LOCUS       sequence-408840-       12538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nimble cloning-compatible vector for plant RNAi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12538)
  AUTHORS   Yan P, Tuo D, Shen W, Deng H, Zhou P, Gao X
  TITLE     A Nimble Cloning-compatible vector system for high-throughput gene 
            functional analysis in plants.
  JOURNAL   Plant Commun. 2023 Mar 13;4(2):100471. doi: 
            10.1016/j.xplc.2022.100471. Epub 2022 Nov 9.
  PUBMED    36352791
REFERENCE   2  (bases 1 to 12538)
  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: "2023-03-13"; volume: "4(2):100471. doi"; pages: " 
            10.1016/j.xplc.2022.100471. Epub 2022 Nov 9"
COMMENT     Sequence Label: pNC-Cam2304-RNAi
FEATURES             Location/Qualifiers
     source          1..12538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        181..283
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             314..619
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     intron          687..2283
                     /gene="pdk"
                     /number=3
                     /label=PDK intron
                     /note="modified pyruvate orthophosphate dikinase intron 
                     from Flaveria trinervia"
     CDS             1025..1708
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVVQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     VDPAY"
     CDS             complement(2347..2652)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     promoter        complement(2683..2785)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      2988..3240
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    3262..3286
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             4587..5216
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5645..6718
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      6784..6978
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(7214..7236)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    7322..7462
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     7377..7396
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7477..7494)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7648..8236)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7728..7747)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8323..9117)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    9542..9566
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    9644..9818
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(9875..10672)
                     /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="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(9986..10005)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     10596..10615
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(10735..11411)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    11602..11623
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11638..11643,11644..11661,11662..11668)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11676..11692
                     /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     11681..11703
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11700..11716
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11700..11716
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             11717..11944
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNRLA
                     AHPPFASWRNSEEARTDRPSQQLRSLNGEC"
     misc_feature    11725..11781
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(11785..11802)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(11785..11801)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11794..11816)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        12178..12523
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
ORIGIN
        1 agtggtctct gtccagtcct ggcctcgtcg gccattctcg actaagttgg cagcatcacc
       61 cgacgcactt tgcgccgaat aaatacctgt gacggaagat cacttcgcag aataaataaa
      121 tcctggtgtc cctgttgata ccgggaagcc ctgggccaac ttttggcgaa aatgagacgt
      181 tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt
      241 attttttgag ttatcgagat tttcaggagc taaggaagct aaacttttgc tgacgagaac
      301 agggactggt gaaatgcagt ttaaggttta cacctataaa agagagagcc gttatcgtct
      361 gtttgtggat gtacagagtg atattattga cacgcctggg cgacggatgg tgatccccct
      421 ggccagtgca cgtctgctgt cagataaagt ctcccgtgaa ctttacccgg tggtgcatat
      481 cggggatgaa agctggcgca tgatgaccac cgatatggcc agtgtgccgg tatccgttat
      541 cggggaagaa gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct
      601 gatgttctgg ggaatataaa tgtcaggctc ccttatacac agggccagtc tggccacttg
      661 tggtctgctg agaccacgag cccttggtaa ggaaataatt attttctttt ttccttttag
      721 tataaaatag ttaagtgatg ttaattagta tgattataat aatatagttg ttataattgt
