LOCUS       sequence-307947-       19232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 2 cloning vector for a single guide RNA, already contains 
            Cas9, Bar selection cassette and FAST marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 19232)
  AUTHORS   Grutzner R, Martin P, Horn C, Mortensen S, Cram EJ, Lee-Parsons CWT,
            Stuttmann J, Marillonnet S
  TITLE     High-efficiency genome editing in plants mediated by a Cas9 gene 
            containing multiple introns.
  JOURNAL   Plant Commun. 2020 Nov 23;2(2):100135. doi: 
            10.1016/j.xplc.2020.100135. eCollection 2021 Mar 8.
  PUBMED    33898975
REFERENCE   2  (bases 1 to 19232)
  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: "Plant 
            Commun."; date: "2020-11-23"; volume: "2(2):100135. doi"; pages: " 
            10.1016/j.xplc.2020.100135. eCollection 2021 Mar 8"
COMMENT     Sequence Label: pAGM55285
FEATURES             Location/Qualifiers
     source          1..19232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    36..60
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        4788..4892
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4893..5687
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      5858..6446
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6347..6366
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6582..6606
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        6771..6950
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    6986..7041
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     terminator      7602..8309
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     CDS             10012..10032
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             15765..15785
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      15812..16064
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    16250..16271
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(16286..16291,16292..16309,16310..16316)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    16324..16340
                     /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     16329..16351
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     16348..16364
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     16348..16364
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             16360..16683
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    16377..16433
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(16434..16451)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(16434..16450)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(16443..16465)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(16659..16678)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_RNA        16780..16855
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             18067..18762
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSDMALK
                     LVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHK"
ORIGIN
        1 gggacacgaa gtgatccgtt taaactatca gtgtttgaca ggatatattg gcgggtaaac
       61 ctaagagaaa agagcgttta ttagaataat cggatattta aaagggcgtg aaaaggttta
      121 tccgttcgtc catttgtatg tgcatgccaa ccacagggtt cccctcggga gtgccgcaag
      181 ggtgacttcg cctatattgg acgacggcgc gcagagggcg acctcttttt gggttacgat
