LOCUS       sequence-307941-       17006 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 17006)
  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 17006)
  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: pAGM55273
FEATURES             Location/Qualifiers
     source          1..17006
                     /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"
     CDS             340..969
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             1398..2471
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGASSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      2537..2731
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        2793..2897
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2898..3692
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      3863..4451
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4352..4371
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4587..4611
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        4776..4955
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        4808..4991
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             5012..5806
                     /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(5066..5085)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5676..5695
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      5832..6084
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             7786..7806
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             13539..13559
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      13586..13838
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    14024..14045
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(14060..14065,14066..14083,14084..14090)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14098..14114
                     /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     14103..14125
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     14122..14138
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     14122..14138
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             14134..14457
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    14151..14207
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(14208..14225)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(14208..14224)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14217..14239)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(14433..14452)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_RNA        14554..14629
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             15841..16536
                     /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 tgccagccgt gcggctgcat gaaatcctgg ccggtttgtc
      181 tgatgccaag ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg
      241 tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat
      301 gatgcgatga gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt
      361 aaccagaaag gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa
      421 ctcgccgggg ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg
      481 gcggccgtgc gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac
      541 cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg
      601 gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca
      661 agcccttacg acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag
      721 gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc
      781 atcggcggtg aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt
      841 atcacgcagc gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca
      901 gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa
      961 ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg
     1021 gccgtccgag cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag
     1081 ccatgaagcg ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg
     1141 cggtacgcca aggcaagacc attaccgagc tgctatctga atacatcgcg cagctaccag
     1201 agtaaatgag caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg
     1261 gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc
     1321 ggttggccag gcgtaagcgg ctgggttgtc tgccggccct gcaatggcac tggaaccccc
     1381 aagcccgagg aatcggcgtg agcggtcgca aaccatccgg cccggtacaa atcggcgcgg
     1441 cgctgggtga tgacctggtg gagaagttga aggccgcgca ggccgcccag cggcaacgca
     1501 tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc cgctgatcga atccgcaaag
     1561 aatcccggca accgccggca gccggtgcgt cgtcgattag gaagccgccc aagggcgacg
     1621 agcaaccaga ttttttcgtt ccgatgctct atgacgtggg cacccgcgat agtcgcagca
     1681 tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg acgagctggc gaggtgatcc
     1741 gctacgagct tccagacggg cacgtagagg tttccgcagg gccggccggc atggccagtg
     1801 tgtgggatta cgacctggta ctgatggcgg tttcccatct aaccgaatcc atgaaccgat
     1861 accgggaagg gaagggagac aagcccggcc gcgtgttccg tccacacgtt gcggacgtac
     1921 tcaagttctg ccggcgagcc gatggcggaa agcagaaaga cgacctggta gaaacctgca
     1981 ttcggttaaa caccacgcac gttgccatgc agcgtacgaa gaaggccaag aacggccgcc
     2041 tggtgacggt atccgagggt gaagccttga ttagccgcta caagatcgta aagagcgaaa
     2101 ccgggcggcc ggagtacatc gagatcgagc tagctgattg gatgtaccgc gagatcacag
     2161 aaggcaagaa cccggacgtg ctgacggttc accccgatta ctttttgatc gatcccggca
     2221 tcggccgttt tctctaccgc ctggcacgcc gcgccgcagg caaggcagaa gccagatggt
     2281 tgttcaagac gatctacgaa cgcagtggca gcgccggaga gttcaagaag ttctgtttca
     2341 ccgtgcgcaa gctgatcggg tcaaatgacc tgccggagta cgatttgaag gaggaggcgg
     2401 ggcaggctgg cccgatccta gtcatgcgct accgcaacct gatcgagggc gaagcatccg
     2461 ccggttccta atgtacggag cagatgctag ggcaaattgc cctagcaggg gaaaaaggtc
     2521 gaaaaagctt ctttcctgtg gatagcacgt acattgggaa cccaaagccg tacattggga
     2581 accggaaccc gtacattggg aacccaaagc cgtacattgg gaaccggtca cacatgtaag
     2641 tgactgatat aaaagagaaa aaaggcgatt tttccgccta aaactcttta aaacttatta
     2701 aaactcttaa aacccgcctg gcctgtgcat aactgtctgg ccagcgcaca gccgaacagc
     2761 tgcaaaaagc gcccacatca aggctccgag tgcgcggaac ccctatttgt ttatttttct
     2821 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2881 attgaaaaag gaagagtatg gctaaaatga gaatatcacc ggaattgaaa aaactgatcg
     2941 aaaaataccg ctgcgtaaaa gatacggaag gaatgtctcc tgctaaggta tataagctgg
     3001 tgggagaaaa tgaaaaccta tatttaaaaa tgacggacag ccggtataaa gggaccacct
     3061 atgatgtgga acgggaaaag gacatgatgc tatggctgga aggaaagctg cctgttccaa
     3121 aggtcctgca ctttgaacgg catgatggct ggagcaatct gctcatgagt gaggccgatg
     3181 gcgtcctttg ctcggaagag tatgaagatg aacaaagccc tgaaaagatt atcgagctgt
     3241 atgcggagtg catcaggctc tttcactcca tcgacatatc ggattgtccc tatacgaata
     3301 gcttagacag ccgcttagcc gaattggatt acttactgaa taacgatctg gccgatgtgg
