LOCUS       sequence-307951-       19001 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 2 cloning vector for a single guide RNA, already contains 
            Cas9, Kanamycin selection cassette for plant transformation and FAST
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 19001)
  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 19001)
  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: pAGM55297
FEATURES             Location/Qualifiers
     source          1..19001
                     /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"
     promoter        6803..6986
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             7007..7801
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7061..7080)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7671..7690
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      7827..8079
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             9781..9801
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             15534..15554
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      15581..15833
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    16019..16040
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(16055..16060,16061..16078,16079..16085)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    16093..16109
                     /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     16098..16120
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     16117..16133
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     16117..16133
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             16129..16452
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    16146..16202
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(16203..16220)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(16203..16219)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(16212..16234)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(16428..16447)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_RNA        16549..16624
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             17836..18531
                     /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 atccaaataa tctgcaccgg atctggatcg tttcgcatga ttgaacaaga
     7021 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     7081 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     7141 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactccagg acgaggcagc
     7201 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     7261 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     7321 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     7381 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     7441 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     7501 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgtatg cccgacggcg aggatctcgt
     7561 cgtgactcat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     7621 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     7681 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     7741 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     7801 agcgggtctg gatctagagt caagcagatc gttcaaacat ttggcaataa agtttcttaa
     7861 gattgaatcc tgttgccggt cttgcgatga ttatcatata atttctgttg aattacgtta
     7921 agcatgtaat aattaacatg taatgcatga cgttatttat gagatgggtt tttatgatta
     7981 gagtcccgca attatacatt taatacgcga tagaaaacaa aatatagcgc gcaaactagg