      241 tgtaggatta tcactaaaac aatacatgaa catattcaaa tggcaatctc tctaaggcat
      301 tggaaataaa tacaaataac agttgggtgg agtttttcga cctgagggcg ttaaccttct
      361 gttaacctaa aagctcttgc ccaaacagca gaatcggcgc taattgccag cggcggaact
      421 tttccagttt cgcgaaaaat atcgccactg gcaaggaatg ggtttgagat ggcgaagtct
      481 gtcctaaaag cagcgcctgt agttgtaggg ttgacggcct tgatggagcg tcatgccgat
      541 gccctctcga gccaacttca agcacatcat cttaaggttt tcccgccgca ttccgagaag
      601 ggcattcgaa cattcgggcc atcggaggcg tccaagctgc tcggcgttgg cgagtcatat
      661 ttacggcaga ccgcgtctga gatgccagag ttgaatgtta gcatgagccc gggtggcagg
      721 cgaatgttct caattgaaga tatccatgtg attcggaagt atatggatca ggtcggccgc
      781 gggaaccggc gctacctgcc acatcgtcga ggcggcgagc agcttcaggt tatctctgtg
      841 atgaatttca aaggtgggtc gggtaagacc accaccgccg cgcatctggc gcagtacctc
      901 gctatgcgcg gatatcgagt cttggccatt gatctcgatc ctcaagcgag cctttctgca
      961 ctctttggga gccaaccgga aacggacgtt ggcccgaacg aaacgctcta cggcgctata
     1021 aggtatgatg atgagcaggt ggcaatcgaa cgagtcgtcc gagggactta cattcccgac
     1081 ctccacctga ttcctggtaa ccttgagctg atggagtttg aacacgatac gccacgcgcg
     1141 ctgatgaacc gcaaagaggg cgacacgctc ttttatggtc gcatcagcca agtaattgaa
     1201 gatatcgcgg ataactatga cgtcgtggtc atcgactgcc ctccccagct tgggtatctc
     1261 acgctatccg cattgactgc ggcgacgtcc attcttgtca cggtccatcc gcagatgctg
     1321 gatgtgatgt cgatgaacca gtttctggca atgacatcga accttttgcg tgaaatcgag
     1381 aatgctggcg ccaagttcaa gtttaattgg atgcgctatc tgataacccg tttcgaaccg
     1441 agcgacggac cacagaacca aatggtaggt tatctgcggt cgatttttgg cgaaaatgtc
     1501 ctcaattttc cgatgcttaa aaccaccgcg gtttcggacg ctggcctgac aaaccagact
     1561 ctattcgaag tggagcgtgg cctgttcacg cgctcgacct atgatcgagc cttggaggcg
     1621 atgaacgccg tcaacgacga gatcgaaaca ctgatcaaaa aagcatgggg taggcccaca
     1681 tgagccggaa gcacatcctt ggcgtgtcaa ctgacgcccc tgaaacgtcg cccgccgaca
     1741 ataggacggc aaagaaccgc tccatgccgc tcctcggcgt aacaaggaag gagcgcgatc
     1801 cggcaacgaa gctcacagcg aacattggta acgcactgcg agagcaaaac gatcgtctta
     1861 gccgtgccga agagatcgag cggcgactcg ctgaaggtca ggcagtgata gagttggatg
     1921 cctcgtcaat agaaccgtct ttcgtgcagg atcgtatgcg aggggacatt gacgggctcc
     1981 ttacttcgat ccgggaacaa ggacagcaag tcccaatcct tgtgcgaccg catccgagcc
     2041 agccgggccg atatcaggtt gccttcggcc accgccggct acgcgccgtt tcagaactcg
     2101 gacttccggt cagagcggtc gttcgcgaac tgacggacga gcaagtggtc gtagcacagg
     2161 gtcaggaaaa caatgagcgc gaagatctta ccttcatcga aaaggcgcgc ttcgcacatc
     2221 gcctgaacag gcagttttct cgagagattg tcatcgccgc gatgtcgatc gacaagagca
     2281 atttgtccaa gatgcttctg ctcgtcgacg ccctcccctc tgaactgacc gatgctattg
     2341 gtgccgctcc tggtgttgga cggccgagtt ggcaacaact tgccgagctg attgagaaag
     2401 tttcttcacc ggccgacgtg gctaaatatg ctatgtcgga ggaagttcaa gcgctgccat
     2461 cggcagaacg attcaaggcg gtgatcgcta gtctgaagcc cagtcgggtt gcgcgtggac
     2521 ttcccgaggt catggccacc ccagacggca ccagaattgc acaggtgacg cagagcaagg
     2581 ccaaactgga aatcacgatt gacaggaagg cgacgcccga ttttgcgacc ttcgtgctcg
     2641 atcatgtgcc agcgctgtat caagcgtacc acgctgagaa ccaacggaaa cggggagagt
     2701 aaaccgcaaa agaaaagagc cccctcaacg tcgccgtcgc ggaagccctt ctgtctctct
     2761 agcgcgaaca gaatcgcatt tcctcgaatc ctcgtcaaga gtttttagcg ccgttttggt
     2821 gagctgattt cctttgcctg ctgaaaggtg aaagatgatg cagacaggaa gtgtaacgac
     2881 gccattcggg cggcggccaa tgacgcttgc gcttgtgcgg cgccagacgg cgctggccga
