LOCUS       sequence-379570-       13912 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 13912)
  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 13912)
  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-MCS35S
FEATURES             Location/Qualifiers
     source          1..13912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(9..261)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(334..639)
                     /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     complement(436..455)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     promoter        complement(670..772)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     promoter        complement(968..1313)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1828..1845)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1828..1844)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1837..1859)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        2221..2566
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             2598..3308
                     /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(2625..2653)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3248..3268
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             3315..5120
                     /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             5127..5144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      5176..5428
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    5450..5474
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6775..7404
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             7833..8906
                     /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      8972..9166
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(9402..9424)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    9510..9650
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     9565..9584
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(9665..9682)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9836..10424)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9916..9935)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(10511..11305)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    11730..11754
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    11832..12006
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(12063..12860)
                     /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(12174..12193)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     12784..12803
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(12923..13599)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    13790..13811
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13826..13831,13832..13849,13850..13856)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13864..13880
                     /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     13869..13891
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13888..13904
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13888..13904
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gaattcccga tctagtaaca tagatgacac cgcgcgcgat aatttatcct agtttgcgcg
       61 ctatattttg ttttctatcg cgtattaaat gtataattgc gggactctaa tcataaaaac
      121 ccatctcata aataacgtca tgcattacat gttaattatt acatgcttaa cgtaattcaa
      181 cagaaattat atgataatca tcgcaagacc ggcaacagga ttcaatctta agaaacttta
      241 ttgccaaatg tttgaacgat cggggaaatt cgagctcggt ctcagcagac cacaagtggc
      301 cagactggcc ctgtgtataa gggagcctga catttatatt ccccagaaca tcaggttaat