     3361 attgcgaaaa ctgggaagag gacactccat ttaaagatcc gcgcgagctg tatgattttt
     3421 taaagacgga aaagcccgaa gaggaacttg tcttttccca cggcgacctg ggagacagca
     3481 acatctttgt gaaagatggc aaagtaagtg gctttattga tcttgggaga agcggcaggg
     3541 cggacaagtg gtatgacatt gccttctgcg tccggtcgct cagggaggat atcggggaag
     3601 aacagtatgt cgagctattt tttgacttac tggggatcaa gcctgattgg gagaaaataa
     3661 aatattatat tttactggat gaattgtttt agctgtcaga ccaagtttac tcatatatac
     3721 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     3781 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3841 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3901 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3961 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4021 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4081 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4141 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4201 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4261 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4321 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4381 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4441 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctgctcgga tctgttggac
     4501 cggacagtag tcatggttga tgggctgcct gtatcgagtg gtgattttgt gccgagctgc
     4561 cggtcgggga gctgttggct ggctggtggc aggatatatt gtggtgtaaa caaattgacg
     4621 cttagacaac ttaataacac attgcggacg tttttaatgt actggggttg aacactctgt
     4681 gccgaattcg gatccggaga gcggagaatt aagggagtca cgttatgacc cccgccgatg
     4741 acgcgggaca agccgtttta cgtttggaac tgacagaacc gcaacgttga aggagccact
     4801 cagccgcggg tttctggagt ttaatgagct aagcacatac gtcagaaacc attattgcgc
     4861 gttcaaaagt cgcctaaggt cactatcagc tagcaaatat ttcttgtcaa aaatgctcca
     4921 ctgacgttcc ataaattccc ctcggtatcc aattagagtc tcatattcac tctcaatcca
     4981 aataatctgc accggatctg gatcgtttcg catgattgaa caagatggat tgcacgcagg
     5041 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5101 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     5161 gaccgacctg tccggtgccc tgaatgaact ccaggacgag gcagcgcggc tatcgtggct
     5221 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5281 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5341 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5401 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5461 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5521 gttcgccagg ctcaaggcgc gtatgcccga cggcgaggat ctcgtcgtga ctcatggcga
     5581 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5641 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     5701 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     5761 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gtctggatct
     5821 agagtcaagc agatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg
     5881 ccggtcttgc gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta
     5941 acatgtaatg catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat
     6001 acatttaata cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg
     6061 cggtgtcatc tatgttacta gatcgacgct gcaagaattc aagcttggag ctcaactttt
     6121 gattcgctat ttgcagtgca cctgtggcgt tcatcacatc ttttgtgaca ctgtttgcac
     6181 tggtcattgc tattacaaag gaccttcctg atgttgaagg agatcgaaag taagtaactg
     6241 cacgcataac cattttcttt ccgctctttg gctcaatcca tttgacagtc aaagacaatg
