaa aaaggaagtg
     7441 gataaacttg ttgaagaagg caaactctat atgttccaga tctacaacaa agatttctcc
     7501 gacaagagcc atggcacgcc gaacttacat accatgtact ttaaattact gttcgatgaa
     7561 aataatcatg gtcagattcg cttgagcggt ggcgccgaac tgtttatgcg ccgcgcatca
     7621 ttgaaaaagg aagaacttgt tgtgcaccca gcaaacagtc cgattgcaaa caaaaatccg
     7681 gataacccga agaagactac caccttaagc tatgatgtat ataaagataa acgttttagc
     7741 gaagatcagt atgaactcca cattccgatc gcgattaaca aatgtcctaa gaacatcttt
     7801 aaaatcaata ccgaggttcg tgtactgctc aaacacgacg acaaccccta tgttattggc
     7861 attgaccgcg gcgaacgcaa cctgctgtat attgtcgttg tggatggcaa aggtaacatt
     7921 gttgaacagt atagccttaa tgaaatcatc aataatttca acggcattcg tattaaaacg
     7981 gactatcata gcctgctgga caaaaaagag aaagaacgct tcgaagcccg tcagaattgg
     8041 acctctatcg aaaatatcaa ggaactgaaa gctgggtaca tttctcaagt ggtccacaag
     8101 atttgcgaac tggtggaaaa gtatgacgcg gtgatcgcac tggaggatct gaattcgggc
     8161 tttaaaaata gccgtgtgaa ggtggagaaa caggtttacc aaaagttcga aaaaatgctg
     8221 attgacaaac ttaattacat ggtggacaaa aaatctaatc catgtgcgac cggcggggct
     8281 ctcaaaggct atcagattac gaataaattc gaaagcttca aatccatgtc tacccagaac
     8341 ggatttatct tttacatccc cgcctggctg acttccaaaa ttgatccgag taccggtttt
     8401 gttaatctgc tgaaaaccaa atatacgtcg atcgccgatt caaaaaaatt cattagcagc
     8461 ttcgaccgta ttatgtacgt cccggaagaa gacctgttcg aattcgccct tgactataaa
     8521 aatttctcgc gtacggatgc cgattatatc aaaaaatgga aactgtacag ctacggcaac
     8581 cgcatccgca tttttcgcaa cccaaagaag aacaacgtat ttgactggga ggaagtttgc
     8641 ttgacgagcg cttacaaaga gctctttaac aaatatggga tcaattatca gcaaggggac
     8701 atccgtgcgt tgctgtgtga gcagtcagat aaagcgttct acagctcttt catggcttta
     8761 atgtcgctga tgttgcagat gcgcaattcg atcactggcc gcaccgatgt ggatttcctg
     8821 attagcccgg tcaaaaatag cgatggcatt ttttacgatt cacgtaacta tgaagcccag
     8881 gaaaacgcta ttctgccgaa gaacgctgat gcgaacggcg cgtacaatat tgcccgtaag
     8941 gtactgtggg caattggtca gttcaagaaa gcggaagatg aaaagttgga taaggtcaaa
     9001 attgcgatct cgaataaaga atggctggaa tatgcgcaga ccagcgtgaa acatgcctat
     9061 ccctatgacg tgcccgatta tgccagcctg ggcagcggct cccccaagaa aaaacgcaag
     9121 gtggaagatc ctaagaaaaa gcggaaagtg gactaaattg gtagtgggag caacggcagc
     9181 agcggatccg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     9241 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     9301 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     9361 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     9421 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     9481 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctc
     9541 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     9601 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     9661 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     9721 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     9781 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     9841 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caaggggtca
     9901 cctaagaaaa aacgaaaagt tgagtaataa cattggaagt ggataacgga tccgcgatcg
     9961 cggcgcgcca cctggtggcc ggccggtacc acgcgtgcgc gctgatccgg ctgctaacaa
    10021 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
    10081 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata
    10141 tccacaggac gggtgtggtc gccatgatcg cgtagtcgat agtggctcca agtagcgaag
    10201 cgagcaggac tgggcggcgg ccaaagcggt cggacagtgc tccgagaacg ggtgcgcata
    10261 gaaattgcat caacgcatat agcgctagca gcacgccata gtgactggcg atgctgtcgg
    10321 aatggacgat atcccgcaag aggcccggca gtaccggcat aaccaagcct atgcctacag
    10381 catccagggt gacggtgccg aggatgacga tgagcgcatt gttagatttc atacacggtg
    10441 cctgactgcg ttagcaattt aactgtgata aactaccgca ttaaagctta tcgatgataa
    10501 gctgtcaaac atgagaa
//