     8041 ataaattatc gcgcgcggtg tcatctatgt tactagatcg acgctgcaag aattcaagct
     8101 tggagctcaa cttttgattc gctatttgca gtgcacctgt ggcgttcatc acatcttttg
     8161 tgacactgtt tgcactggtc attgctatta caaaggacct tcctgatgtt gaaggagatc
     8221 gaaagtaagt aactgcacgc ataaccattt tctttccgct ctttggctca atccatttga
     8281 cagtcaaaga caatgtttaa ccagctccgt ttgatatatt gtctttatgt gtttgttcaa
     8341 gcatgtttag ttaatcatgc ctttgattga tcttgaatag gttccaaata tcaaccctgg
     8401 caacaaaact tggagtgaga aacattgcat tcctcggttc tggacttctg ctagtaaatt
     8461 atgtttcagc catatcacta gctttctaca tgcctcaggt gaattcatct atttccgtct
     8521 taactatttc ggttaattaa agcacgaaca ccattactgc atgtagaagc ttgataaact
     8581 atcgccacca atttattttt gttgcgatat tgttactttc ctcagtatgc agctttgaaa
     8641 agaccaaccc tcttatcctt taacaatgaa caggttttta gaggtagctt gatgattcct
     8701 gcacatgtga tcttggcttc aggcttaatt ttccaggtaa agcattatga gatactctta
     8761 tatctcttac atacttttga gataatgcac aagaacttca taactatatg ctttagtttc
     8821 tgcatttgac actgccaaat tcattaatct ctaatatctt tgttgttgat ctttggtaga
     8881 catgggtact agaaaaagca aactacacca aggtaaaata cttttgtaca aacataaact
     8941 cgttatcacg gaacatcaat ggagtgtata tctaacggag tgtagaaaca tttgattatt
     9001 gcaggaagct atctcaggat attatcggtt tatatggaat ctcttctacg cagagtatct
     9061 gttattcccc ttcctctagc tttcaatttc atggtgagga tatgcagttt tctttgtata
     9121 tcattcttct tcttctttgt agcttggagt caaaatcggt tccttcatgt acatacatca
     9181 aggatatgtc cttctgaatt tttatatctt gcaataaaaa tgcttgtacc aattgaaaca
     9241 ccagcttttt gagttctatg atcactgact tggttctaac caaaaaaaaa aaaatgttta
     9301 atttacatat ctaaaagtag gtttagggaa acctaaacag taaaatattt gtatattatt
     9361 cgaatttcac tcatcataaa aacttaaatt gcaccataaa attttgtttt actattaatg
     9421 atgtaatttg tgtaacttaa gataaaaata atattccgta agttaaccgg ctaaaaccac
     9481 gtataaacca gggaacctgt taaaccggtt ctttactgga taaagaaatg aaagcccatg
     9541 tagacagctc cattagagcc caaaccctaa atttctcatc tatataaaag gagtgacatt
     9601 agggtttttg ttcgtcctct taaagcttct cgttttctct gccgtctctc tcattcgcgc
     9661 gacgcaaacg atcttcaggt gatcttcttt ctccaaatcc tctctcataa ctctgatttc
     9721 gtacttgtgt atttgagctc acgctctgtt tctctcacca cagcaatggc ttctagccca
     9781 ccgaagaaga agcggaaggt cagctggaaa atggacaaga agtacagcat tggacttgat
     9841 attggtacga actcagttgg gtgggccgtt atcaccgatg aatacaaggt accttcgaag
     9901 aaatttaaag tgctgggcaa cacagatagg cacagcatta agaagaactt gatcggagct
     9961 ctgctctttg actctggaga aaccgcggag gcgacaaggc ttaaacgtac tgcgaggaga
    10021 aggtacactc gcaggaagaa cagaatctgt tatctccaag agatctttag caacgagatg
    10081 gcgaaggtaa ggatttttat gatatagtat gcttatgtat tttgtactga aagcatatcc
    10141 tgcttcattg ggatattact gaaagcattt aactacatgt aaactcactt gatgatcaat
    10201 aaacttgatt ttgcaggttg acgactcgtt cttccatcgc ctcgaggaat ccttcctggt
    10261 agaggaagat aagaaacacg agcgtcaccc catctttggg aatattgttg acgaagtagc
    10321 ctatcatgaa aagtatccga ctatatacca ccttcgcaag aagctggtgg actcaaccga
    10381 taaggcagac cttcggctca tatacctggc tctcgcgcac atgataaagt ttcgtggcca
    10441 tttcttgatc gaaggggacc tcaacccgga taactccgat gtggataaac tgttcattca
    10501 gctcgtccaa acctacaatc agctgttcga ggagaacccc atcaatgcat caggtaacat
    10561 tccttagtta cctttctttt ctttttccat cataagttta tagattgtac atgctttgag
    10621 atttttcttt gcaaacaatc tcaggtgtcg acgccaaggc aatactgtct gccagacttt
    10681 cgaagtccag acggcttgag aatctgatcg ctcaattgcc aggcgagaag aagaacggct