     6301 tttaaccagc tccgtttgat atattgtctt tatgtgtttg ttcaagcatg tttagttaat
     6361 catgcctttg attgatcttg aataggttcc aaatatcaac cctggcaaca aaacttggag
     6421 tgagaaacat tgcattcctc ggttctggac ttctgctagt aaattatgtt tcagccatat
     6481 cactagcttt ctacatgcct caggtgaatt catctatttc cgtcttaact atttcggtta
     6541 attaaagcac gaacaccatt actgcatgta gaagcttgat aaactatcgc caccaattta
     6601 tttttgttgc gatattgtta ctttcctcag tatgcagctt tgaaaagacc aaccctctta
     6661 tcctttaaca atgaacaggt ttttagaggt agcttgatga ttcctgcaca tgtgatcttg
     6721 gcttcaggct taattttcca ggtaaagcat tatgagatac tcttatatct cttacatact
     6781 tttgagataa tgcacaagaa cttcataact atatgcttta gtttctgcat ttgacactgc
     6841 caaattcatt aatctctaat atctttgttg ttgatctttg gtagacatgg gtactagaaa
     6901 aagcaaacta caccaaggta aaatactttt gtacaaacat aaactcgtta tcacggaaca
     6961 tcaatggagt gtatatctaa cggagtgtag aaacatttga ttattgcagg aagctatctc
     7021 aggatattat cggtttatat ggaatctctt ctacgcagag tatctgttat tccccttcct
     7081 ctagctttca atttcatggt gaggatatgc agttttcttt gtatatcatt cttcttcttc
     7141 tttgtagctt ggagtcaaaa tcggttcctt catgtacata catcaaggat atgtccttct
     7201 gaatttttat atcttgcaat aaaaatgctt gtaccaattg aaacaccagc tttttgagtt
     7261 ctatgatcac tgacttggtt ctaaccaaaa aaaaaaaaat gtttaattta catatctaaa
     7321 agtaggttta gggaaaccta aacagtaaaa tatttgtata ttattcgaat ttcactcatc
     7381 ataaaaactt aaattgcacc ataaaatttt gttttactat taatgatgta atttgtgtaa
     7441 cttaagataa aaataatatt ccgtaagtta accggctaaa accacgtata aaccagggaa
     7501 cctgttaaac cggttcttta ctggataaag aaatgaaagc ccatgtagac agctccatta
     7561 gagcccaaac cctaaatttc tcatctatat aaaaggagtg acattagggt ttttgttcgt
     7621 cctcttaaag cttctcgttt tctctgccgt ctctctcatt cgcgcgacgc aaacgatctt
     7681 caggtgatct tctttctcca aatcctctct cataactctg atttcgtact tgtgtatttg
     7741 agctcacgct ctgtttctct caccacagca atggcttcta gcccaccgaa gaagaagcgg
     7801 aaggtcagct ggaaaatgga caagaagtac agcattggac ttgatattgg tacgaactca
     7861 gttgggtggg ccgttatcac cgatgaatac aaggtacctt cgaagaaatt taaagtgctg
     7921 ggcaacacag ataggcacag cattaagaag aacttgatcg gagctctgct ctttgactct
     7981 ggagaaaccg cggaggcgac aaggcttaaa cgtactgcga ggagaaggta cactcgcagg
     8041 aagaacagaa tctgttatct ccaagagatc tttagcaacg agatggcgaa ggtaaggatt
     8101 tttatgatat agtatgctta tgtattttgt actgaaagca tatcctgctt cattgggata
     8161 ttactgaaag catttaacta catgtaaact cacttgatga tcaataaact tgattttgca
     8221 ggttgacgac tcgttcttcc atcgcctcga ggaatccttc ctggtagagg aagataagaa
     8281 acacgagcgt caccccatct ttgggaatat tgttgacgaa gtagcctatc atgaaaagta
     8341 tccgactata taccaccttc gcaagaagct ggtggactca accgataagg cagaccttcg
     8401 gctcatatac ctggctctcg cgcacatgat aaagtttcgt ggccatttct tgatcgaagg
     8461 ggacctcaac ccggataact ccgatgtgga taaactgttc attcagctcg tccaaaccta
     8521 caatcagctg ttcgaggaga accccatcaa tgcatcaggt aacattcctt agttaccttt
     8581 cttttctttt tccatcataa gtttatagat tgtacatgct ttgagatttt tctttgcaaa
     8641 caatctcagg tgtcgacgcc aaggcaatac tgtctgccag actttcgaag tccagacggc
     8701 ttgagaatct gatcgctcaa ttgccaggcg agaagaagaa cggcttgttc gggaatctga
     8761 ttgcactgtc tctgggcctc acccctaact tcaaaagcaa ctttgacctc gccgaggacg
     8821 cgaagctgca gctgtcaaag gatacatacg atgatgatct ggacaatctg ctcgcccaaa
     8881 taggtgctct tgaaattgga actcttcttt tgttgtctaa acctatcaat ttctttgcgg