      781 gaaaaaataa tttataaata tattgtttac ataaacaaca tagtaatgta aaaaaatatg
      841 acaagtgatg tgtaagacga agaagataaa agttgagagt aagtatatta tttttaatga
      901 atttgatcga acatgtaaga tgatatacgg ccgaagaggt tccaactttc accataatga
      961 aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag ctaaggaagc
     1021 taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa
     1081 agaacatttt gaggcatttc agtcagttgt tcaatgtacc tataaccaga ccgttcagct
     1141 ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt
     1201 tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga
     1261 cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac
     1321 tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat
     1381 atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat
     1441 tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa
     1501 cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca
     1561 aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt
     1621 ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc agggggggcg
     1681 taaacgccgc gtggatccgg cttactaaaa gccagataac agtatgcgta tttgcgcgct
     1741 gatttttgcg gtataagaat atatactgat atgtcggccc ataatagtaa ttctagctgg
     1801 tttgatgaat taaatatcaa tgataaaata ctatagtaaa aataagaata aataaattaa
     1861 aataatattt ttttatgatt aatagtttat tatataatta aatatctata ccattactaa
     1921 atattttagt ttaaaagtta ataaatattt tgttagaaat tccaatctgc ttgtaattta
     1981 tcaataaaca aaatattaaa taacaagcta aagtaacaaa taatatcaaa ctaatagaaa
     2041 cagtaatcta atgtaacaaa acataatcta atgctaatat aacaaagcgc aagatctatc
     2101 aattttatat agtattattt tcaatcaaca ttctcattaa tttctaaata atacttgtag
     2161 ttttattaac ttctaaatgg attgactatt aattaaatga attagtcgaa catgaataaa
     2221 caaggtaaca tgatagatca tgtcattgtg ttatcattga tcttacattt ggattgatta
     2281 cagttggtct ggtctcagca gaccacaagt ggccagactg gccctgtgta taagggagcc
     2341 tgacatttat attccccaga acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg
     2401 gctgagatca gccacttctt ccccgataac ggataccggc acactggcca tatcggtggt
     2461 catcatgcgc cagctttcat ccccgatatg caccaccggg taaagttcac gggagacttt
     2521 atctgacagc agacgtgcac tggccagggg gatcaccatc cgtcgcccag gcgtgtcaat
     2581 aatatcactc tgtacatcca caaacagacg ataacggctc tctcttttat aggtgtaaac
     2641 cttaaactgc atttcaccag tccctgttct cgtcagcaaa agtttagctt ccttagctcc
     2701 tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa
     2761 gttggaacct cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg cccagggctt
     2821 cccggtatca acagggacac caggatttat ttattctgcg aagtgatctt ccgtcacagg
     2881 tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac ttagtcgaga atggccgacg
     2941 aggccaggac tggacagaga ccactggtga ccagctcgaa tttccccgat cgttcaaaca
     3001 tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat
     3061 aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta
     3121 tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca
     3181 aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc
     3241 gggaattaaa ctatcagtgt ttgacaggat atattggcgg gtaaacctaa gagaaaagag
     3301 cgtttattag aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt
     3361 tgtatgtgca tgccaaccac agggttcccc tcgggatcaa agtactttga tccaacccct
     3421 ccgctgctat agtgcagtcg gcttctgacg ttcagtgcag ccgtcttctg aaaacgacat
     3481 gtcgcacaag tcctaagtta cgcgacaggc tgccgccctg cccttttcct ggcgttttct
     3541 tgtcgcgtgt tttagtcgca taaagtagaa tacttgcgac tagaaccgga gacattacgc
     3601 catgaacaag agcgccgccg ctggcctgct gggctatgcc cgcgtcagca ccgacgacca
     3661 ggacttgacc aaccaacggg ccgaactgca cgcggccggc tgcaccaagc tgttttccga
     3721 gaagatcacc ggcaccaggc gcgaccgccc ggagctggcc aggatgcttg accacctacg
     3781 ccctggcgac gttgtgacag tgaccaggct agaccgcctg gcccgcagca cccgcgacct
     3841 actggacatt gccgagcgca tccaggaggc cggcgcgggc ctgcgtagcc tggcagagcc
     3901 gtgggccgac accaccacgc cggccggccg catggtgttg accgtgttcg ccggcattgc
     3961 cgagttcgag cgttccctaa tcatcgaccg cacccggagc gggcgcgagg ccgccaaggc
     4021 gcgaggcgtg aagtttggcc cccgccctac cctcaccccg gcacagatcg cgcacgcccg
     4081 cgagctgatc gaccaggaag gccgcaccgt gaaagaggcg gctgcactgc ttggcgtgca
     4141 tcgctcgacc ctgtaccgcg cacttgagcg cagcgaggaa gtgacgccca ccgaggccag
     4201 gcggcgcggt gccttccgtg aggacgcatt gaccgaggcc gacgccctgg cggccgccga
     4261 gaatgaacgc caagaggaac aagcatgaaa ccgcaccagg acggccagga cgaaccgttt
     4321 ttcattaccg aagagatcga ggcggagatg atcgcggccg ggtacgtgtt cgagccgccc
     4381 gcgcacgtct caaccgtgcg gctgcatgaa atcctggccg gtttgtctga tgccaagctg
     4441 gcggcctggc cggcgagctt ggccgctgaa gaaaccgagc gccgccgtct aaaaaggtga
     4501 tgtgtatttg agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta
     4561 aataaacaaa tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg
     4621 ggtcaggcaa gacgaccatc gcaacccatc tagcccgcgc cctgcaactc gccggggccg
     4681 atgttctgtt agtcgattcc gatccccagg gcagtgcccg cgattgggcg gccgtgcggg
     4741 aagatcaacc gctaaccgtt gtcggcatcg accgcccgac gattgaccgc gacgtgaagg
     4801 ccatcggccg gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg gacttggctg
     4861 tgtccgcgat caaggcagcc gacttcgtgc tgattccggt gcagccaagc ccttacgaca
     4921 tatgggccac cgccgacctg gtggagctgg ttaagcagcg cattgaggtc acggatggaa
     4981 ggctacaagc ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg
     5041 ttgccgaggc gctggccggg tacgagctgc ccattcttga gtcccgtatc acgcagcgcg
     5101 tgagctaccc aggcactgcc gccgccggca caaccgttct tgaatcagaa cccgagggcg
     5161 acgctgcccg cgaggtccag gcgctggccg ctgaaattaa atcaaaactc atttgagtta
     5221 atgaggtaaa gagaaaatga gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc
     5281 acgcagcagc aaggctgcaa cgttggccag cctggcagac acgccagcca tgaagcgggt
     5341 caactttcag ttgccggcgg aggatcacac caagctgaag atgtacgcgg tacgccaagg
     5401 caagaccatt accgagctgc tatctgaata catcgcgcag ctaccagagt aaatgagcaa
     5461 atgaataaat gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac
     5521 aaccaggcac cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggccaggcg
     5581 taagcggctg ggttgcctgc cggccctgca atggcactgg aacccccaag cccgaggaat
     5641 cggcgtgagc ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga
     5701 cctggtggag aagttgaagg cggcgcaggc cgcccagcgg caacgcatcg aggcagaagc
     5761 acgccccggt gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc
     5821 gccggcagcc ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt
     5881 tttcgttccg atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc
     5941 cgttttccgt ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc
     6001 agacgggcac gtagaggttt ccgcagggcc ggcgggcatg gccagtgtgt gggattacga
     6061 cctggtactg atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa
     6121 gggagacaag cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg
     6181 gcgagccgat ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac
     6241 cacgcacgtt gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc
     6301 cgagggtgaa gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga
     6361 gtacatcgag atcgagctag ctgattggat gtaccgcgag atcacagaag gcaagaaccc
     6421 ggacgtgctg acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct
     6481 ctaccgcctg gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat
     6541 ctacgaacgc agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct
     6601 gatcgggtca aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc
     6661 gatcctagtc atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg
     6721 tacggagcag atgctagggc aaattgccct agcaggggaa aaaggtcgaa aaggtctctt
     6781 tcctgtggat agcacgtaca ttgggaaccc aaagccgtac attgggaacc ggaacccgta
     6841 cattgggaac ccaaagccgt acattgggaa ccggtcacac atgtaagtga ctgatataaa
     6901 agagaaaaaa ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac
     6961 ccgcctggcc tgtgcataac tgtctggcca gcgcacagcc gaagagctgc aaaaagcgcc
     7021 tacccttcgg tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc
     7081 tggccgctca aaaatggctg gcctacggcc aggcaatcta ccagggcgcg gacaagccgc
     7141 gccgtcgcca ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg
     7201 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     7261 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     7321 gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc
     7381 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     7441 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7501 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7561 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7621 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7681 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7741 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7801 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7861 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7921 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7981 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     8041 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     8101 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     8161 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     8221 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     8281 cgaaaactca cgttaaggga ttttggtcat gcattctagg tactaaaaca attcatccag
     8341 taaaatataa tattttattt tctcccaatc aggcttgatc cccagtaagt caaaaaatag
     8401 ctcgacatac tgttcttccc cgatatcctc cctgatcgac cggacgcaga aggcaatgtc
     8461 ataccacttg tccgccctgc cgcttctccc aagatcaata aagccactta ctttgccatc
     8521 tttcacaaag atgttgctgt ctcccaggtc gccgtgggaa aagacaagtt cctcttcggg
