p32Lb

LOCUS       p32Lb      15453 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Construct conventional Cpf1 vectors that uses U3p and UBI10p to 
            express single crRNA (or PTC/CA) and FnCpf1, respectively. For plant
            genome editing. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15453)
  AUTHORS   Ding D, Chen K, Chen Y, Li H, Xie K
  TITLE     Engineering Introns to Express RNA Guides for Cas9- and 
            Cpf1-Mediated Multiplex Genome Editing.
  JOURNAL   Mol Plant. 2018 Apr 2;11(4):542-552. doi: 
            10.1016/j.molp.2018.02.005. Epub 2018 Feb 17.
  PUBMED    29462720
REFERENCE   2  (bases 1 to 15453)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.molp.2018.02.005"; journalName: "Mol Plant"; date: 
            "2018-04-2- 2"; volume: "11"; issue: "4"; pages: "542-552"
COMMENT     Sequence Label: p32Lb
FEATURES             Location/Qualifiers
     source          1..15453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    116..137
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(152..157,158..175,176..182)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    190..206
                     /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     195..217
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     214..230
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     214..230
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        248..628
                     /label=OsU3 promoter
                     /note="Oryza sativa (rice) snRNA U3 promoter"
     misc_RNA        645..720
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        801..2510
                     /gene="RUBQ2"
                     /label=Ubi promoter
                     /note="promoter for the rice (Oryza sativa) polyubiquitin 
                     gene RUBQ2"
     intron          1549..2510
                     /note="intron upstream of the coding sequence for the rice 
                     polyubiquitin gene RUBQ2"
     protein_bind    2521..2545
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2602..6285
                     /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             6286..6333
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6340..6420
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     protein_bind    complement(6453..6477)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6488..6511
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      6533..6785
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(6794..6811)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6794..6810)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6803..6825)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    7013..7037
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             8338..8967
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             9396..10469
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      10535..10729
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(10965..10987)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    11073..11213
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     11128..11147
