LOCUS       sequence-379473-       13004 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nimble cloning-compatible vector for plant ectopic expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13004)
  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 13004)
  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-Cam1304-MCS
FEATURES             Location/Qualifiers
     source          1..13004
                     /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"
     primer_bind     498..517
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(693..710)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(693..709)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(702..724)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        1086..1431
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             1463..2173
                     /codon_start=1
                     /gene="mgfp5"
                     /product="GFP with folding enhancement mutations"
                     /label=mgfp5
                     /note="suitable for expression in plants due to removal of 
                     a cryptic intron"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKAN
                     FKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1490..1518)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2113..2133
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2180..3985
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /translation="LRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAIA
                     VPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEVM
                     EHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHDF
                     FNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQV
                     VATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQFL
                     INHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDWA
                     DEHGIVVIDETAAVGFQLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARDK
                     NHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISDL
                     FDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMYT
                     DMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRKP
                     KSAAFLLQKRWTGMNFGEKPQQGGKQ"
     CDS             3992..4009
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4041..4293
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    4315..4339
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             5640..6269
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             6698..7771
                     /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      7837..8031
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(8267..8289)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    8375..8515
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     8430..8449
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(8530..8547)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8701..9289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8781..8800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9376..10170)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    10595..10619
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    10697..10871
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(10912..11937)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     promoter        complement(12004..12680)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    12871..12892
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12907..12912,12913..12930,12931..12937)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12945..12961
                     /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     12950..12972
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12969..12985