      361 ggcgtttttg atgtcatttt cgcggtggct gagatcagcc acttcttccc cgataacgga
      421 taccggcaca ctggccatat cggtggtcat catgcgccag ctttcatccc cgatatgcac
      481 caccgggtaa agttcacggg agactttatc tgacagcaga cgtgcactgg ccagggggat
      541 caccatccgt cgcccaggcg tgtcaataat atcactctgt acatccacaa acagacgata
      601 acggctctct cttttatagg tgtaaacctt aaactgcatt tcaccagtcc ctgttctcgt
      661 cagcaaaagt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
      721 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca
      781 ttttcgccaa aagttggccc agggcttccc ggtatcaaca gggacaccag gatttattta
      841 ttctgcgaag tgatcttccg tcacaggtat ttattcggcg caaagtgcgt cgggtgatgc
      901 tgccaactta gtcgagaatg gccgacgagg ccaggactgg acagagacca ctctctagag
      961 tcccccgtgt tctctccaaa tgaaatgaac ttccttatat agaggaaggg tcttgcgaag
     1021 gatagtggga ttgtgcgtca tcccttacgt cagtggagat atcacatcaa tccacttgct
     1081 ttgaagacgt ggttggaacg tcttcttttt ccacgatgct cctcgtgggt gggggtccat
     1141 ctttgggacc actgtcggca gaggcatctt caacgatggc ctttccttta tcgcaatgat
     1201 ggcatttgta ggagccacct tccttttcca ctatcttcac aataaagtga cagatagctg
     1261 ggcaatggaa tccgaggagg tttccggata ttaccctttg ttgaaaagtc tcaattgccc
     1321 tttggtcttc tgagactgta tctttgatat ttttggagta gacaagtgtg tcgtgctcca
     1381 ccatgttgac gaagattttc ttcttgtcat tgagtcgtaa gagactctgt atgaactgtt
     1441 cgccagtctt tacggcgagt tctgttaggt cctctatttg aatctttgac tccatggcct
     1501 ttgattcagt gggaactacc tttttagaga ctccaatctc tattacttgc cttggtttgt
     1561 gaagcaagcc ttgaatcgtc catactggaa tagtacttct gatcttgaga aatatatctt
     1621 tctctgtgtt cttgatgcag ttagtcctga atcttttgac tgcatcttta accttcttgg
     1681 gaaggtattt gatttcctgg agattattgc tcgggtagat cgtcttgatg agacctgctg
     1741 cgtaagcctc tctaaccatc tgtgggttag cattctttct gaaattgaaa aggctaatct
     1801 ggggacctgc aggcatgcaa gcttggcact ggccgtcgtt ttacaacgtc gtgactggga
     1861 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     1921 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     1981 atgctagagc agcttgagct tggatcagat tgtcgtttcc cgccttcagt ttagcttcat
     2041 ggagtcaaag attcaaatag aggacctaac agaactcgcc gtaaagactg gcgaacagtt
     2101 catacagagt ctcttacgac tcaatgacaa gaagaaaatc ttcgtcaaca tggtggagca
     2161 cgacacactt gtctactcca aaaatatcaa agatacagtc tcagaagacc aaagggcaat
     2221 tgagactttt caacaaaggg taatatccgg aaacctcctc ggattccatt gcccagctat
     2281 ctgtcacttt attgtgaaga tagtggaaaa ggaaggtggc tcctacaaat gccatcattg
     2341 cgataaagga aaggccatcg ttgaagatgc ctctgccgac agtggtccca aagatggacc
     2401 cccacccacg aggagcatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt
     2461 ggattgatgt gatatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca
     2521 agacccttcc tctatataag gaagttcatt tcatttggag agaacacggg ggactcttga
     2581 ccatggtaga tctgactagt aaaggagaag aacttttcac tggagttgtc ccaattcttg
     2641 ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag ggtgaaggtg
     2701 atgcaacata cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc
     2761 cgtggccaac acttgtcact actttctctt atggtgttca atgcttttca agatacccag
     2821 atcatatgaa gcggcacgac ttcttcaaga gcgccatgcc tgagggatac gtgcaggaga
     2881 ggaccatctt cttcaaggac gacgggaact acaagacacg tgctgaagtc aagtttgagg
     2941 gagacaccct cgtcaacagg atcgagctta agggaatcga tttcaaggag gacggaaaca
     3001 tcctcggcca caagttggaa tacaactaca actcccacaa cgtatacatc atggccgaca
     3061 agcaaaagaa cggcatcaaa gccaacttca agacccgcca caacatcgaa gacggcggcg
     3121 tgcaactcgc tgatcattat caacaaaata ctccaattgg cgatggccct gtccttttac