     2941 tatcaaacaa ggcaagacag cggacaagtg gaaggtcttt agagatgcgt ccgcggccat
     3001 ggaactactt ggaatccagt ccaacagtct tgccgtcctt gatgcgctat tgagctttca
     3061 cccggaaacg gagttgcgtc aggaggcaca gctgatcgtg ttcccgtcga atgctcagct
     3121 tgcccttcgg gcgcatggga tggctggcgc gactttgcgt aggcacatcg ccatgctcgt
     3181 ggagtcaggc ttgatcgtcc ggaaggatag cgccaacgga aagcgttacg ctcgtaagga
     3241 tggcgctggt cagatcgagc gcgcgtttgg cttcgatttg tctccgcttc tcgcgcggtc
     3301 cgaggagcta gcgatgatgg cacagcaggt gatggccgat cgagcagcat tcaggatggc
     3361 caaagaaagt ctgacgattt gccgacggga cgttcggaag ctaattacgg cagctatgga
     3421 agagggagcg gagggcgact ggcaagctgt cgaggaagtc tatgtggaac ttgtgggtag
     3481 aattccacgc gccccgacgc ttgctgatgt agagtcaatt ctcgaagaga tgtggatgct
     3541 ccaggaagag ataatcaacc ggttggaaat tagagacaat tcagaaaata atagcaccaa
     3601 tgctgcccag agcgagcagc acatacagaa ttcaaaaccc gaatccgtta atgaacttga
     3661 acctcgctct gaaaaggagc agggcgctaa gccgagtgaa atagaccggg caaggagcga
     3721 gccgataaaa gcgttccccc tcgggatgat cctgaaagca tgcccgacca ttggcaatta
     3781 tgggccgagc ggtgcggttg ctagctggcg tgacctcatg tcggctgcgg tggtggttcg
     3841 gtctatgctg ggggtcagcc cgtcggctta ccaagacgcg tgtgaggcaa tgggaccgga
     3901 gaatgcggca gcagcgatgg cgtgcatttt ggagcgagcg aacttcatca attcgcccgg
     3961 gggctatctc cgagatctga cacggcggag cgagctcggg aagttttcac ttggcccgat
     4021 gataatggcg ctcttgaagg ctagcgggca ggggacgttg cggtttggct agaattagcg
     4081 agtatggagc aggatggtct gtggtcagct gaccacagac ctaataggtt gaaaacatga
     4141 gcgttttttg gatgatcgac agaccatccg attcccggag taccaagcgt gctctgatgg
     4201 gagcgataac attactcaac aagcacgaag gccccatgcc gatcgttgat cgtgaaggag
     4261 agcctgctct acatgcggcg gtattttgcc ggccgaggca tgtagtcgcg gagcactgcc
     4321 tatttactgc cctaggcaca aacgttgact cttggatcga gctggcagac aaagcaataa
     4381 cccacacaga ggacgattaa tggctgacga agagatccag aatccgccgg acggtactgc
     4441 tgctgccgaa gttgagccgg ctgctcctag aggtagaaga gaaaagaaag caccagccga
     4501 aacagcccgc acgggatcgt tcaaatccgt gaagccgaaa acccgcggcc tcagcaaccg
     4561 agaaaaactg gagaagatcg gtcaaatcga agctcaggtc gctggcggcg caaccttgaa
     4621 ggacgccgtt aagatcgtgg gtatttccgt tcagacctat tatcaatgga agagagctgc
     4681 ggttcaacct gtctcacaga atccggccgt gtctgtttca gttgacgatg aactcggcga
     4741 gttcatccaa ctcgaggagg aaaatcggcg gctcagaaag ccgagtgcgc ggaaccccta
     4801 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4861 aaatgcttca ataatattga aaaaggaaga gtatggctaa aatgagaata tcaccggaat
     4921 tgaaaaaact gatcgaaaaa taccgctgcg taaaagatac ggaaggaatg tctcctgcta
     4981 aggtatataa gctggtggga gaaaatgaaa acctatattt aaaaatgacg gacagccggt
     5041 ataaagggac cacctatgat gtggaacggg aaaaggacat gatgctatgg ctggaaggaa
     5101 agctgcctgt tccaaaggtc ctgcactttg aacggcatga tggctggagc aatctgctca
     5161 tgagtgaggc cgatggcgtc ctttgctcgg aagagtatga agatgaacaa agccctgaaa
     5221 agattatcga gctgtatgcg gagtgcatca ggctctttca ctccatcgac atatcggatt
     5281 gtccctatac gaatagctta gacagccgct tagccgaatt ggattactta ctgaataacg
     5341 atctggccga tgtggattgc gaaaactggg aagaggacac tccatttaaa gatccgcgcg
     5401 agctgtatga ttttttaaag acggaaaagc ccgaagagga acttgtcttt tcccacggcg
     5461 acctgggaga cagcaacatc tttgtgaaag atggcaaagt aagtggcttt attgatcttg
     5521 ggagaagcgg cagggcggac aagtggtatg acattgcctt ctgcgtccgg tcgctcaggg
     5581 aggatatcgg ggaagaacag tatgtcgagc tattttttga cttactgggg atcaagcctg