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(11228..11245)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11399..11987)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11479..11498)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(12074..12868)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    13293..13317
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    13395..13569
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(13610..14635)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     promoter        complement(14702..15378)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     complement(14745..14765)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
ORIGIN
        1 gcaagctgct ctagccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
       61 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
      121 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
      181 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac atgattacgc
      241 caagcttaag gaatctttaa acatacgaac agatcactta aagttcttct gaagcaactt
      301 aaagttatca ggcatgcatg gatcttggag gaatcagatg tgcagtcagg gaccatagca
      361 caagacaggc gtcttctact ggtgctacca gcaaatgctg gaagccggga acactgggta
      421 cgttggaaac cacgtgatgt gaagaagtaa gataaactgt aggagaaaag catttcgtag
      481 tgggccatga agcctttcag gacatgtatt gcagtatggg ccggcccatt acgcaattgg
      541 acgacaacaa agactagtat tagtaccacc tcggctatcc acatagatca aagctgattt
      601 aaaagagttg tgcagatgat ccgtggcagg agaccgaggt ctcggtttta gagctagaaa
      661 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
      721 ttttttgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt tttagcgcgt
      781 gcatgcctgc aggtccacaa attcgggtca aggcggaagc cagcgcgcca ccccacgtca
      841 gcaaatacgg aggcgcgggg ttgacggcgt cacccggtcc taacggcgac caacaaacca
      901 gccagaagaa attacagtaa aaaaaaagta aattgcactt tgatccacct tttattacct
      961 aagtctcaat ttggatcacc cttaaaccta tcttttcaat ttgggccggg ttgtggtttg
     1021 gactaccatg aacaactttt cgtcatgtct aacttccctt tcagcaaaca tatgaaccat
     1081 atatagagga gatcggccgt atactagagc tgatgtgttt aaggtcgttg attgcacgag
     1141 aaaaaaaaat ccaaatcgca acaatagcaa atttatctgg ttcaaagtga aaagatatgt
     1201 ttaaaggtag tccaaagtaa aacttataga taataaaatg tggtccaaag cgtaattcac
     1261 tcaaaaaaaa tcaacgagac gtgtaccaaa cggagacaaa cggcatcttc tcgaaatttc
     1321 ccaaccgctc gctcgcccgc ctcgtcttcc cggaaaccgc ggtggtttca gcgtggcgga
     1381 ttctccaagc agacggagac gtcacggcac gggactcctc ccaccaccca accgccataa
     1441 ataccagccc cctcatctcc tctcctcgca tcagctccac ccccgaaaaa tttctcccca
     1501 atctcgcgag gctctcgtcg tcgaatcgaa tcctctcgcg tcctcaaggt acgctgcttc
     1561 tcctctcctc gcttcgtttc gattcgattt cggacgggtg aggttgtttt gttgctagat
     1621 ccgattggtg gttagggttg tcgatgtgat tatcgtgaga tgtttagggg ttgtagatct
     1681 gatggttgtg atttgggcac ggttggttcg ataggtggaa tcgtggttag gttttgggat
     1741 tggatgttgg ttctgatgat tggggggaat ttttacggtt agatgaattg ttggatgatt
     1801 cgattgggga aatcggtgta gatctgttgg ggaattgtgg aactagtcat gcctgagtga
     1861 ttggtgcgat ttgtagcgtg ttccatcttg taggccttgt tgcgagcatg ttcagatcta
     1921 ctgttccgct cttgattgag ttattggtgc catgggttgg tgcaaacaca ggctttaata
     1981 tgttatatct gttttgtgtt tgatgtagat ctgtagggta gttcttctta gacatggttc
     2041 aattatgtag cttgtgcgtt tcgatttgat ttcatatgtt cacagattag ataatgatga
     2101 actcttttaa ttaattgtca atggtaaata ggaagtcttg tcgctatatc tgtcataatg
     2161 atctcatgtt actatctgcc agtaatttat gctaagaact atattagaat atcatgttac