    10741 tgttcgggaa tctgattgca ctgtctctgg gcctcacccc taacttcaaa agcaactttg
    10801 acctcgccga ggacgcgaag ctgcagctgt caaaggatac atacgatgat gatctggaca
    10861 atctgctcgc ccaaataggt gctcttgaaa ttggaactct tcttttgttg tctaaaccta
    10921 tcaatttctt tgcggaaatt tatttgaagc tgtagagtta aaattgagtc ttttaaactt
    10981 ttgtaggtga tcagtatgcc gacctgttct tggctgccaa gaatctgtca gacgctatct
    11041 tgctcagtga cattctgcgg gtcaacacgg agataaccaa agcgccactt agcgcctcca
    11101 tgatcaagag gtacgacgag catcaccagg atctgaccct tctgaaggct ttggttcgcc
    11161 agcaactccc cgagaagtac aaggagattt tctttgacca atcgaagaat ggctacgcag
    11221 ggtacattga tggaggtaag ttgttactta tgattgtttt cctctctgct acatgtattt
    11281 tgttgttcat ttctgtaaga tataagaatt gagttttcct ctgatgatat tattaggtgc
    11341 aagtcaggag gaattctaca aattcatcaa gcctattctg gaaaagatgg acggtacaga
    11401 ggagctgctc gttaaattga accgcgaaga tttgcttcgg aagcagcgta ccttcgacaa
    11461 tggcagcata ccgcaccaga tccacctcgg tgagctgcat gctatcttga ggaggcaaga
    11521 ggacttctat ccgttcctga aagacaacag agagaagatt gaaaagatcc tcacgttccg
    11581 cattccctac tatgtaggtt agtatcatat gaagaaatac ctagtttcag ttgatgaatg
    11641 ctattttctg acctcagttg ttctcttttg agaattattt cttttctaat ttgcctgatt
    11701 tttctattaa ttcattaggt ccactcgcac gcgggaactc gcggtttgcg tggatgacac
    11761 gcaaatccga ggagactatc acgccttgga acttcgaaga ggtcgtggac aagggtgcga
    11821 gtgcacagtc cttcatcgaa aggatgacca acttcgataa gaatctccca aatgagaaag
    11881 tcctgcccaa gcatagtctc ctgtacgaat acttcacggt ctacaacgag ctgacgaagg
    11941 tgaaatatgt gacggagggg atgcgcaaac cggccttcct gtcaggtaaa tcctggtcca
    12001 cacttttacg ataaaaacac aagattttaa actatgaact gatcaataat cattcctaaa
    12061 agaccacact tttgttttgt ttctaaagta atttttactg ttataacagg tgagcagaag
    12121 aaggccattg tcgatctctt gttcaaaacc aatcggaagg tcactgtgaa acagcttaaa
    12181 gaggactact ttaagaagat cgaatgcttt gattctgtgg aaatcagcgg cgttgaggat
    12241 aggttcaatg cctctcttgg cacataccat gacctgttga aaatcatcaa ggacaaggac
    12301 ttccttgaca acgaggagaa cgaggacatc ctcgaggaca tcgtgctgac tctcacgctg
    12361 tttgaggaca gagaaatgat cgaggagcgc cttaagactt atgcgcatct gttcgatgac
    12421 aaggtcatga agcagttgaa gaggaggaga tatacaggta agaggtcaaa aggtttccgc
    12481 aatgatccct ctttttttgt ttctctagtt tcaagaattt gggtatatga ctaacttctg
    12541 agtgttcctt gatgcatatt tgtgatgaga caaatgtttg ttctatgttt taggttgggg
    12601 aaggctctcc aggaagctca tcaacggcat ccgcgacaag caatccggca agactatact
    12661 ggactttctc aaatccgacg gttttgcgaa tcggaacttc atgcagctta ttcacgatga
    12721 ctcactgacc ttcaaagaag atatccagaa ggcccaagtg tcaggtcagg gcgatagcct
    12781 tcacgaacac atagccaacc tggctggatc gccagctata aagaagggca tactgcagac
    12841 agtgaaggtt gtggatgagc tggtgaaggt aagttctgca tttggttatg ctccttgcat
    12901 tttaggtgtt cgtcgcactt ccatttccat gaatagctaa gatttttttt ctctgcattc
    12961 attcttcttg cctcagttct aactgtttgt ggtatttttg ttttaattat tgctacaggt
    13021 catgggccgc cataagccgg agaacatcgt catcgagatg gcgagggaaa accagacgac
    13081 tcagaaaggg cagaagaact cacgggagcg catgaagcgg atagaggaag gcatcaagga
    13141 gcttgggagt cagattctga aagagcaccc agtcgaaaat actcaactcc agaacgagaa
    13201 gctgtacctc tattacctcc agaatgggag agatatgtac gtcgaccaag agctcgacat
    13261 taacagactc tccgactatg atgtggatca cattgtccct caatctttcc tgaaggacga
    13321 tagtattgac aacaaggtaa agcaactgtg ttttaatcaa tttcttgtca ggatatatgg
    13381 attataactt aatttttgag aaatctgtag tatttggcgt gaaatgagtt tgctttttgg
    13441 tttctcccgt gttataggtc cttacgcgct cagacaagaa ccgcggaaaa tccgacaatg