     5641 attgggagaa aataaaatat tatattttac tggatgaatt gttttagctg tcagaccaag
     5701 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     5761 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     5821 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     5881 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5941 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6001 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6061 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6121 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6181 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6241 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     6301 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     6361 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     6421 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgc
     6481 tcggatctgt tggaccggac agtagtcatg gttgatgggc tgcctgtatc gagtggtgat
     6541 tttgtgccga gctgccggtc ggggagctgt tggctggctg gtggcaggat atattgtggt
     6601 gtaaacaaat tgacgcttag acaacttaat aacacattgc ggacgttttt aatgtactgg
     6661 ggttgaacac tctgtgccga attcggatcc ggagagcgga gaattaaggg agtcacgtta
     6721 tgacccccgc cgatgacgcg ggacaagccg ttttacgttt ggaactgaca gaaccgcaac
     6781 gttgaaggag ccactcagcc gcgggtttct ggagtttaat gagctaagca catacgtcag
     6841 aaaccattat tgcgcgttca aaagtcgcct aaggtcacta tcagctagca aatatttctt
     6901 gtcaaaaatg ctccactgac gttccataaa ttcccctcgg tatccaatta gagtctcata
     6961 ttcactctca tactgtattt ttacaacaat taccaacaac aacaaacaac aaacaacatt
     7021 acaattacta tttacaatta caatgtctcc tgaaagaagg cctgctgata ttagaagggc
     7081 tactgaggct gatatgcctg ctgtgtgcac tattgtgaat cattacattg agacttctac
     7141 tgtgaatttc aggactgagc ctcaagaacc acaagagtgg actgatgatc ttgtgaggct
     7201 tagagagagg tatccttggc ttgttgctga agtggatggt gaagtggctg gtattgctta
     7261 tgctggtcct tggaaggcta ggaatgctta cgattggact gctgagtcta ctgtttacgt
     7321 gtctcctagg catcaaagaa ctggtcttgg atctactctt tacactcatc ttttgaagtc
     7381 tttggaggct cagggtttca agtctgtggt tgctgtgatt ggacttccta atgatccttc
     7441 tgtgaggatg catgaggctc ttggttatgc tcctagggga atgcttagag ctgctggttt
     7501 caaacatgga aattggcatg atgtgggatt ctggcagctt gatttctctc ttcctgttcc
     7561 tcctagacct gttcttcctg tgactgagat ttgagcttgt cctgctttaa tgagatatgc
     7621 gagaagccta tgatcgcatg atatttgctt tcaattctgt tgtgcacgtt gtaaaaaacc
     7681 tgagcatgtg tagctcagat ccttaccgcc ggtttcggtt cattctaatg aatatatcac
     7741 ccgttactat cgtattttta tgaataatat tctccgttca atttactgat tgtaccctac
     7801 tacttatatg tacaatatta aaatgaaaac aatatattgt gctgaatagg tttatagcga
     7861 catctatgat agagcgccac aataacaaac aattgcgttt tattattaca aatccaattt
     7921 taaaaaaagc ggcagaaccg gtcaaaccta aaagactgat tacataaatc ttattcaaat
     7981 ttcaaaagtg ccccaggggc tagtatctac gacacaccga gcggcgaact aataacgctc
     8041 actgaaggga actccggttc cccgccggcg cgcatgggtg agattccttg aagttgagta
     8101 ttggccgtcc gctctaccga aagttacggg caccattcaa cccggtccag cacggcggcc
     8161 gggtaaccga cttgctgccc cgagaattat gcagcatttt tttggtgtat gtgggcccca
     8221 aatgaagtgc aggtcaaacc ttgacagtga cgacaaatcg ttgggcgggt ccagggcgaa
     8281 ttttgcgaca acatgtcgag gctcagcagg accgctgcaa gaattcaagc ttggagctca
     8341 acttttgatt cgctatttgc agtgcacctg tggcgttcat cacatctttt gtgacactgt
     8401 ttgcactggt cattgctatt acaaaggacc ttcctgatgt tgaaggagat cgaaagtaag