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12969..12985
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
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 agaccaggca tgcaagcttg gcactggccg tcgttttaca acgtcgtgac
      721 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
      781 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
      841 ggcgaatgct agagcagctt gagcttggat cagattgtcg tttcccgcct tcagtttagc
      901 ttcatggagt caaagattca aatagaggac ctaacagaac tcgccgtaaa gactggcgaa
      961 cagttcatac agagtctctt acgactcaat gacaagaaga aaatcttcgt caacatggtg
     1021 gagcacgaca cacttgtcta ctccaaaaat atcaaagata cagtctcaga agaccaaagg
     1081 gcaattgaga cttttcaaca aagggtaata tccggaaacc tcctcggatt ccattgccca
     1141 gctatctgtc actttattgt gaagatagtg gaaaaggaag gtggctccta caaatgccat
     1201 cattgcgata aaggaaaggc catcgttgaa gatgcctctg ccgacagtgg tcccaaagat
     1261 ggacccccac ccacgaggag catcgtggaa aaagaagacg ttccaaccac gtcttcaaag
     1321 caagtggatt gatgtgatat ctccactgac gtaagggatg acgcacaatc ccactatcct
     1381 tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagagaac acgggggact
     1441 cttgaccatg gtagatctga ctagtaaagg agaagaactt ttcactggag ttgtcccaat
     1501 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga
     1561 aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc
     1621 tgttccgtgg ccaacacttg tcactacttt ctcttatggt gttcaatgct tttcaagata
     1681 cccagatcat atgaagcggc acgacttctt caagagcgcc atgcctgagg gatacgtgca
     1741 ggagaggacc atcttcttca aggacgacgg gaactacaag acacgtgctg aagtcaagtt
     1801 tgagggagac accctcgtca acaggatcga gcttaaggga atcgatttca aggaggacgg
     1861 aaacatcctc ggccacaagt tggaatacaa ctacaactcc cacaacgtat acatcatggc
     1921 cgacaagcaa aagaacggca tcaaagccaa cttcaagacc cgccacaaca tcgaagacgg
     1981 cggcgtgcaa ctcgctgatc attatcaaca aaatactcca attggcgatg gccctgtcct
     2041 tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa
     2101 gagagaccac atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga
     2161 tgaactatac aaagctagtt tacgtcctgt agaaacccca acccgtgaaa tcaaaaaact
     2221 cgacggcctg tgggcattca gtctggatcg cgaaaactgt ggaattgatc agcgttggtg
     2281 ggaaagcgcg ttacaagaaa gccgggcaat tgctgtgcca ggcagtttta acgatcagtt
     2341 cgccgatgca gatattcgta attatgcggg caacgtctgg tatcagcgcg aagtctttat
     2401 accgaaaggt tgggcaggcc agcgtatcgt gctgcgtttc gatgcggtca ctcattacgg
     2461 caaagtgtgg gtcaataatc aggaagtgat ggagcatcag ggcggctata cgccatttga
     2521 agccgatgtc acgccgtatg ttattgccgg gaaaagtgta cgtatcaccg tttgtgtgaa
     2581 caacgaactg aactggcaga ctatcccgcc gggaatggtg attaccgacg aaaacggcaa
     2641 gaaaaagcag tcttacttcc atgatttctt taactatgcc ggaatccatc gcagcgtaat
     2701 gctctacacc acgccgaaca cctgggtgga cgatatcacc gtggtgacgc atgtcgcgca
     2761 agactgtaac cacgcgtctg ttgactggca ggtggtggcc aatggtgatg tcagcgttga
     2821 actgcgtgat gcggatcaac aggtggttgc aactggacaa ggcactagcg ggactttgca
     2881 agtggtgaat ccgcacctct ggcaaccggg tgaaggttat ctctatgaac tgtgcgtcac
     2941 agccaaaagc cagacagagt gtgatatcta cccgcttcgc gtcggcatcc ggtcagtggc
     3001 agtgaagggc gaacagttcc tgattaacca caaaccgttc tactttactg gctttggtcg
     3061 tcatgaagat gcggacttgc gtggcaaagg attcgataac gtgctgatgg tgcacgacca
     3121 cgcattaatg gactggattg gggccaactc ctaccgtacc tcgcattacc cttacgctga
     3181 agagatgctc gactgggcag atgaacatgg catcgtggtg attgatgaaa ctgctgctgt
     3241 cggctttcag ctgtctttag gcattggttt cgaagcgggc aacaagccga aagaactgta
     3301 cagcgaagag gcagtcaacg gggaaactca gcaagcgcac ttacaggcga ttaaagagct
     3361 gatagcgcgt gacaaaaacc acccaagcgt ggtgatgtgg agtattgcca acgaaccgga
     3421 tacccgtccg caaggtgcac gggaatattt cgcgccactg gcggaagcaa cgcgtaaact
     3481 cgacccgacg cgtccgatca cctgcgtcaa tgtaatgttc tgcgacgctc acaccgatac
     3541 catcagcgat ctctttgatg tgctgtgcct gaaccgttat tacggatggt atgtccaaag
     3601 cggcgatttg gaaacggcag agaaggtact ggaaaaagaa cttctggcct ggcaggagaa