    13501 tacccagcga ggaggttgtg aagaagatga agaactattg gaggcagctt ttgaatgcta
    13561 agctcataac ccaacggaaa ttcgacaatc tcacgaaggc agaaaggggc ggactgtctg
    13621 agctcgacaa agccggcttc atcaagcgcc agttggttga aactcgtcag attacgaaac
    13681 atgtggccca gatactcgat tcgcgtatga atacgaagta tgatgagaat gacaaactta
    13741 tcagggaggt aaaggtaaag tttccaactt tcctttacca tatcaaacta aagttcgaaa
    13801 ctttttattt gatcaacttc aaggccaccc gatctttcta ttcctgatta atttgtgatg
    13861 aatccatatt gacttttgat ggttacgcag gtgatcaccc tcaagagcaa actggttagt
    13921 gacttccgga aggacttcca gttttacaag gttcgcgaga tcaacaacta ccatcatgcc
    13981 catgacgcct acctgaacgc cgttgttggc actgctctca tcaagaagta tccgaaactg
    14041 gagtctgagt ttgtgtacgg ggattacaag gtgtacgacg ttaggaagat gatcgcgaag
    14101 tcagaacaag agatcggcaa ggctaccgcg aaatacttct tttactcgaa tatcatgaac
    14161 ttcttcaaga cagagatcac tctggcgaat ggtgaaatcc ggaagaggcc tctgatcgag
    14221 acaaatggcg aaacaggtct gtctttccta tttcatatgt ttaatcctag gaatttgatc
    14281 aattgattgt atgtatgtcg atcccaagac tttcttgttc acttatatct taactctctc
    14341 tttgctgttt cttgcaggtg agattgtctg ggataagggc agggattttg cgactgtgcg
    14401 taaggttctc agcatgcccc aagtcaacat agtcaagaaa acggaggttc aaaccggtgg
    14461 tttctccaag gagtccattc tccctaagcg caactccgac aaactgattg cgaggaagaa
    14521 ggattgggat ccgaagaaat acggaggctt tgatagccct accgtggcat acagcgtact
    14581 ggtagtggcc aaggtggaga agggcaagag caagaaactg aaaagcgtca aggaactgct
    14641 tggaattacc ataatggaaa ggtcctcgtt cgagaagaat ccgatcgact tcctcgaggc
    14701 taaaggtaaa atattggatg ccagacgata ttctttcttt tgatttgtaa ctttttcctg
    14761 tcaaggtcga taaattttat tttttttggt aaaaggtcga taattttttt ttggagccat
    14821 tatgtaattt tcctaattaa ctgaaccaaa attatacaaa ccaggttaca aagaggtgaa
    14881 gaaagacctc attatcaaac tgcccaagta ttcgcttttc gaattggaaa atggcagaaa
    14941 acgcatgctg gcatctgccg gagaactgca gaagggcaac gagctggcat tgcccagtaa
    15001 gtacgtcaac ttcctgtact tggcctcaca ctatgagaag ctgaaggggt caccagagga
    15061 caacgagcag aagcagttgt ttgtcgagca gcacaagcac tatcttgatg agatcataga
    15121 gcagatcagc gaattttcca agcgggtcat tcttgcagac gctaacctcg ataaggtaag
    15181 gacttctcat gaatattagt ggcagattag tgttgttaaa gtctttggtt agataatcga
    15241 tgcctcctaa ttgtccatgt tttactggtt ttctacaatt aaaggtgctt tccgcgtaca
    15301 acaagcacag agataagccg ataagggaac aagcggaaaa catcatccac ctgttcacac
    15361 tgaccaatct gggagcccca gcagccttta agtacttcga taccactatc gacagaaagc
    15421 gctacacatc aaccaaggaa gtgttggacg ctacccttat tcaccaatct attacagggc
    15481 tctatgagac aaggatagat ctgtcgcagt tgggtggtga ctctagggct gacccaaaga
    15541 agaagcgtaa agtctgagct tgatcctcta gagtcaagca gatcgttcaa acatttggca
    15601 ataaagtttc ttaagattga atcctgttgc cggtcttgcg atgattatca tataatttct
    15661 gttgaattac gttaagcatg taataattaa catgtaatgc atgacgttat ttatgagatg
    15721 ggtttttatg attagagtcc cgcaattata catttaatac gcgatagaaa acaaaatata
    15781 gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct atgttactag atcgacgcta
    15841 ctagaattcg agctcggagg cccatttaag ttgaaaacaa tcttcaaaag tcccacatcg
    15901 cttagataag aaaacgaagc tgagtttata tacagctaga gtcgaagtag tgattgagag
    15961 accgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
    16021 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
    16081 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
    16141 acgccaagct tgcatgcctg caggtcgact ctagaggatc cccgggtacc gagctcgaat