     2221 aatctgtagt aatatcatgt tacaatctgt agttcatcta tataatctat tgtggtaatt
     2281 tctttttact atctgtgtga agattattgc cactagttca ttctacttat ttctgaagtt
     2341 caggatacgt gtgctgttac tacctatctg aatacatgtg tgatgtgcct gttactatct
     2401 ttttgaatac atgtatgttc tgttggaata tgtttgctgt ttgatccgtt gttgtgtcct
     2461 taatcttgtg ctagttctta ccctatctgt ttggtgatta tttcttgcag atagttatca
     2521 acaagtttgt acaaaaaagc aggcttcgaa ggagatagaa ccaattctct aaggaaatac
     2581 ttaaccatgg gtaccgccac catgagcaag ctggagaagt ttacaaactg ctactccctg
     2641 tctaagaccc tgaggttcaa ggccatccct gtgggcaaga cccaggagaa catcgacaat
     2701 aagcggctgc tggtggagga cgagaagaga gccgaggatt ataagggcgt gaagaagctg
     2761 ctggatcgct actatctgtc ttttatcaac gacgtgctgc acagcatcaa gctgaagaat
     2821 ctgaacaatt acatcagcct gttccggaag aaaaccagaa ccgagaagga gaataaggag
     2881 ctggagaacc tggagatcaa tctgcggaag gagatcgcca aggccttcaa gggcaacgag
     2941 ggctacaagt ccctgtttaa gaaggatatc atcgagacaa tcctgccaga gttcctggac
     3001 gataaggacg agatcgccct ggtgaacagc ttcaatggct ttaccacagc cttcaccggc
     3061 ttctttgata acagagagaa tatgttttcc gaggaggcca agagcacatc catcgccttc
     3121 aggtgtatca acgagaatct gacccgctac atctctaata tggacatctt cgagaaggtg
     3181 gacgccatct ttgataagca cgaggtgcag gagatcaagg agaagatcct gaacagcgac
     3241 tatgatgtgg aggatttctt tgagggcgag ttctttaact ttgtgctgac acaggagggc
     3301 atcgacgtgt ataacgccat catcggcggc ttcgtgaccg agagcggcga gaagatcaag
     3361 ggcctgaacg agtacatcaa cctgtataat cagaaaacca agcagaagct gcctaagttt
     3421 aagccactgt ataagcaggt gctgagcgat cgggagtctc tgagcttcta cggcgagggc
     3481 tatacatccg atgaggaggt gctggaggtg tttagaaaca ccctgaacaa gaacagcgag
     3541 atcttcagct ccatcaagaa gctggagaag ctgttcaaga attttgacga gtactctagc
     3601 gccggcatct ttgtgaagaa cggccccgcc atcagcacaa tctccaagga tatcttcggc
     3661 gagtggaacg tgatccggga caagtggaat gccgagtatg acgatatcca cctgaagaag
     3721 aaggccgtgg tgaccgagaa gtacgaggac gatcggagaa agtccttcaa gaagatcggc
     3781 tccttttctc tggagcagct gcaggagtac gccgacgccg atctgtctgt ggtggagaag
     3841 ctgaaggaga tcatcatcca gaaggtggat gagatctaca aggtgtatgg ctcctctgag
     3901 aagctgttcg acgccgattt tgtgctggag aagagcctga agaagaacga cgccgtggtg
     3961 gccatcatga aggacctgct ggattctgtg aagagcttcg agaattacat caaggccttc
     4021 tttggcgagg gcaaggagac aaacagggac gagtccttct atggcgattt tgtgctggcc
     4081 tacgacatcc tgctgaaggt ggaccacatc tacgatgcca tccgcaatta tgtgacccag
     4141 aagccctact ctaaggataa gttcaagctg tattttcaga accctcagtt catgggcggc
     4201 tgggacaagg ataaggagac agactatcgg gccaccatcc tgagatacgg ctccaagtac
     4261 tatctggcca tcatggataa gaagtacgcc aagtgcctgc agaagatcga caaggacgat
     4321 gtgaacggca attacgagaa gatcaactat aagctgctgc ccggccctaa taagatgctg
     4381 ccaaaggtgt tcttttctaa gaagtggatg gcctactata accccagcga ggacatccag
     4441 aagatctaca agaatggcac attcaagaag ggcgatatgt ttaacctgaa tgactgtcac
     4501 aagctgatcg acttctttaa ggatagcatc tcccggtatc caaagtggtc caatgcctac
     4561 gatttcaact tttctgagac agagaagtat aaggacatcg ccggctttta cagagaggtg
     4621 gaggagcagg gctataaggt gagcttcgag tctgccagca agaaggaggt ggataagctg
     4681 gtggaggagg gcaagctgta tatgttccag atctataaca aggacttttc cgataagtct
     4741 cacggcacac ccaatctgca caccatgtac ttcaagctgc tgtttgacga gaacaatcac