     8461 taactgcacg cataaccatt ttctttccgc tctttggctc aatccatttg acagtcaaag
     8521 acaatgttta accagctccg tttgatatat tgtctttatg tgtttgttca agcatgttta
     8581 gttaatcatg cctttgattg atcttgaata ggttccaaat atcaaccctg gcaacaaaac
     8641 ttggagtgag aaacattgca ttcctcggtt ctggacttct gctagtaaat tatgtttcag
     8701 ccatatcact agctttctac atgcctcagg tgaattcatc tatttccgtc ttaactattt
     8761 cggttaatta aagcacgaac accattactg catgtagaag cttgataaac tatcgccacc
     8821 aatttatttt tgttgcgata ttgttacttt cctcagtatg cagctttgaa aagaccaacc
     8881 ctcttatcct ttaacaatga acaggttttt agaggtagct tgatgattcc tgcacatgtg
     8941 atcttggctt caggcttaat tttccaggta aagcattatg agatactctt atatctctta
     9001 catacttttg agataatgca caagaacttc ataactatat gctttagttt ctgcatttga
     9061 cactgccaaa ttcattaatc tctaatatct ttgttgttga tctttggtag acatgggtac
     9121 tagaaaaagc aaactacacc aaggtaaaat acttttgtac aaacataaac tcgttatcac
     9181 ggaacatcaa tggagtgtat atctaacgga gtgtagaaac atttgattat tgcaggaagc
     9241 tatctcagga tattatcggt ttatatggaa tctcttctac gcagagtatc tgttattccc
     9301 cttcctctag ctttcaattt catggtgagg atatgcagtt ttctttgtat atcattcttc
     9361 ttcttctttg tagcttggag tcaaaatcgg ttccttcatg tacatacatc aaggatatgt
     9421 ccttctgaat ttttatatct tgcaataaaa atgcttgtac caattgaaac accagctttt
     9481 tgagttctat gatcactgac ttggttctaa ccaaaaaaaa aaaaatgttt aatttacata
     9541 tctaaaagta ggtttaggga aacctaaaca gtaaaatatt tgtatattat tcgaatttca
     9601 ctcatcataa aaacttaaat tgcaccataa aattttgttt tactattaat gatgtaattt
     9661 gtgtaactta agataaaaat aatattccgt aagttaaccg gctaaaacca cgtataaacc
     9721 agggaacctg ttaaaccggt tctttactgg ataaagaaat gaaagcccat gtagacagct
     9781 ccattagagc ccaaacccta aatttctcat ctatataaaa ggagtgacat tagggttttt
     9841 gttcgtcctc ttaaagcttc tcgttttctc tgccgtctct ctcattcgcg cgacgcaaac
     9901 gatcttcagg tgatcttctt tctccaaatc ctctctcata actctgattt cgtacttgtg
     9961 tatttgagct cacgctctgt ttctctcacc acagcaatgg cttctagccc accgaagaag
    10021 aagcggaagg tcagctggaa aatggacaag aagtacagca ttggacttga tattggtacg
    10081 aactcagttg ggtgggccgt tatcaccgat gaatacaagg taccttcgaa gaaatttaaa
    10141 gtgctgggca acacagatag gcacagcatt aagaagaact tgatcggagc tctgctcttt
    10201 gactctggag aaaccgcgga ggcgacaagg cttaaacgta ctgcgaggag aaggtacact
    10261 cgcaggaaga acagaatctg ttatctccaa gagatcttta gcaacgagat ggcgaaggta
    10321 aggattttta tgatatagta tgcttatgta ttttgtactg aaagcatatc ctgcttcatt
    10381 gggatattac tgaaagcatt taactacatg taaactcact tgatgatcaa taaacttgat
    10441 tttgcaggtt gacgactcgt tcttccatcg cctcgaggaa tccttcctgg tagaggaaga
    10501 taagaaacac gagcgtcacc ccatctttgg gaatattgtt gacgaagtag cctatcatga
    10561 aaagtatccg actatatacc accttcgcaa gaagctggtg gactcaaccg ataaggcaga
    10621 ccttcggctc atatacctgg ctctcgcgca catgataaag tttcgtggcc atttcttgat
    10681 cgaaggggac ctcaacccgg ataactccga tgtggataaa ctgttcattc agctcgtcca
    10741 aacctacaat cagctgttcg aggagaaccc catcaatgca tcaggtaaca ttccttagtt
    10801 acctttcttt tctttttcca tcataagttt atagattgta catgctttga gatttttctt
    10861 tgcaaacaat ctcaggtgtc gacgccaagg caatactgtc tgccagactt tcgaagtcca
    10921 gacggcttga gaatctgatc gctcaattgc caggcgagaa gaagaacggc ttgttcggga
    10981 atctgattgc actgtctctg ggcctcaccc ctaacttcaa aagcaacttt gacctcgccg
    11041 aggacgcgaa gctgcagctg tcaaaggata catacgatga tgatctggac aatctgctcg
    11101 cccaaatagg tgctcttgaa attggaactc ttcttttgtt gtctaaacct atcaatttct