     8941 aaatttattt gaagctgtag agttaaaatt gagtctttta aacttttgta ggtgatcagt
     9001 atgccgacct gttcttggct gccaagaatc tgtcagacgc tatcttgctc agtgacattc
     9061 tgcgggtcaa cacggagata accaaagcgc cacttagcgc ctccatgatc aagaggtacg
     9121 acgagcatca ccaggatctg acccttctga aggctttggt tcgccagcaa ctccccgaga
     9181 agtacaagga gattttcttt gaccaatcga agaatggcta cgcagggtac attgatggag
     9241 gtaagttgtt acttatgatt gttttcctct ctgctacatg tattttgttg ttcatttctg
     9301 taagatataa gaattgagtt ttcctctgat gatattatta ggtgcaagtc aggaggaatt
     9361 ctacaaattc atcaagccta ttctggaaaa gatggacggt acagaggagc tgctcgttaa
     9421 attgaaccgc gaagatttgc ttcggaagca gcgtaccttc gacaatggca gcataccgca
     9481 ccagatccac ctcggtgagc tgcatgctat cttgaggagg caagaggact tctatccgtt
     9541 cctgaaagac aacagagaga agattgaaaa gatcctcacg ttccgcattc cctactatgt
     9601 aggttagtat catatgaaga aatacctagt ttcagttgat gaatgctatt ttctgacctc
     9661 agttgttctc ttttgagaat tatttctttt ctaatttgcc tgatttttct attaattcat
     9721 taggtccact cgcacgcggg aactcgcggt ttgcgtggat gacacgcaaa tccgaggaga
     9781 ctatcacgcc ttggaacttc gaagaggtcg tggacaaggg tgcgagtgca cagtccttca
     9841 tcgaaaggat gaccaacttc gataagaatc tcccaaatga gaaagtcctg cccaagcata
     9901 gtctcctgta cgaatacttc acggtctaca acgagctgac gaaggtgaaa tatgtgacgg
     9961 aggggatgcg caaaccggcc ttcctgtcag gtaaatcctg gtccacactt ttacgataaa
    10021 aacacaagat tttaaactat gaactgatca ataatcattc ctaaaagacc acacttttgt
    10081 tttgtttcta aagtaatttt tactgttata acaggtgagc agaagaaggc cattgtcgat
    10141 ctcttgttca aaaccaatcg gaaggtcact gtgaaacagc ttaaagagga ctactttaag
    10201 aagatcgaat gctttgattc tgtggaaatc agcggcgttg aggataggtt caatgcctct
    10261 cttggcacat accatgacct gttgaaaatc atcaaggaca aggacttcct tgacaacgag
    10321 gagaacgagg acatcctcga ggacatcgtg ctgactctca cgctgtttga ggacagagaa
    10381 atgatcgagg agcgccttaa gacttatgcg catctgttcg atgacaaggt catgaagcag
    10441 ttgaagagga ggagatatac aggtaagagg tcaaaaggtt tccgcaatga tccctctttt
    10501 tttgtttctc tagtttcaag aatttgggta tatgactaac ttctgagtgt tccttgatgc
    10561 atatttgtga tgagacaaat gtttgttcta tgttttaggt tggggaaggc tctccaggaa
    10621 gctcatcaac ggcatccgcg acaagcaatc cggcaagact atactggact ttctcaaatc
    10681 cgacggtttt gcgaatcgga acttcatgca gcttattcac gatgactcac tgaccttcaa
    10741 agaagatatc cagaaggccc aagtgtcagg tcagggcgat agccttcacg aacacatagc
    10801 caacctggct ggatcgccag ctataaagaa gggcatactg cagacagtga aggttgtgga
    10861 tgagctggtg aaggtaagtt ctgcatttgg ttatgctcct tgcattttag gtgttcgtcg
    10921 cacttccatt tccatgaata gctaagattt tttttctctg cattcattct tcttgcctca
    10981 gttctaactg tttgtggtat ttttgtttta attattgcta caggtcatgg gccgccataa
    11041 gccggagaac atcgtcatcg agatggcgag ggaaaaccag acgactcaga aagggcagaa
    11101 gaactcacgg gagcgcatga agcggataga ggaaggcatc aaggagcttg ggagtcagat
    11161 tctgaaagag cacccagtcg aaaatactca actccagaac gagaagctgt acctctatta
    11221 cctccagaat gggagagata tgtacgtcga ccaagagctc gacattaaca gactctccga
    11281 ctatgatgtg gatcacattg tccctcaatc tttcctgaag gacgatagta ttgacaacaa
    11341 ggtaaagcaa ctgtgtttta atcaatttct tgtcaggata tatggattat aacttaattt
    11401 ttgagaaatc tgtagtattt ggcgtgaaat gagtttgctt tttggtttct cccgtgttat
    11461 aggtccttac gcgctcagac aagaaccgcg gaaaatccga caatgtaccc agcgaggagg
    11521 ttgtgaagaa gatgaagaac tattggaggc agcttttgaa tgctaagctc ataacccaac
    11581 ggaaattcga caatctcacg aaggcagaaa ggggcggact gtctgagctc gacaaagccg