     4801 ggacagatca ggctgagcgg aggagcagag ctgttcatga ggcgcgcctc cctgaagaag
     4861 gaggagctgg tggtgcaccc agccaactcc cctatcgcca acaagaatcc agataatccc
     4921 aagaaaacca caaccctgtc ctacgacgtg tataaggata agaggttttc tgaggaccag
     4981 tacgagctgc acatcccaat cgccatcaat aagtgcccca agaacatctt caagatcaat
     5041 acagaggtgc gcgtgctgct gaagcacgac gataacccct atgtgatcgg catcgatagg
     5101 ggcgagcgca atctgctgta tatcgtggtg gtggacggca agggcaacat cgtggagcag
     5161 tattccctga acgagatcat caacaacttc aacggcatca ggatcaagac agattaccac
     5221 tctctgctgg acaagaagga gaaggagagg ttcgaggccc gccagaactg gacctccatc
     5281 gagaatatca aggagctgaa ggccggctat atctctcagg tggtgcacaa gatctgcgag
     5341 ctggtggaga agtacgatgc cgtgatcgcc ctggaggacc tgaactctgg ctttaagaat
     5401 agccgcgtga aggtggagaa gcaggtgtat cagaagttcg agaagatgct gatcgataag
     5461 ctgaactaca tggtggacaa gaagtctaat ccttgtgcaa caggcggcgc cctgaagggc
     5521 tatcagatca ccaataagtt cgagagcttt aagtccatgt ctacccagaa cggcttcatc
     5581 ttttacatcc ctgcctggct gacatccaag atcgatccat ctaccggctt tgtgaacctg
     5641 ctgaaaacca agtataccag catcgccgat tccaagaagt tcatcagctc ctttgacagg
     5701 atcatgtacg tgcccgagga ggatctgttc gagtttgccc tggactataa gaacttctct
     5761 cgcacagacg ccgattacat caagaagtgg aagctgtact cctacggcaa ccggatcaga
     5821 atcttccgga atcctaagaa gaacaacgtg ttcgactggg aggaggtgtg cctgaccagc
     5881 gcctataagg agctgttcaa caagtacggc atcaattatc agcagggcga tatcagagcc
     5941 ctgctgtgcg agcagtccga caaggccttc tactctagct ttatggccct gatgagcctg
     6001 atgctgcaga tgcggaacag catcacaggc cgcaccgacg tggattttct gatcagccct
     6061 gtgaagaact ccgacggcat cttctacgat agccggaact atgaggccca ggagaatgcc
     6121 atcctgccaa agaacgccga cgccaatggc gcctataaca tcgccagaaa ggtgctgtgg
     6181 gccatcggcc agttcaagaa ggccgaggac gagaagctgg ataaggtgaa gatcgccatc
     6241 tctaacaagg agtggctgga gtacgcccag accagcgtga agcacaaaag gccggcggcc
     6301 acgaaaaagg ccggccaggc aaaaaagaaa aagggatcct acccatacga tgttccagat
     6361 tacgcttatc cctacgacgt gcctgattat gcatacccat atgatgtccc cgactatgcc
     6421 taagaattcg cggccgcact cgagatatct agacccagct ttcttgtaca aagtggttga
     6481 taacagcgac tacaaggatg acgatgacaa ggcttagagc tcgaatttcc ccgatcgttc
     6541 aaacatttgg caataaagtt tcttaagatt gaatcctgtt gccggtcttg cgatgattat
     6601 catataattt ctgttgaatt acgttaagca tgtaataatt aacatgtaat gcatgacgtt
     6661 atttatgaga tgggttttta tgattagagt cccgcaatta tacatttaat acgcgataga
     6721 aaacaaaata tagcgcgcaa actaggataa attatcgcgc gcggtgtcat ctatgttact
     6781 agatcgggaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     6841 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     6901 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgc tagagcagct
     6961 tgagcttgga tcagattgtc gtttcccgcc ttcagtttaa actatcagtg tttgacagga
     7021 tatattggcg ggtaaaccta agagaaaaga gcgtttatta gaataatcgg atatttaaaa
     7081 gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc atgccaacca cagggttccc
     7141 ctcgggatca aagtactttg atccaacccc tccgctgcta tagtgcagtc ggcttctgac
     7201 gttcagtgca gccgtcttct gaaaacgaca tgtcgcacaa gtcctaagtt acgcgacagg
     7261 ctgccgccct gcccttttcc tggcgttttc ttgtcgcgtg ttttagtcgc ataaagtaga
     7321 atacttgcga ctagaaccgg agacattacg ccatgaacaa gagcgccgcc gctggcctgc
     7381 tgggctatgc ccgcgtcagc accgacgacc aggacttgac caaccaacgg gccgaactgc