    11161 ttgcggaaat ttatttgaag ctgtagagtt aaaattgagt cttttaaact tttgtaggtg
    11221 atcagtatgc cgacctgttc ttggctgcca agaatctgtc agacgctatc ttgctcagtg
    11281 acattctgcg ggtcaacacg gagataacca aagcgccact tagcgcctcc atgatcaaga
    11341 ggtacgacga gcatcaccag gatctgaccc ttctgaaggc tttggttcgc cagcaactcc
    11401 ccgagaagta caaggagatt ttctttgacc aatcgaagaa tggctacgca gggtacattg
    11461 atggaggtaa gttgttactt atgattgttt tcctctctgc tacatgtatt ttgttgttca
    11521 tttctgtaag atataagaat tgagttttcc tctgatgata ttattaggtg caagtcagga
    11581 ggaattctac aaattcatca agcctattct ggaaaagatg gacggtacag aggagctgct
    11641 cgttaaattg aaccgcgaag atttgcttcg gaagcagcgt accttcgaca atggcagcat
    11701 accgcaccag atccacctcg gtgagctgca tgctatcttg aggaggcaag aggacttcta
    11761 tccgttcctg aaagacaaca gagagaagat tgaaaagatc ctcacgttcc gcattcccta
    11821 ctatgtaggt tagtatcata tgaagaaata cctagtttca gttgatgaat gctattttct
    11881 gacctcagtt gttctctttt gagaattatt tcttttctaa tttgcctgat ttttctatta
    11941 attcattagg tccactcgca cgcgggaact cgcggtttgc gtggatgaca cgcaaatccg
    12001 aggagactat cacgccttgg aacttcgaag aggtcgtgga caagggtgcg agtgcacagt
    12061 ccttcatcga aaggatgacc aacttcgata agaatctccc aaatgagaaa gtcctgccca
    12121 agcatagtct cctgtacgaa tacttcacgg tctacaacga gctgacgaag gtgaaatatg
    12181 tgacggaggg gatgcgcaaa ccggccttcc tgtcaggtaa atcctggtcc acacttttac
    12241 gataaaaaca caagatttta aactatgaac tgatcaataa tcattcctaa aagaccacac
    12301 ttttgttttg tttctaaagt aatttttact gttataacag gtgagcagaa gaaggccatt
    12361 gtcgatctct tgttcaaaac caatcggaag gtcactgtga aacagcttaa agaggactac
    12421 tttaagaaga tcgaatgctt tgattctgtg gaaatcagcg gcgttgagga taggttcaat
    12481 gcctctcttg gcacatacca tgacctgttg aaaatcatca aggacaagga cttccttgac
    12541 aacgaggaga acgaggacat cctcgaggac atcgtgctga ctctcacgct gtttgaggac
    12601 agagaaatga tcgaggagcg ccttaagact tatgcgcatc tgttcgatga caaggtcatg
    12661 aagcagttga agaggaggag atatacaggt aagaggtcaa aaggtttccg caatgatccc
    12721 tctttttttg tttctctagt ttcaagaatt tgggtatatg actaacttct gagtgttcct
    12781 tgatgcatat ttgtgatgag acaaatgttt gttctatgtt ttaggttggg gaaggctctc
    12841 caggaagctc atcaacggca tccgcgacaa gcaatccggc aagactatac tggactttct
    12901 caaatccgac ggttttgcga atcggaactt catgcagctt attcacgatg actcactgac
    12961 cttcaaagaa gatatccaga aggcccaagt gtcaggtcag ggcgatagcc ttcacgaaca
    13021 catagccaac ctggctggat cgccagctat aaagaagggc atactgcaga cagtgaaggt
    13081 tgtggatgag ctggtgaagg taagttctgc atttggttat gctccttgca ttttaggtgt
    13141 tcgtcgcact tccatttcca tgaatagcta agattttttt tctctgcatt cattcttctt
    13201 gcctcagttc taactgtttg tggtattttt gttttaatta ttgctacagg tcatgggccg
    13261 ccataagccg gagaacatcg tcatcgagat ggcgagggaa aaccagacga ctcagaaagg
    13321 gcagaagaac tcacgggagc gcatgaagcg gatagaggaa ggcatcaagg agcttgggag
    13381 tcagattctg aaagagcacc cagtcgaaaa tactcaactc cagaacgaga agctgtacct
    13441 ctattacctc cagaatggga gagatatgta cgtcgaccaa gagctcgaca ttaacagact
    13501 ctccgactat gatgtggatc acattgtccc tcaatctttc ctgaaggacg atagtattga
    13561 caacaaggta aagcaactgt gttttaatca atttcttgtc aggatatatg gattataact
    13621 taatttttga gaaatctgta gtatttggcg tgaaatgagt ttgctttttg gtttctcccg
    13681 tgttataggt ccttacgcgc tcagacaaga accgcggaaa atccgacaat gtacccagcg
    13741 aggaggttgt gaagaagatg aagaactatt ggaggcagct tttgaatgct aagctcataa