    11641 gcttcatcaa gcgccagttg gttgaaactc gtcagattac gaaacatgtg gcccagatac
    11701 tcgattcgcg tatgaatacg aagtatgatg agaatgacaa acttatcagg gaggtaaagg
    11761 taaagtttcc aactttcctt taccatatca aactaaagtt cgaaactttt tatttgatca
    11821 acttcaaggc cacccgatct ttctattcct gattaatttg tgatgaatcc atattgactt
    11881 ttgatggtta cgcaggtgat caccctcaag agcaaactgg ttagtgactt ccggaaggac
    11941 ttccagtttt acaaggttcg cgagatcaac aactaccatc atgcccatga cgcctacctg
    12001 aacgccgttg ttggcactgc tctcatcaag aagtatccga aactggagtc tgagtttgtg
    12061 tacggggatt acaaggtgta cgacgttagg aagatgatcg cgaagtcaga acaagagatc
    12121 ggcaaggcta ccgcgaaata cttcttttac tcgaatatca tgaacttctt caagacagag
    12181 atcactctgg cgaatggtga aatccggaag aggcctctga tcgagacaaa tggcgaaaca
    12241 ggtctgtctt tcctatttca tatgtttaat cctaggaatt tgatcaattg attgtatgta
    12301 tgtcgatccc aagactttct tgttcactta tatcttaact ctctctttgc tgtttcttgc
    12361 aggtgagatt gtctgggata agggcaggga ttttgcgact gtgcgtaagg ttctcagcat
    12421 gccccaagtc aacatagtca agaaaacgga ggttcaaacc ggtggtttct ccaaggagtc
    12481 cattctccct aagcgcaact ccgacaaact gattgcgagg aagaaggatt gggatccgaa
    12541 gaaatacgga ggctttgata gccctaccgt ggcatacagc gtactggtag tggccaaggt
    12601 ggagaagggc aagagcaaga aactgaaaag cgtcaaggaa ctgcttggaa ttaccataat
    12661 ggaaaggtcc tcgttcgaga agaatccgat cgacttcctc gaggctaaag gtaaaatatt
    12721 ggatgccaga cgatattctt tcttttgatt tgtaactttt tcctgtcaag gtcgataaat
    12781 tttatttttt ttggtaaaag gtcgataatt tttttttgga gccattatgt aattttccta
    12841 attaactgaa ccaaaattat acaaaccagg ttacaaagag gtgaagaaag acctcattat
    12901 caaactgccc aagtattcgc ttttcgaatt ggaaaatggc agaaaacgca tgctggcatc
    12961 tgccggagaa ctgcagaagg gcaacgagct ggcattgccc agtaagtacg tcaacttcct
    13021 gtacttggcc tcacactatg agaagctgaa ggggtcacca gaggacaacg agcagaagca
    13081 gttgtttgtc gagcagcaca agcactatct tgatgagatc atagagcaga tcagcgaatt
    13141 ttccaagcgg gtcattcttg cagacgctaa cctcgataag gtaaggactt ctcatgaata
    13201 ttagtggcag attagtgttg ttaaagtctt tggttagata atcgatgcct cctaattgtc
    13261 catgttttac tggttttcta caattaaagg tgctttccgc gtacaacaag cacagagata
    13321 agccgataag ggaacaagcg gaaaacatca tccacctgtt cacactgacc aatctgggag
    13381 ccccagcagc ctttaagtac ttcgatacca ctatcgacag aaagcgctac acatcaacca
    13441 aggaagtgtt ggacgctacc cttattcacc aatctattac agggctctat gagacaagga
    13501 tagatctgtc gcagttgggt ggtgactcta gggctgaccc aaagaagaag cgtaaagtct
    13561 gagcttgatc ctctagagtc aagcagatcg ttcaaacatt tggcaataaa gtttcttaag
    13621 attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga attacgttaa
    13681 gcatgtaata attaacatgt aatgcatgac gttatttatg agatgggttt ttatgattag
    13741 agtcccgcaa ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga
    13801 taaattatcg cgcgcggtgt catctatgtt actagatcga cgctactaga attcgagctc
    13861 ggaggcccat ttaagttgaa aacaatcttc aaaagtccca catcgcttag ataagaaaac
    13921 gaagctgagt ttatatacag ctagagtcga agtagtgatt gagagaccgc agctggcacg
    13981 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
    14041 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
    14101 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcttgcat
    14161 gcctgcaggt cgactctaga ggatccccgg gtaccgagct cgaattcact ggccgtcgtt
    14221 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