     7441 acgcggccgg ctgcaccaag ctgttttccg agaagatcac cggcaccagg cgcgaccgcc
     7501 cggagctggc caggatgctt gaccacctac gccctggcga cgttgtgaca gtgaccaggc
     7561 tagaccgcct ggcccgcagc acccgcgacc tactggacat tgccgagcgc atccaggagg
     7621 ccggcgcggg cctgcgtagc ctggcagagc cgtgggccga caccaccacg ccggccggcc
     7681 gcatggtgtt gaccgtgttc gccggcattg ccgagttcga gcgttcccta atcatcgacc
     7741 gcacccggag cgggcgcgag gccgccaagg cccgaggcgt gaagtttggc ccccgcccta
     7801 ccctcacccc ggcacagatc gcgcacgccc gcgagctgat cgaccaggaa ggccgcaccg
     7861 tgaaagaggc ggctgcactg cttggcgtgc atcgctcgac cctgtaccgc gcacttgagc
     7921 gcagcgagga agtgacgccc accgaggcca ggcggcgcgg tgccttccgt gaggacgcat
     7981 tgaccgaggc cgacgccctg gcggccgccg agaatgaacg ccaagaggaa caagcatgaa
     8041 accgcaccag gacggccagg acgaaccgtt tttcattacc gaagagatcg aggcggagat
     8101 gatcgcggcc gggtacgtgt tcgagccgcc cgcgcacgtc tcaaccgtgc ggctgcatga
     8161 aatcctggcc ggtttgtctg atgccaagct ggcggcctgg ccggccagct tggccgctga
     8221 agaaaccgag cgccgccgtc taaaaaggtg atgtgtattt gagtaaaaca gcttgcgtca
     8281 tgcggtcgct gcgtatatga tgcgatgagt aaataaacaa atacgcaagg ggaacgcatg
     8341 aaggttatcg ctgtacttaa ccagaaaggc gggtcaggca agacgaccat cgcaacccat
     8401 ctagcccgcg ccctgcaact cgccggggcc gatgttctgt tagtcgattc cgatccccag
     8461 ggcagtgccc gcgattgggc ggccgtgcgg gaagatcaac cgctaaccgt tgtcggcatc
     8521 gaccgcccga cgattgaccg cgacgtgaag gccatcggcc ggcgcgactt cgtagtgatc
     8581 gacggagcgc cccaggcggc ggacttggct gtgtccgcga tcaaggcagc cgacttcgtg
     8641 ctgattccgg tgcagccaag cccttacgac atatgggcca ccgccgacct ggtggagctg
     8701 gttaagcagc gcattgaggt cacggatgga aggctacaag cggcctttgt cgtgtcgcgg
     8761 gcgatcaaag gcacgcgcat cggcggtgag gttgccgagg cgctggccgg gtacgagctg
     8821 cccattcttg agtcccgtat cacgcagcgc gtgagctacc caggcactgc cgccgccggc
     8881 acaaccgttc ttgaatcaga acccgagggc gacgctgccc gcgaggtcca ggcgctggcc
     8941 gctgaaatta aatcaaaact catttgagtt aatgaggtaa agagaaaatg agcaaaagca
     9001 caaacacgct aagtgccggc cgtccgagcg cacgcagcag caaggctgca acgttggcca
     9061 gcctggcaga cacgccagcc atgaagcggg tcaactttca gttgccggcg gaggatcaca
     9121 ccaagctgaa gatgtacgcg gtacgccaag gcaagaccat taccgagctg ctatctgaat
     9181 acatcgcgca gctaccagag taaatgagca aatgaataaa tgagtagatg aattttagcg
     9241 gctaaaggag gcggcatgga aaatcaagaa caaccaggca ccgacgccgt ggaatgcccc
     9301 atgtgttgag gaacgggcgg ttggccaggc gtaagcggct gggttgcctg ccggccctgc
     9361 aatggcactg gaacccccaa gcccgaggaa tcggcgtgag cggtcgcaaa ccatccggcc
     9421 cggtacaaat cggcgcggcg ctgggtgatg acctggtgga gaagttgaag gccgcgcagg
     9481 ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg tgaatcgtgg caagcggccg
     9541 ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc cggtgcgccg tcgattagga
     9601 agccgcccaa gggcgacgag caaccagatt ttttcgttcc gatgctctat gacgtgggca
     9661 cccgcgatag tcgcagcatc atggacgtgg ccgttttccg tctgtcgaag cgtgaccgac
     9721 gagctggcga ggtgatccgc tacgagcttc cagacgggca cgtagaggtt tccgcagggc
     9781 cggccggcat ggccagtgtg tgggattacg acctggtact gatggcggtt tcccatctaa
     9841 ccgaatccat gaaccgatac cgggaaggga agggagacaa gcccggccgc gtgttccgtc
     9901 cacacgttgc ggacgtactc aagttctgcc ggcgagccga tggcggaaag cagaaagacg
     9961 acctggtaga aacctgcatt cggttaaaca ccacgcacgt tgccatgcag cgtacgaaga
    10021 aggccaagaa cggccgcctg gtgacggtat ccgagggtga agccttgatt agccgctaca