     3661 actgcatcag ccgattatca tcaccgaata cggcgtggat acgttagccg ggctgcactc
     3721 aatgtacacc gacatgtgga gtgaagagta tcagtgtgca tggctggata tgtatcaccg
     3781 cgtctttgat cgcgtcagcg ccgtcgtcgg tgaacaggta tggaatttcg ccgattttgc
     3841 gacctcgcaa ggcatattgc gcgttggcgg taacaagaaa gggatcttca ctcgcgaccg
     3901 caaaccgaag tcggcggctt ttctgctgca aaaacgctgg actggcatga acttcggtga
     3961 aaaaccgcag cagggaggca aacaagctag ccaccaccac caccaccacg tgtgaattgg
     4021 tgaccagctc gaatttcccc gatcgttcaa acatttggca ataaagtttc ttaagattga
     4081 atcctgttgc cggtcttgcg atgattatca tataatttct gttgaattac gttaagcatg
     4141 taataattaa catgtaatgc atgacgttat ttatgagatg ggtttttatg attagagtcc
     4201 cgcaattata catttaatac gcgatagaaa acaaaatata gcgcgcaaac taggataaat
     4261 tatcgcgcgc ggtgtcatct atgttactag atcgggaatt aaactatcag tgtttgacag
     4321 gatatattgg cgggtaaacc taagagaaaa gagcgtttat tagaataatc ggatatttaa
     4381 aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt gcatgccaac cacagggttc
     4441 ccctcgggat caaagtactt tgatccaacc cctccgctgc tatagtgcag tcggcttctg
     4501 acgttcagtg cagccgtctt ctgaaaacga catgtcgcac aagtcctaag ttacgcgaca
     4561 ggctgccgcc ctgccctttt cctggcgttt tcttgtcgcg tgttttagtc gcataaagta
     4621 gaatacttgc gactagaacc ggagacatta cgccatgaac aagagcgccg ccgctggcct
     4681 gctgggctat gcccgcgtca gcaccgacga ccaggacttg accaaccaac gggccgaact
     4741 gcacgcggcc ggctgcacca agctgttttc cgagaagatc accggcacca ggcgcgaccg
     4801 cccggagctg gccaggatgc ttgaccacct acgccctggc gacgttgtga cagtgaccag
     4861 gctagaccgc ctggcccgca gcacccgcga cctactggac attgccgagc gcatccagga
     4921 ggccggcgcg ggcctgcgta gcctggcaga gccgtgggcc gacaccacca cgccggccgg
     4981 ccgcatggtg ttgaccgtgt tcgccggcat tgccgagttc gagcgttccc taatcatcga
     5041 ccgcacccgg agcgggcgcg aggccgccaa ggcgcgaggc gtgaagtttg gcccccgccc
     5101 taccctcacc ccggcacaga tcgcgcacgc ccgcgagctg atcgaccagg aaggccgcac
     5161 cgtgaaagag gcggctgcac tgcttggcgt gcatcgctcg accctgtacc gcgcacttga
     5221 gcgcagcgag gaagtgacgc ccaccgaggc caggcggcgc ggtgccttcc gtgaggacgc
     5281 attgaccgag gccgacgccc tggcggccgc cgagaatgaa cgccaagagg aacaagcatg
     5341 aaaccgcacc aggacggcca ggacgaaccg tttttcatta ccgaagagat cgaggcggag
     5401 atgatcgcgg ccgggtacgt gttcgagccg cccgcgcacg tctcaaccgt gcggctgcat
     5461 gaaatcctgg ccggtttgtc tgatgccaag ctggcggcct ggccggcgag cttggccgct
     5521 gaagaaaccg agcgccgccg tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt
     5581 catgcggtcg ctgcgtatat gatgcgatga gtaaataaac aaatacgcaa ggggaacgca
     5641 tgaaggttat cgctgtactt aaccagaaag gcgggtcagg caagacgacc atcgcaaccc
     5701 atctagcccg cgccctgcaa ctcgccgggg ccgatgttct gttagtcgat tccgatcccc
     5761 agggcagtgc ccgcgattgg gcggccgtgc gggaagatca accgctaacc gttgtcggca
     5821 tcgaccgccc gacgattgac cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga
     5881 tcgacggagc gccccaggcg gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg
     5941 tgctgattcc ggtgcagcca agcccttacg acatatgggc caccgccgac ctggtggagc
     6001 tggttaagca gcgcattgag gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc
     6061 gggcgatcaa aggcacgcgc atcggcggtg aggttgccga ggcgctggcc gggtacgagc
     6121 tgcccattct tgagtcccgt atcacgcagc gcgtgagcta cccaggcact gccgccgccg
     6181 gcacaaccgt tcttgaatca gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg
     6241 ccgctgaaat taaatcaaaa ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag
     6301 cacaaacacg ctaagtgccg gccgtccgag cgcacgcagc agcaaggctg caacgttggc
     6361 cagcctggca gacacgccag ccatgaagcg ggtcaacttt cagttgccgg cggaggatca
     6421 caccaagctg aagatgtacg cggtacgcca aggcaagacc attaccgagc tgctatctga
     6481 atacatcgcg cagctaccag agtaaatgag caaatgaata aatgagtaga tgaattttag
     6541 cggctaaagg aggcggcatg gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc
     6601 ccatgtgtgg aggaacgggc ggttggccag gcgtaagcgg ctgggttgcc tgccggccct