    16201 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
    16261 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
    16321 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
    16381 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
    16441 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
    16501 gcttgtctgc tcccggcatc cgcttacaga caagctgtga cggtctctgt tttagagcta
    16561 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
    16621 gtgctttttt ttcgctttac gaattcccat ggggagttct agaatgtcgc ggaacaaatt
    16681 ttaaaactaa atcctaaatt tttctaattt tgttgccaat agtggatatg tgggccgtat
    16741 agaaggaatc tattgaaggc ccaaacccat actgacgagc ccaaaggttc gttttgcgtt
    16801 ttatgtttcg gttcgatgcc aacgccacat tctgagctag gcaaaaaaca aacgtgtctt
    16861 tgaatagact cctctcgtta acacatgcag cggctgcatg gtgacgccat taacacgtgg
    16921 cctacaattg catgatgtct ccattgacac gtgacttctc gtctcctttc ttaatatatc
    16981 taacaaacac tcctacctct tccaaaatat atacacatct ttttgatcaa tctctcattc
    17041 aaaatctcat tctctctagt aaacaagaac aaaaaaatgg cggatacagc tagaggaacc
    17101 catcacgata tcatcggcag agatcagtac ccgatgatgg gccgagatcg tgaccagtac
    17161 cagatgtccg gacgaggatc tgactactcc aagtctaggc agattgctaa agctgcaact
    17221 gctgtcacag ctggtggttc cctccttgtt ctctccagcc ttacccttgt tggaactgtc
    17281 atagctttga ctgttgcaac acctctgctc gttatcttca gcccaatcct tgtcccggct
    17341 ctcatcacag ttgcactcct catcaccggt tttctttcct ctggagggtt tggcattgcc
    17401 gctataaccg ttttctcttg gatttacaag taagcacaca tttatcatct tacttcataa
    17461 ttttgtgcaa tatgtgcatg catgtgttga gccagtagct ttggatcaat ttttttggtc
    17521 gaataacaaa tgtaacaata agaaattgca aattctaggg aacatttggt taactaaata
    17581 cgaaatttga cctagctagc ttgaatgtgt ctgtgtatat catctatata ggtaaaatgc
    17641 ttggtatgat acctattgat tgtgaatagg tacgcaacgg gagagcaccc acagggatca
    17701 gacaagttgg acagtgcaag gatgaagttg ggaagcaaag ctcaggatct gaaagacaga
    17761 gctcagtact acggacagca acatactggt ggggaacatg accgtgaccg tactcgtggt
    17821 ggccagcaca ctactatgag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg
    17881 gagggcaccg tgaacaacca ccacttcaag tgcacatccg agggcgaagg caagccctac
    17941 gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc
    18001 gacatcctgg ctaccagctt catgtacggc agcagaacct tcatcaacca cacccagggc
    18061 atccccgact tctttaagca gtccttccct gagggcttca catgggagag agtcaccaca
    18121 tacgaagatg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
    18181 atctacaacg tcaagatcag aggggtgaac ttcccatcca acggccctgt gatgcagaag
    18241 aaaacactcg gctgggaggc caacaccgag atgctgtacc ccgctgacgg cggcctggaa
    18301 ggcagaagcg acatggccct gaagctcgtg ggcgggggcc acctgatctg caacttcaag
    18361 accacataca gatccaagaa acccgctaag aacctcaaga tgcccggcgt ctactatgtg
    18421 gaccacagac tggaaagaat caaggaggcc gacaaagaaa cctacgtcga gcagcacgag
    18481 gtggctgtgg ccagatactg cgacctccct agcaaactgg ggcacaagtg agcttacccc
    18541 actgatgtca tcgtcatagt ccaataactc caatgtcggg gagttagttt atgaggaata
    18601 aagtgtttag aatttgatca gggggagata ataaaagccg agtttgaatc tttttgttat
    18661 aagtaatgtt tatgtgtgtt tctatatgtt gtcaaatggt accatgtttt ttttcctctc
    18721 tttttgtaac ttgcaagtgt tgtgttgtac tttatttggc ttctttgtaa gttggtaacg
    18781 gtggtctata tatggaaaag gtcttgtttt gttaaactta tgttagttaa ctggattcgt
    18841 ctttaaccac aaaaagtttt caataagcta caaatttaga cacgcaagcc gatgcagtca
    18901 ttagtacata tatttattgc aagtgattac atggcaaccc aaacttcaaa aacagtaggt
    18961 tgctccattt agtcgctcag agaggatgca catgtgaccg a
//