    10081 agatcgtaaa gagcgaaacc gggcggccgg agtacatcga gatcgagcta gctgattgga
    10141 tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct gacggttcac cccgattact
    10201 ttttgatcga tcccggcatc ggccgttttc tctaccgcct ggcacgccgc gccgcaggca
    10261 aggcagaagc cagatggttg ttcaagacga tctacgaacg cagtggcagc gccggagagt
    10321 tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc aaatgacctg ccggagtacg
    10381 atttgaagga ggaggcgggg caggctggcc cgatcctagt catgcgctac cgcaacctga
    10441 tcgagggcga agcatccgcc ggttcctaat gtacggagca gatgctaggg caaattgccc
    10501 tagcagggga aaaaggtcga aaagcactct ttcctgtgaa tagcacgtac attgggaacc
    10561 caaagccgta cattgggaac cggaacccgt acattgggaa cccaaagccg tacattggga
    10621 accggtcaca catgtaagtg actgatataa aagagaaaaa aggcgatttt tccgcctaaa
    10681 actctttaaa acttattaaa actcttaaaa cccgcctggc ctgtgcataa ctgtctggcc
    10741 agcgcacagc cgaagagctg caaaaagcgc ctacccttcg gtcgctgcgc tccctacgcc
    10801 ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc aaaaatggct ggcctacggc
    10861 caggcaatct accagggcgc ggacaagccg cgccgtcgcc actcgaccgc cggcgcccac
    10921 atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
    10981 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
    11041 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
    11101 agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
    11161 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
    11221 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
    11281 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
    11341 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
    11401 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
    11461 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
    11521 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
    11581 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
    11641 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
    11701 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
    11761 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
    11821 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
    11881 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
    11941 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
    12001 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
    12061 tgcattctag gtactaaaac aattcatcca gtaaaatata atattttatt ttctcccaat
    12121 caggcttgat ccccagtaag tcaaaaaata gctcgacata ctgttcttcc ccgatatcct
    12181 ccctgatcga ccggacgcag aaggcaatgt cataccactt gtccgccctg ccgcttctcc
    12241 caagatcaat aaagccactt actttgccat ctttcacaaa gatgttgctg tctcccaggt
    12301 cgccgtggga aaagacaagt tcctcttcgg gcttttccgt ctttaaaaaa tcatacagct
    12361 cgcgcggatc tttaaatgga gtgtcttctt cccagttttc gcaatccaca tcggccagat
    12421 cgttattcag taagtaatcc aattcggcta agcggctgtc taagctattc gtatagggac
    12481 aatccgatat gtcgatggag tgaaagagcc tgatgcactc cgcatacagc tcgataatct
    12541 tttcagggct ttgttcatct tcatactctt ccgagcaaag gacgccatcg gcctcactca
    12601 tgagcagatt gctccagcca tcatgccgtt caaagtgcag gacctttgga acaggcagct
    12661 ttccttccag ccatagcatc atgtcctttt cccgttccac atcataggtg gtccctttat
    12721 accggctgtc cgtcattttt aaatataggt tttcattttc tcccaccagc ttatatacct
    12781 tagcaggaga cattccttcc gtatctttta cgcagcggta tttttcgatc agttttttca