    14281 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
    14341 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
    14401 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
    14461 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
    14521 gcatccgctt acagacaagc tgtgacggtc tctgttttag agctagaaat agcaagttaa
    14581 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttttcgc
    14641 tttacgaatt cccatgggga gttctagaat gtcgcggaac aaattttaaa actaaatcct
    14701 aaatttttct aattttgttg ccaatagtgg atatgtgggc cgtatagaag gaatctattg
    14761 aaggcccaaa cccatactga cgagcccaaa ggttcgtttt gcgttttatg tttcggttcg
    14821 atgccaacgc cacattctga gctaggcaaa aaacaaacgt gtctttgaat agactcctct
    14881 cgttaacaca tgcagcggct gcatggtgac gccattaaca cgtggcctac aattgcatga
    14941 tgtctccatt gacacgtgac ttctcgtctc ctttcttaat atatctaaca aacactccta
    15001 cctcttccaa aatatataca catctttttg atcaatctct cattcaaaat ctcattctct
    15061 ctagtaaaca agaacaaaaa aatggcggat acagctagag gaacccatca cgatatcatc
    15121 ggcagagatc agtacccgat gatgggccga gatcgtgacc agtaccagat gtccggacga
    15181 ggatctgact actccaagtc taggcagatt gctaaagctg caactgctgt cacagctggt
    15241 ggttccctcc ttgttctctc cagccttacc cttgttggaa ctgtcatagc tttgactgtt
    15301 gcaacacctc tgctcgttat cttcagccca atccttgtcc cggctctcat cacagttgca
    15361 ctcctcatca ccggttttct ttcctctgga gggtttggca ttgccgctat aaccgttttc
    15421 tcttggattt acaagtaagc acacatttat catcttactt cataattttg tgcaatatgt
    15481 gcatgcatgt gttgagccag tagctttgga tcaatttttt tggtcgaata acaaatgtaa
    15541 caataagaaa ttgcaaattc tagggaacat ttggttaact aaatacgaaa tttgacctag
    15601 ctagcttgaa tgtgtctgtg tatatcatct atataggtaa aatgcttggt atgataccta
    15661 ttgattgtga ataggtacgc aacgggagag cacccacagg gatcagacaa gttggacagt
    15721 gcaaggatga agttgggaag caaagctcag gatctgaaag acagagctca gtactacgga
    15781 cagcaacata ctggtgggga acatgaccgt gaccgtactc gtggtggcca gcacactact
    15841 atgagcgagc tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtgaac
    15901 aaccaccact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc
    15961 atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctacc
    16021 agcttcatgt acggcagcag aaccttcatc aaccacaccc agggcatccc cgacttcttt
    16081 aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga agatgggggc
    16141 gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag
    16201 atcagagggg tgaacttccc atccaacggc cctgtgatgc agaagaaaac actcggctgg
    16261 gaggccaaca ccgagatgct gtaccccgct gacggcggcc tggaaggcag aagcgacatg
    16321 gccctgaagc tcgtgggcgg gggccacctg atctgcaact tcaagaccac atacagatcc
    16381 aagaaacccg ctaagaacct caagatgccc ggcgtctact atgtggacca cagactggaa
    16441 agaatcaagg aggccgacaa agaaacctac gtcgagcagc acgaggtggc tgtggccaga
    16501 tactgcgacc tccctagcaa actggggcac aagtgagctt accccactga tgtcatcgtc
    16561 atagtccaat aactccaatg tcggggagtt agtttatgag gaataaagtg tttagaattt
    16621 gatcaggggg agataataaa agccgagttt gaatcttttt gttataagta atgtttatgt
    16681 gtgtttctat atgttgtcaa atggtaccat gttttttttc ctctcttttt gtaacttgca
    16741 agtgttgtgt tgtactttat ttggcttctt tgtaagttgg taacggtggt ctatatatgg
    16801 aaaaggtctt gttttgttaa acttatgtta gttaactgga ttcgtcttta accacaaaaa
    16861 gttttcaata agctacaaat ttagacacgc aagccgatgc agtcattagt acatatattt
    16921 attgcaagtg attacatggc aacccaaact tcaaaaacag taggttgctc catttagtcg
    16981 ctcagagagg atgcacatgt gaccga
//
