pMBP-LbCas12a

LOCUS        pMBP-LbCas12a               9653 bp ds-DNA     circular SYN 07-JUN-2021
DEFINITION  Expresses 10xHis-MBP fused to LbCas12a.
ACCESSION   .
VERSION     .
KEYWORDS    pMBP-LbCas12a
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9653)
  AUTHORS   Chen JS, Ma E, Harrington LB, Da Costa M, Tian X, Palefsky JM, 
            Doudna JA
  TITLE     CRISPR-Cas12a target binding unleashes indiscriminate 
            single-stranded DNase activity.
  JOURNAL   Science. 2018 Apr 27;360(6387):436-439. doi: 
            10.1126/science.aar6245. Epub 2018 Feb 15.
  PUBMED    29449511
REFERENCE   2  (bases 1 to 9653)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 7, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9653
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     990..1009
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1093..1112
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1093..1111
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1200..1222
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1242..1271
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1281..2381
                     /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     2355..2378
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             2436..2456
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2463..6146
                     /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"
     primer_bind     complement(6275..6293)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6289..6336
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(6702..6730)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(6757..6775)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6843..6947
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6948..7808
                     /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(7166..7185)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7979..8567
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8468..8487
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8721..8738
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8753..8892
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8818..8837)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8978..9000
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8994..9185)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ccaggaccca acgctgcccg agatgcgccg cgtgcggctg ctggagatgg cggacgcgat
       61 ggatatgttc tgccaagggt tggtttgcgc attcacagtt ctccgcaaga attgattggc
      121 tccaattctt ggagtggtga atccgttagc gaggtgccgc cggcttccat tcaggtcgag
      181 gtggcccggc tccatgcacc gcgacgcaac gcggggaggc agacaaggta tagggcggcg
      241 cctacaatcc atgccaaccc gttccatgtg ctcgccgagg cggcataaat cgccgtgacg
      301 atcagcggtc caatgatcga agttaggctg gtaagagccg cgagcgatcc ttgaagctgt
      361 ccctgatggt cgtcatctac ctgcctggac agcatggcct gcaacgcggg catcccgatg
      421 ccgccggaag cgagaagaat cataatgggg aaggccatcc agcctcgcgt cgcgaacgcc
      481 agcaagacgt agcccagcgc gtcggccgcc atgccggcga taatggcctg cttctcgccg
      541 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
      601 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
      661 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
      721 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
      781 aagggcatcg gtcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
      841 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
      901 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
      961 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
     1021 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
     1081 atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc tctagtgccg
     1141 gctccggaga gctctttaat taagcggccg ccctgcagga ctcgagttct agaaataatt
     1201 ttgtttaact ttaagaagga gatatacata tgaaatcttc tcaccatcac catcaccatc
     1261 accatcacca tggttcttct atgaaaatcg aagaaggtaa actggtaatc tggattaacg
     1321 gcgataaagg ctataacggt ctcgctgaag tcggtaagaa attcgagaaa gataccggaa
     1381 ttaaagtcac cgttgagcat ccggataaac tggaagagaa attcccacag gttgcggcaa
     1441 ctggcgatgg ccctgacatt atcttctggg cacacgaccg ctttggtggc tacgctcaat
     1501 ctggcctgtt ggctgaaatc accccggaca aagcgttcca ggacaagctg tatccgttta
     1561 cctgggatgc cgtacgttac aacggcaagc tgattgctta cccgatcgct gttgaagcgt
     1621 tatcgctgat ttataacaaa gatctgctgc cgaacccgcc aaaaacctgg gaagagatcc
     1681 cggcgctgga taaagaactg aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag
     1741 aaccgtactt cacctggccg ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa
     1801 acggcaagta cgacattaaa gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga
     1861 ccttcctggt tgacctgatt aaaaacaaac acatgaatgc agacaccgat tactccatcg
     1921 cagaagctgc ctttaataaa ggcgaaacag cgatgaccat caacggcccg tgggcatggt
     1981 ccaacatcga caccagcaaa gtgaattatg gtgtaacggt actgccgacc ttcaagggtc
     2041 aaccatccaa accgttcgtt ggcgtgctga gcgcaggtat taacgccgcc agtccgaaca
     2101 aagagctggc aaaagagttc ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg
     2161 ttaataaaga caaaccgctg ggtgccgtag cgctgaagtc ttacgaggaa gagttggcga
     2221 aagatccacg tattgccgcc actatggaaa acgcccagaa aggtgaaatc atgccgaaca
     2281 tcccgcagat gtccgctttc tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg
     2341 gtcgtcagac tgtcgatgaa gccctgaaag acgcgcagac taattcgagc tcgaacaaca
     2401 acaacaataa caataacaac aacctcggga tcgaggaaaa cctgtacttc caatccaatg
     2461 caatgagcaa actggagaag ttcacgaact gttattctct gtcgaaaacc ttacgtttta
     2521 aggccattcc ggtaggtaaa acgcaagaaa atatcgacaa taaacgtctc ctggttgaag
     2581 atgaaaagcg tgccgaagat tataaaggtg ttaaaaagct gcttgatcgc tattacctgt
     2641 cttttatcaa cgatgttctg cattccatca aattgaaaaa cttgaataat tacatctcgt
     2701 tatttcgcaa aaaaacccgt accgaaaaag aaaacaagga gctggaaaac cttgaaatta
     2761 atctgcgtaa agaaatcgcg aaagcattta aaggtaatga aggatataaa tctctgttta
     2821 aaaaagacat tattgaaacc atcctgcctg agttcttgga tgataaagat gaaattgccc
     2881 tggttaatag ctttaatggc tttacgactg catttaccgg cttttttgat aaccgtgaaa
     2941 acatgttcag cgaagaagcg aaatcgactt ccattgcctt ccgttgcatt aatgaaaacc
     3001 tgacccgcta catttctaat atggacatct ttgaaaaagt tgatgccatt ttcgacaaac
     3061 atgaagttca agaaatcaaa gaaaagattt taaactccga ttatgatgtt gaagatttct
     3121 ttgagggaga atttttcaat tttgttctca cccaagaggg tattgatgtc tataacgcca
     3181 ttatcggagg atttgttacg gaatcaggcg aaaagattaa aggactgaat gaatacatta
     3241 atttatacaa tcagaaaacc aaacagaaat tgccaaaatt caaaccactg tataaacagg
     3301 tattaagcga ccgtgaatcc ttgtcctttt atggcgaagg ctatacaagc gatgaagagg
     3361 tgctcgaagt attccgtaat actttgaaca aaaactcgga aatcttctcc agcattaaaa
     3421 aactggaaaa attgtttaaa aatttcgatg aatatagtag cgcaggcatc ttcgtcaaaa
     3481 atggccctgc gatttcaacc atttctaaag acattttcgg tgaatggaac gtgatccgcg
     3541 ataaatggaa cgcggagtat gacgacattc atctgaaaaa aaaagccgtt gttaccgaaa
     3601 agtatgaaga tgaccgccgc aagagcttta aaaaaattgg tagtttctca ctggaacagt
     3661 tacaagaata tgcagacgcc gatctgtcgg ttgtggaaaa gttaaaagag attattatcc
     3721 agaaagtcga tgaaatttat aaagtgtacg gctccagcga aaagctcttt gatgccgact
     3781 ttgttttaga aaaatcgctg aaaaaaaacg atgccgtggt ggccatcatg aaagatctgt
     3841 tggattcggt caaatcgttc gaaaactata tcaaggcgtt ctttggcgaa gggaaagaga
     3901 ccaatcgtga cgaatcgttc tatggcgatt ttgtgttggc gtatgatatt cttctgaaag
     3961 tagatcacat ctatgatgct atccgcaact acgtgaccca gaaaccgtat tcgaaggata
     4021 aatttaagct gtattttcaa aaccctcagt ttatgggtgg ctgggataaa gataaagaaa
     4081 ccgactaccg cgcgacgatc ttgcgctatg gctctaagta ctatttagcc atcatggata
     4141 aaaagtatgc aaaatgcctc cagaaaatcg ataaggatga tgtcaacggg aactacgaaa
     4201 aaatcaacta caaactcttg cctggcccga ataaaatgct gccgaaagtc tttttttcca
     4261 aaaaatggat ggcctattat aacccttccg aagatattca gaagatttac aagaacggta
     4321 cctttaaaaa aggtgacatg tttaatctga atgactgtca caagcttatt gatttcttca
     4381 aagatagtat ttctcgctac cctaaatggt caaacgcgta cgattttaat ttctcagaaa
     4441 cagagaaata caaagatatt gcggggtttt atcgcgaagt cgaagaacaa gggtataagg
     4501 tgtcgttcga atcggcttcc aaaaaggaag tggataaact tgttgaagaa ggcaaactct
     4561 atatgttcca gatctacaac aaagatttct ccgacaagag ccatggcacg ccgaacttac
     4621 ataccatgta ctttaaatta ctgttcgatg aaaataatca tggtcagatt cgcttgagcg
     4681 gtggcgccga actgtttatg cgccgcgcat cattgaaaaa ggaagaactt gttgtgcacc
     4741 cagcaaacag tccgattgca aacaaaaatc cggataaccc gaagaagact accaccttaa
     4801 gctatgatgt atataaagat aaacgtttta gcgaagatca gtatgaactc cacattccga
     4861 tcgcgattaa caaatgtcct aagaacatct ttaaaatcaa taccgaggtt cgtgtactgc
     4921 tcaaacacga cgacaacccc tatgttattg gcattgaccg cggcgaacgc aacctgctgt
     4981 atattgtcgt tgtggatggc aaaggtaaca ttgttgaaca gtatagcctt aatgaaatca
     5041 tcaataattt caacggcatt cgtattaaaa cggactatca tagcctgctg gacaaaaaag
     5101 agaaagaacg cttcgaagcc cgtcagaatt ggacctctat cgaaaatatc aaggaactga
     5161 aagctgggta catttctcaa gtggtccaca agatttgcga actggtggaa aagtatgacg
     5221 cggtgatcgc actggaggat ctgaattcgg gctttaaaaa tagccgtgtg aaggtggaga
     5281 aacaggttta ccaaaagttc gaaaaaatgc tgattgacaa acttaattac atggtggaca
     5341 aaaaatctaa tccatgtgcg accggcgggg ctctcaaagg ctatcagatt acgaataaat
     5401 tcgaaagctt caaatccatg tctacccaga acggatttat cttttacatc cccgcctggc