    12841 attccggtga tattctcatt ttagccattt attatttcct tcctcttttc tacagtattt
    12901 aaagataccc caagaagcta attataacaa gacgaactcc aattcactgt tccttgcatt
    12961 ctaaaacctt aaataccaga aaacagcttt ttcaaagttg ttttcaaagt tggcgtataa
    13021 catagtatcg acggagccga ttttgaaacc gcggtgatca caggcagcaa cgctctgtca
    13081 tcgttacaat caacatgcta ccctccgcga gatcatccgt gtttcaaacc cggcagctta
    13141 gttgccgttc ttccgaatag catcggtaac atgagcaaag tctgccgcct tacaacggct
    13201 ctcccgctga cgccgtcccg gactgatggg ctgcctgtat cgagtggtga ttttgtgccg
    13261 agctgccggt cggggagctg ttggctggct ggtggcagga tatattgtgg tgtaaacaaa
    13321 ttgacgctta gacaacttaa taacacattg cggacgtttt taatgtactg aattaacgcc
    13381 gaattaattc gggggatctg gattttagta ctggattttg gttttaggaa ttagaaattt
    13441 tattgataga agtattttac aaatacaaat acatactaag ggtttcttat atgctcaaca
    13501 catgagcgaa accctatagg aaccctaatt cccttatctg ggaactactc acacattatt
    13561 atggagaaac tcgagcttgt cgatcgacag atcccggtcg gcatctactc tatttctttg
    13621 ccctcggacg agtgctgggg cgtcggtttc cactatcggc gagtacttct acacagccat
    13681 cggtccagac ggccgcgctt ctgcgggcga tttgtgtacg cccgacagtc ccggctccgg
    13741 atcggacgat tgcgtcgcat cgaccctgcg cccaagctgc atcatcgaaa ttgccgtcaa
    13801 ccaagctctg atagagttgg tcaagaccaa tgcggagcat atacgcccgg agtcgtggcg
    13861 atcctgcaag ctccggatgc ctccgctcga agtagcgcgt ctgctgctcc atacaagcca
    13921 accacggcct ccagaagaag atgttggcga cctcgtattg ggaatccccg aacatcgcct
    13981 cgctccagtc aatgaccgct gttatgcggc cattgtccgt caggacattg ttggagccga
    14041 aatccgcgtg cacgaggtgc cggacttcgg ggcagtcctc ggcccaaagc atcagctcat
    14101 cgagagcctg cgcgacggac gcactgacgg tgtcgtccat cacagtttgc cagtgataca
    14161 catggggatc agcaatcgcg catatgaaat cacgccatgt agtgtattga ccgattcctt
    14221 gcggtccgaa tgggccgaac ccgctcgtct ggctaagatc ggccgcagcg atcgcatcca
    14281 tagcctccgc gaccggttgt agaacagcgg gcagttcggt ttcaggcagg tcttgcaacg
    14341 tgacaccctg tgaacggcgg gagatgcaat aggtcaggct ctcgctaaac tccccaatgt
    14401 caagcacttc cggaatcggg agcgcggccg atgcaaagtg ccgataaaca taacgatctt
    14461 tgtagaaacc atcggcgcag ctatttaccc gcaggacata tccacgccct cctacatcga
    14521 agctgaaagc acgagattct tcgccctccg agagctgcat caggtcggag acgctgtcga
    14581 acttttcgat cagaaacttc tcgacagacg tcgcggtgag ttcaggcttt ttcatatctc
    14641 attgcccccc ggatctgcga aagctcgaga gagatagatt tgtagagaga gactggtgat
    14701 ttcagcgtgt cctctccaaa tgaaatgaac ttccttatat agaggaaggg tcttgcgaag
    14761 gatagtggga ttgtgcgtca tcccttacgt cagtggagat atcacatcaa tccacttgct
    14821 ttgaagacgt ggttggaacg tcttcttttt ccacgatgct cctcgtgggt gggggtccat
    14881 ctttgggacc actgtcggca gaggcatctt gaacgatagc ctttccttta tcgcaatgat
    14941 ggcatttgta ggtgccacct tccttttcta ctgtcctttt gatgaagtga cagatagctg
    15001 ggcaatggaa tccgaggagg tttcccgata ttaccctttg ttgaaaagtc tcaatagccc
    15061 tttggtcttc tgagactgta tctttgatat tcttggagta gacgagagtg tcgtgctcca
    15121 ccatgttcac atcaatccac ttgctttgaa gacgtggttg gaacgtcttc tttttccacg
    15181 atgctcctcg tgggtggggg tccatctttg ggaccactgt cggcagaggc atcttgaacg
    15241 atagcctttc ctttatcgca atgatggcat ttgtaggtgc caccttcctt ttctactgtc
    15301 cttttgatga agtgacagat agctgggcaa tggaatccga ggaggtttcc cgatattacc
    15361 ctttgttgaa aagtctcaat agccctttgg tcttctgaga ctgtatcttt gatattcttg
    15421 gagtagacga gagtgtcgtg ctccaccatg ttg
//