     6661 gcaatggcac tggaaccccc aagcccgagg aatcggcgtg agcggtcgca aaccatccgg
     6721 cccggtacaa atcggcgcgg cgctgggtga tgacctggtg gagaagttga aggcggcgca
     6781 ggccgcccag cggcaacgca tcgaggcaga agcacgcccc ggtgaatcgt ggcaagcggc
     6841 cgctgatcga atccgcaaag aatcccggca accgccggca gccggtgcgc cgtcgattag
     6901 gaagccgccc aagggcgacg agcaaccaga ttttttcgtt ccgatgctct atgacgtggg
     6961 cacccgcgat agtcgcagca tcatggacgt ggccgttttc cgtctgtcga agcgtgaccg
     7021 acgagctggc gaggtgatcc gctacgagct tccagacggg cacgtagagg tttccgcagg
     7081 gccggcgggc atggccagtg tgtgggatta cgacctggta ctgatggcgg tttcccatct
     7141 aaccgaatcc atgaaccgat accgggaagg gaagggagac aagcccggcc gcgtgttccg
     7201 tccacacgtt gcggacgtac tcaagttctg ccggcgagcc gatggcggaa agcagaaaga
     7261 cgacctggta gaaacctgca ttcggttaaa caccacgcac gttgccatgc agcgtacgaa
     7321 gaaggccaag aacggccgcc tggtgacggt atccgagggt gaagccttga ttagccgcta
     7381 caagatcgta aagagcgaaa ccgggcggcc ggagtacatc gagatcgagc tagctgattg
     7441 gatgtaccgc gagatcacag aaggcaagaa cccggacgtg ctgacggttc accccgatta
     7501 ctttttgatc gatcccggca tcggccgttt tctctaccgc ctggcacgcc gcgccgcagg
     7561 caaggcagaa gccagatggt tgttcaagac gatctacgaa cgcagtggca gcgccggaga
     7621 gttcaagaag ttctgtttca ccgtgcgcaa gctgatcggg tcaaatgacc tgccggagta
     7681 cgatttgaag gaggaggcgg ggcaggctgg cccgatccta gtcatgcgct accgcaacct
     7741 gatcgagggc gaagcatccg ccggttccta atgtacggag cagatgctag ggcaaattgc
     7801 cctagcaggg gaaaaaggtc gaaaaggtct ctttcctgtg gatagcacgt acattgggaa
     7861 cccaaagccg tacattggga accggaaccc gtacattggg aacccaaagc cgtacattgg
     7921 gaaccggtca cacatgtaag tgactgatat aaaagagaaa aaaggcgatt tttccgccta
     7981 aaactcttta aaacttatta aaactcttaa aacccgcctg gcctgtgcat aactgtctgg
     8041 ccagcgcaca gccgaagagc tgcaaaaagc gcctaccctt cggtcgctgc gctccctacg
     8101 ccccgccgct tcgcgtcggc ctatcgcggc cgctggccgc tcaaaaatgg ctggcctacg
     8161 gccaggcaat ctaccagggc gcggacaagc cgcgccgtcg ccactcgacc gccggcgccc
     8221 acatcaaggc accctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     8281 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     8341 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
     8401 atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     8461 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc
     8521 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     8581 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     8641 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     8701 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     8761 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     8821 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     8881 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     8941 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     9001 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     9061 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     9121 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     9181 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     9241 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     9301 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     9361 catgcattct aggtactaaa acaattcatc cagtaaaata taatatttta ttttctccca
     9421 atcaggcttg atccccagta agtcaaaaaa tagctcgaca tactgttctt ccccgatatc
     9481 ctccctgatc gaccggacgc agaaggcaat gtcataccac ttgtccgccc tgccgcttct
     9541 cccaagatca ataaagccac ttactttgcc atctttcaca aagatgttgc tgtctcccag
     9601 gtcgccgtgg gaaaagacaa gttcctcttc gggcttttcc gtctttaaaa aatcatacag
     9661 ctcgcgcgga tctttaaatg gagtgtcttc ttcccagttt tcgcaatcca catcggccag
     9721 atcgttattc agtaagtaat ccaattcggc taagcggctg tctaagctat tcgtataggg
     9781 acaatccgat atgtcgatgg agtgaaagag cctgatgcac tccgcataca gctcgataat
     9841 cttttcaggg ctttgttcat cttcatactc ttccgagcaa aggacgccat cggcctcact