     5461 tgacttccaa aattgatccg agtaccggtt ttgttaatct gctgaaaacc aaatatacgt
     5521 cgatcgccga ttcaaaaaaa ttcattagca gcttcgaccg tattatgtac gtcccggaag
     5581 aagacctgtt cgaattcgcc cttgactata aaaatttctc gcgtacggat gccgattata
     5641 tcaaaaaatg gaaactgtac agctacggca accgcatccg catttttcgc aacccaaaga
     5701 agaacaacgt atttgactgg gaggaagttt gcttgacgag cgcttacaaa gagctcttta
     5761 acaaatatgg gatcaattat cagcaagggg acatccgtgc gttgctgtgt gagcagtcag
     5821 ataaagcgtt ctacagctct ttcatggctt taatgtcgct gatgttgcag atgcgcaatt
     5881 cgatcactgg ccgcaccgat gtggatttcc tgattagccc ggtcaaaaat agcgatggca
     5941 ttttttacga ttcacgtaac tatgaagccc aggaaaacgc tattctgccg aagaacgctg
     6001 atgcgaacgg cgcgtacaat attgcccgta aggtactgtg ggcaattggt cagttcaaga
     6061 aagcggaaga tgaaaagttg gataaggtca aaattgcgat ctcgaataaa gaatggctgg
     6121 aatatgcgca gaccagcgtg aaacattaat aacattggaa gtggataacg gatccgcgat
     6181 cgcggcgcgc cacctggtgg ccggccggta ccacgcgtgc gcgctgatcc ggctgctaac
     6241 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     6301 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
     6361 tatccacagg acgggtgtgg tcgccatgat cgcgtagtcg atagtggctc caagtagcga
     6421 agcgagcagg actgggcggc ggccaaagcg gtcggacagt gctccgagaa cgggtgcgca
     6481 tagaaattgc atcaacgcat atagcgctag cagcacgcca tagtgactgg cgatgctgtc
     6541 ggaatggacg atatcccgca agaggcccgg cagtaccggc ataaccaagc ctatgcctac
     6601 agcatccagg gtgacggtgc cgaggatgac gatgagcgca ttgttagatt tcatacacgg
     6661 tgcctgactg cgttagcaat ttaactgtga taaactaccg cattaaagct tatcgatgat
     6721 aagctgtcaa acatgagaat tcttgaagac gaaagggcct cgtgatacgc ctatttttat
     6781 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     6841 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6901 gacaataacc ctgataaatg cttcaataac attgaaaaag gaagagtatg agtattcaac
     6961 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     7021 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     7081 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     7141 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     7201 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     7261 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     7321 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     7381 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     7441 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     7501 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7561 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7621 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     7681 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7741 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7801 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7861 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7921 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7981 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     8041 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     8101 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     8161 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     8221 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     8281 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     8341 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     8401 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     8461 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8521 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8581 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     8641 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     8701 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     8761 ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac
     8821 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     8881 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     8941 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     9001 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     9061 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     9121 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     9181 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     9241 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
     9301 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
     9361 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
     9421 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
     9481 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
     9541 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
     9601 cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg tgg
//