     8581 cttttccgtc tttaaaaaat catacagctc gcgcggatct ttaaatggag tgtcttcttc
     8641 ccagttttcg caatccacat cggccagatc gttattcagt aagtaatcca attcggctaa
     8701 gcggctgtct aagctattcg tatagggaca atccgatatg tcgatggagt gaaagagcct
     8761 gatgcactcc gcatacagct cgataatctt ttcagggctt tgttcatctt catactcttc
     8821 cgagcaaagg acgccatcgg cctcactcat gagcagattg ctccagccat catgccgttc
     8881 aaagtgcagg acctttggaa caggcagctt tccttccagc catagcatca tgtccttttc
     8941 ccgttccaca tcataggtgg tccctttata ccggctgtcc gtcattttta aatataggtt
     9001 ttcattttct cccaccagct tatatacctt agcaggagac attccttccg tatcttttac
     9061 gcagcggtat ttttcgatca gttttttcaa ttccggtgat attctcattt tagccattta
     9121 ttatttcctt cctcttttct acagtattta aagatacccc aagaagctaa ttataacaag
     9181 acgaactcca attcactgtt ccttgcattc taaaacctta aataccagaa aacagctttt
     9241 tcaaagttgt tttcaaagtt ggcgtataac atagtatcga cggagccgat tttgaaaccg
     9301 cggtgatcac aggcagcaac gctctgtcat cgttacaatc aacatgctac cctccgcgag
     9361 atcatccgtg tttcaaaccc ggcagcttag ttgccgttct tccgaatagc atcggtaaca
     9421 tgagcaaagt ctgccgcctt acaacggctc tcccgctgac gccgtcccgg actgatgggc
     9481 tgcctgtatc gagtggtgat tttgtgccga gctgccggtc ggggagctgt tggctggctg
     9541 gtggcaggat atattgtggt gtaaacaaat tgacgcttag acaacttaat aacacattgc
     9601 ggacgttttt aatgtactga attaacgccg aattaattcg ggggatctgg attttagtac
     9661 tggattttgg ttttaggaat tagaaatttt attgatagaa gtattttaca aatacaaata
     9721 catactaagg gtttcttata tgctcaacac atgagcgaaa ccctatagga accctaattc
     9781 ccttatctgg gaactactca cacattatta tggagaaact cgagcttgtc gatcgactct
     9841 agctagagga tcgatccgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat
     9901 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
     9961 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
    10021 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
    10081 tgatattcgg caagcaggca tcgccatgtg tcacgacgag atcctcgccg tcgggcatgc
    10141 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
    10201 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
    10261 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
    10321 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
    10381 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
    10441 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgg agttcattca
    10501 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgaa
    10561 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
    10621 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcccc atggtcgatc
    10681 gacagatctg cgaaagctcg agagagatag atttgtagag agagactggt gatttcagcg
    10741 tgtcctctcc aaatgaaatg aacttcctta tatagaggaa gggtcttgcg aaggatagtg
    10801 ggattgtgcg tcatccctta cgtcagtgga gatatcacat caatccactt gctttgaaga
    10861 cgtggttgga acgtcttctt tttccacgat gctcctcgtg ggtgggggtc catctttggg
    10921 accactgtcg gcagaggcat cttgaacgat agcctttcct ttatcgcaat gatggcattt
    10981 gtaggtgcca ccttcctttt ctactgtcct tttgatgaag tgacagatag ctgggcaatg
    11041 gaatccgagg aggtttcccg atattaccct ttgttgaaaa gtctcaatag ccctttggtc
    11101 ttctgagact gtatctttga tattcttgga gtagacgaga gtgtcgtgct ccaccatgtt
    11161 cacatcaatc cacttgcttt gaagacgtgg ttggaacgtc ttctttttcc acgatgctcc
    11221 tcgtgggtgg gggtccatct ttgggaccac tgtcggcaga ggcatcttga acgatagcct
    11281 ttcctttatc gcaatgatgg catttgtagg tgccaccttc cttttctact gtccttttga
    11341 tgaagtgaca gatagctggg caatggaatc cgaggaggtt tcccgatatt accctttgtt
    11401 gaaaagtctc aatagccctt tggtcttctg agactgtatc tttgatattc ttggagtaga
    11461 cgagagtgtc gtgctccacc atgttggcaa gctgctctag ccaatacgca aaccgcctct
    11521 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
    11581 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
    11641 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
    11701 aggaaacagc tatgacatga ttacgaattc gagctcggta cccggggatc ctctagagtc
    11761 gacctgcagg catgcaagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa
    11821 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
    11881 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
    11941 ctagagcagc ttgagcttgg atcagattgt cgtttcccgc cttcagttta gcttcatgga
    12001 gtcaaagatt caaatagagg acctaacaga actcgccgta aagactggcg aacagttcat
    12061 acagagtctc ttacgactca atgacaagaa gaaaatcttc gtcaacatgg tggagcacga
    12121 cacacttgtc tactccaaaa atatcaaaga tacagtctca gaagaccaaa gggcaattga
    12181 gacttttcaa caaagggtaa tatccggaaa cctcctcgga ttccattgcc cagctatctg
    12241 tcactttatt gtgaagatag tggaaaagga aggtggctcc tacaaatgcc atcattgcga
    12301 taaaggaaag gccatcgttg aagatgcctc tgccgacagt ggtcccaaag atggaccccc
    12361 acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga
    12421 ttgatgtgat atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga
    12481 cccttcctct atataaggaa gttcatttca tttggagaga acacggggga ctcttgac
//