     9901 catgagcaga ttgctccagc catcatgccg ttcaaagtgc aggacctttg gaacaggcag
     9961 ctttccttcc agccatagca tcatgtcctt ttcccgttcc acatcatagg tggtcccttt
    10021 ataccggctg tccgtcattt ttaaatatag gttttcattt tctcccacca gcttatatac
    10081 cttagcagga gacattcctt ccgtatcttt tacgcagcgg tatttttcga tcagtttttt
    10141 caattccggt gatattctca ttttagccat ttattatttc cttcctcttt tctacagtat
    10201 ttaaagatac cccaagaagc taattataac aagacgaact ccaattcact gttccttgca
    10261 ttctaaaacc ttaaatacca gaaaacagct ttttcaaagt tgttttcaaa gttggcgtat
    10321 aacatagtat cgacggagcc gattttgaaa ccgcggtgat cacaggcagc aacgctctgt
    10381 catcgttaca atcaacatgc taccctccgc gagatcatcc gtgtttcaaa cccggcagct
    10441 tagttgccgt tcttccgaat agcatcggta acatgagcaa agtctgccgc cttacaacgg
    10501 ctctcccgct gacgccgtcc cggactgatg ggctgcctgt atcgagtggt gattttgtgc
    10561 cgagctgccg gtcggggagc tgttggctgg ctggtggcag gatatattgt ggtgtaaaca
    10621 aattgacgct tagacaactt aataacacat tgcggacgtt tttaatgtac tgaattaacg
    10681 ccgaattaat tcgggggatc tggattttag tactggattt tggttttagg aattagaaat
    10741 tttattgata gaagtatttt acaaatacaa atacatacta agggtttctt atatgctcaa
    10801 cacatgagcg aaaccctata ggaaccctaa ttcccttatc tgggaactac tcacacatta
    10861 ttatggagaa actcgagctt gtcgatcgac agatcccggt cggcatctac tctatttctt
    10921 tgccctcgga cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc
    10981 atcggtccag acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc
    11041 ggatcggacg attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc
    11101 aaccaagctc tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagtcgtgg
    11161 cgatcctgca agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc
    11221 caaccacggc ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc
    11281 ctcgctccag tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc
    11341 gaaatccgcg tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc
    11401 atcgagagcc tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata
    11461 cacatgggga tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc
    11521 ttgcggtccg aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc
    11581 catagcctcc gcgaccggtt gtagaacagc gggcagttcg gtttcaggca ggtcttgcaa
    11641 cgtgacaccc tgtgaacggc gggagatgca ataggtcagg ctctcgctaa actccccaat
    11701 gtcaagcact tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc
    11761 tttgtagaaa ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc
    11821 gaagctgaaa gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc
    11881 gaacttttcg atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttcatatc
    11941 tcattgcccc ccggatctgc gaaagctcga gagagataga tttgtagaga gagactggtg
    12001 atttcagcgt gtcctctcca aatgaaatga acttccttat atagaggaag ggtcttgcga
    12061 aggatagtgg gattgtgcgt catcccttac gtcagtggag atatcacatc aatccacttg
    12121 ctttgaagac gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc
    12181 atctttggga ccactgtcgg cagaggcatc ttgaacgata gcctttcctt tatcgcaatg
    12241 atggcatttg taggtgccac cttccttttc tactgtcctt ttgatgaagt gacagatagc
    12301 tgggcaatgg aatccgagga ggtttcccga tattaccctt tgttgaaaag tctcaatagc
    12361 cctttggtct tctgagactg tatctttgat attcttggag tagacgagag tgtcgtgctc
    12421 caccatgttc acatcaatcc acttgctttg aagacgtggt tggaacgtct tctttttcca
    12481 cgatgctcct cgtgggtggg ggtccatctt tgggaccact gtcggcagag gcatcttgaa
    12541 cgatagcctt tcctttatcg caatgatggc atttgtaggt gccaccttcc ttttctactg
    12601 tccttttgat gaagtgacag atagctgggc aatggaatcc gaggaggttt cccgatatta
    12661 ccctttgttg aaaagtctca atagcccttt ggtcttctga gactgtatct ttgatattct
    12721 tggagtagac gagagtgtcg tgctccacca tgttggcaag ctgctctagc caatacgcaa
    12781 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
    12841 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
    12901 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
    12961 atttcacaca ggaaacagct atgacatgat tacgaattcg agct
//