    13801 cccaacggaa attcgacaat ctcacgaagg cagaaagggg cggactgtct gagctcgaca
    13861 aagccggctt catcaagcgc cagttggttg aaactcgtca gattacgaaa catgtggccc
    13921 agatactcga ttcgcgtatg aatacgaagt atgatgagaa tgacaaactt atcagggagg
    13981 taaaggtaaa gtttccaact ttcctttacc atatcaaact aaagttcgaa actttttatt
    14041 tgatcaactt caaggccacc cgatctttct attcctgatt aatttgtgat gaatccatat
    14101 tgacttttga tggttacgca ggtgatcacc ctcaagagca aactggttag tgacttccgg
    14161 aaggacttcc agttttacaa ggttcgcgag atcaacaact accatcatgc ccatgacgcc
    14221 tacctgaacg ccgttgttgg cactgctctc atcaagaagt atccgaaact ggagtctgag
    14281 tttgtgtacg gggattacaa ggtgtacgac gttaggaaga tgatcgcgaa gtcagaacaa
    14341 gagatcggca aggctaccgc gaaatacttc ttttactcga atatcatgaa cttcttcaag
    14401 acagagatca ctctggcgaa tggtgaaatc cggaagaggc ctctgatcga gacaaatggc
    14461 gaaacaggtc tgtctttcct atttcatatg tttaatccta ggaatttgat caattgattg
    14521 tatgtatgtc gatcccaaga ctttcttgtt cacttatatc ttaactctct ctttgctgtt
    14581 tcttgcaggt gagattgtct gggataaggg cagggatttt gcgactgtgc gtaaggttct
    14641 cagcatgccc caagtcaaca tagtcaagaa aacggaggtt caaaccggtg gtttctccaa
    14701 ggagtccatt ctccctaagc gcaactccga caaactgatt gcgaggaaga aggattggga
    14761 tccgaagaaa tacggaggct ttgatagccc taccgtggca tacagcgtac tggtagtggc
    14821 caaggtggag aagggcaaga gcaagaaact gaaaagcgtc aaggaactgc ttggaattac
    14881 cataatggaa aggtcctcgt tcgagaagaa tccgatcgac ttcctcgagg ctaaaggtaa
    14941 aatattggat gccagacgat attctttctt ttgatttgta actttttcct gtcaaggtcg
    15001 ataaatttta ttttttttgg taaaaggtcg ataatttttt tttggagcca ttatgtaatt
    15061 ttcctaatta actgaaccaa aattatacaa accaggttac aaagaggtga agaaagacct
    15121 cattatcaaa ctgcccaagt attcgctttt cgaattggaa aatggcagaa aacgcatgct
    15181 ggcatctgcc ggagaactgc agaagggcaa cgagctggca ttgcccagta agtacgtcaa
    15241 cttcctgtac ttggcctcac actatgagaa gctgaagggg tcaccagagg acaacgagca
    15301 gaagcagttg tttgtcgagc agcacaagca ctatcttgat gagatcatag agcagatcag
    15361 cgaattttcc aagcgggtca ttcttgcaga cgctaacctc gataaggtaa ggacttctca
    15421 tgaatattag tggcagatta gtgttgttaa agtctttggt tagataatcg atgcctccta
    15481 attgtccatg ttttactggt tttctacaat taaaggtgct ttccgcgtac aacaagcaca
    15541 gagataagcc gataagggaa caagcggaaa acatcatcca cctgttcaca ctgaccaatc
    15601 tgggagcccc agcagccttt aagtacttcg ataccactat cgacagaaag cgctacacat
    15661 caaccaagga agtgttggac gctaccctta ttcaccaatc tattacaggg ctctatgaga
    15721 caaggataga tctgtcgcag ttgggtggtg actctagggc tgacccaaag aagaagcgta
    15781 aagtctgagc ttgatcctct agagtcaagc agatcgttca aacatttggc aataaagttt
    15841 cttaagattg aatcctgttg ccggtcttgc gatgattatc atataatttc tgttgaatta
    15901 cgttaagcat gtaataatta acatgtaatg catgacgtta tttatgagat gggtttttat
    15961 gattagagtc ccgcaattat acatttaata cgcgatagaa aacaaaatat agcgcgcaaa
    16021 ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta gatcgacgct actagaattc
    16081 gagctcggag gcccatttaa gttgaaaaca atcttcaaaa gtcccacatc gcttagataa
    16141 gaaaacgaag ctgagtttat atacagctag agtcgaagta gtgattgaga gaccgcagct
    16201 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
    16261 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
    16321 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
    16381 ttgcatgcct gcaggtcgac tctagaggat ccccgggtac cgagctcgaa ttcactggcc
    16441 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
    16501 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
    16561 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
    16621 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
    16681 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
    16741 ctcccggcat ccgcttacag acaagctgtg acggtctctg ttttagagct agaaatagca
    16801 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
    16861 tttcgcttta cgaattccca tggggagttc tagaatgtcg cggaacaaat tttaaaacta
    16921 aatcctaaat ttttctaatt ttgttgccaa tagtggatat gtgggccgta tagaaggaat
    16981 ctattgaagg cccaaaccca tactgacgag cccaaaggtt cgttttgcgt tttatgtttc
    17041 ggttcgatgc caacgccaca ttctgagcta ggcaaaaaac aaacgtgtct ttgaatagac
    17101 tcctctcgtt aacacatgca gcggctgcat ggtgacgcca ttaacacgtg gcctacaatt
    17161 gcatgatgtc tccattgaca cgtgacttct cgtctccttt cttaatatat ctaacaaaca
    17221 ctcctacctc ttccaaaata tatacacatc tttttgatca atctctcatt caaaatctca
    17281 ttctctctag taaacaagaa caaaaaaatg gcggatacag ctagaggaac ccatcacgat
    17341 atcatcggca gagatcagta cccgatgatg ggccgagatc gtgaccagta ccagatgtcc
    17401 ggacgaggat ctgactactc caagtctagg cagattgcta aagctgcaac tgctgtcaca
    17461 gctggtggtt ccctccttgt tctctccagc cttacccttg ttggaactgt catagctttg
    17521 actgttgcaa cacctctgct cgttatcttc agcccaatcc ttgtcccggc tctcatcaca
    17581 gttgcactcc tcatcaccgg ttttctttcc tctggagggt ttggcattgc cgctataacc
    17641 gttttctctt ggatttacaa gtaagcacac atttatcatc ttacttcata attttgtgca
    17701 atatgtgcat gcatgtgttg agccagtagc tttggatcaa tttttttggt cgaataacaa
    17761 atgtaacaat aagaaattgc aaattctagg gaacatttgg ttaactaaat acgaaatttg
    17821 acctagctag cttgaatgtg tctgtgtata tcatctatat aggtaaaatg cttggtatga
    17881 tacctattga ttgtgaatag gtacgcaacg ggagagcacc cacagggatc agacaagttg
    17941 gacagtgcaa ggatgaagtt gggaagcaaa gctcaggatc tgaaagacag agctcagtac
    18001 tacggacagc aacatactgg tggggaacat gaccgtgacc gtactcgtgg tggccagcac
    18061 actactatga gcgagctgat taaggagaac atgcacatga agctgtacat ggagggcacc
    18121 gtgaacaacc accacttcaa gtgcacatcc gagggcgaag gcaagcccta cgagggcacc
    18181 cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg
    18241 gctaccagct tcatgtacgg cagcagaacc ttcatcaacc acacccaggg catccccgac
    18301 ttctttaagc agtccttccc tgagggcttc acatgggaga gagtcaccac atacgaagat
    18361 gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct catctacaac
    18421 gtcaagatca gaggggtgaa cttcccatcc aacggccctg tgatgcagaa gaaaacactc
    18481 ggctgggagg ccaacaccga gatgctgtac cccgctgacg gcggcctgga aggcagaagc
    18541 gacatggccc tgaagctcgt gggcgggggc cacctgatct gcaacttcaa gaccacatac
    18601 agatccaaga aacccgctaa gaacctcaag atgcccggcg tctactatgt ggaccacaga
    18661 ctggaaagaa tcaaggaggc cgacaaagaa acctacgtcg agcagcacga ggtggctgtg
    18721 gccagatact gcgacctccc tagcaaactg gggcacaagt gagcttaccc cactgatgtc
    18781 atcgtcatag tccaataact ccaatgtcgg ggagttagtt tatgaggaat aaagtgttta
    18841 gaatttgatc agggggagat aataaaagcc gagtttgaat ctttttgtta taagtaatgt
    18901 ttatgtgtgt ttctatatgt tgtcaaatgg taccatgttt tttttcctct ctttttgtaa
    18961 cttgcaagtg ttgtgttgta ctttatttgg cttctttgta agttggtaac ggtggtctat
    19021 atatggaaaa ggtcttgttt tgttaaactt atgttagtta actggattcg tctttaacca
    19081 caaaaagttt tcaataagct acaaatttag acacgcaagc cgatgcagtc attagtacat
    19141 atatttattg caagtgatta catggcaacc caaacttcaa aaacagtagg ttgctccatt
    19201 tagtcgctca gagaggatgc acatgtgacc ga
//