     3181 cagacaacca ttacctgtcc acacaatctg ccctttcgaa agatcccaac gaaaagagag
     3241 accacatggt ccttcttgag tttgtaacag ctgctgggat tacacatggc atggatgaac
     3301 tatacaaagc tagtttacgt cctgtagaaa ccccaacccg tgaaatcaaa aaactcgacg
     3361 gcctgtgggc attcagtctg gatcgcgaaa actgtggaat tgatcagcgt tggtgggaaa
     3421 gcgcgttaca agaaagccgg gcaattgctg tgccaggcag ttttaacgat cagttcgccg
     3481 atgcagatat tcgtaattat gcgggcaacg tctggtatca gcgcgaagtc tttataccga
     3541 aaggttgggc aggccagcgt atcgtgctgc gtttcgatgc ggtcactcat tacggcaaag
     3601 tgtgggtcaa taatcaggaa gtgatggagc atcagggcgg ctatacgcca tttgaagccg
     3661 atgtcacgcc gtatgttatt gccgggaaaa gtgtacgtat caccgtttgt gtgaacaacg
     3721 aactgaactg gcagactatc ccgccgggaa tggtgattac cgacgaaaac ggcaagaaaa
     3781 agcagtctta cttccatgat ttctttaact atgccggaat ccatcgcagc gtaatgctct
     3841 acaccacgcc gaacacctgg gtggacgata tcaccgtggt gacgcatgtc gcgcaagact
     3901 gtaaccacgc gtctgttgac tggcaggtgg tggccaatgg tgatgtcagc gttgaactgc
     3961 gtgatgcgga tcaacaggtg gttgcaactg gacaaggcac tagcgggact ttgcaagtgg
     4021 tgaatccgca cctctggcaa ccgggtgaag gttatctcta tgaactgtgc gtcacagcca
     4081 aaagccagac agagtgtgat atctacccgc ttcgcgtcgg catccggtca gtggcagtga
     4141 agggcgaaca gttcctgatt aaccacaaac cgttctactt tactggcttt ggtcgtcatg
     4201 aagatgcgga cttgcgtggc aaaggattcg ataacgtgct gatggtgcac gaccacgcat
     4261 taatggactg gattggggcc aactcctacc gtacctcgca ttacccttac gctgaagaga
     4321 tgctcgactg ggcagatgaa catggcatcg tggtgattga tgaaactgct gctgtcggct
     4381 ttcagctgtc tttaggcatt ggtttcgaag cgggcaacaa gccgaaagaa ctgtacagcg
     4441 aagaggcagt caacggggaa actcagcaag cgcacttaca ggcgattaaa gagctgatag
     4501 cgcgtgacaa aaaccaccca agcgtggtga tgtggagtat tgccaacgaa ccggataccc
     4561 gtccgcaagg tgcacgggaa tatttcgcgc cactggcgga agcaacgcgt aaactcgacc
     4621 cgacgcgtcc gatcacctgc gtcaatgtaa tgttctgcga cgctcacacc gataccatca
     4681 gcgatctctt tgatgtgctg tgcctgaacc gttattacgg atggtatgtc caaagcggcg
     4741 atttggaaac ggcagagaag gtactggaaa aagaacttct ggcctggcag gagaaactgc
     4801 atcagccgat tatcatcacc gaatacggcg tggatacgtt agccgggctg cactcaatgt
     4861 acaccgacat gtggagtgaa gagtatcagt gtgcatggct ggatatgtat caccgcgtct
     4921 ttgatcgcgt cagcgccgtc gtcggtgaac aggtatggaa tttcgccgat tttgcgacct
     4981 cgcaaggcat attgcgcgtt ggcggtaaca agaaagggat cttcactcgc gaccgcaaac
     5041 cgaagtcggc ggcttttctg ctgcaaaaac gctggactgg catgaacttc ggtgaaaaac
     5101 cgcagcaggg aggcaaacaa gctagccacc accaccacca ccacgtgtga attggtgacc
     5161 agctcgaatt tccccgatcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct
     5221 gttgccggtc ttgcgatgat tatcatataa tttctgttga attacgttaa gcatgtaata
     5281 attaacatgt aatgcatgac gttatttatg agatgggttt ttatgattag agtcccgcaa
     5341 ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg
     5401 cgcgcggtgt catctatgtt actagatcgg gaattaaact atcagtgttt gacaggatat
     5461 attggcgggt aaacctaaga gaaaagagcg tttattagaa taatcggata tttaaaaggg
     5521 cgtgaaaagg tttatccgtt cgtccatttg tatgtgcatg ccaaccacag ggttcccctc
     5581 gggatcaaag tactttgatc caacccctcc gctgctatag tgcagtcggc ttctgacgtt
     5641 cagtgcagcc gtcttctgaa aacgacatgt cgcacaagtc ctaagttacg cgacaggctg
     5701 ccgccctgcc cttttcctgg cgttttcttg tcgcgtgttt tagtcgcata aagtagaata
     5761 cttgcgacta gaaccggaga cattacgcca tgaacaagag cgccgccgct ggcctgctgg
     5821 gctatgcccg cgtcagcacc gacgaccagg acttgaccaa ccaacgggcc gaactgcacg
     5881 cggccggctg caccaagctg ttttccgaga agatcaccgg caccaggcgc gaccgcccgg
     5941 agctggccag gatgcttgac cacctacgcc ctggcgacgt tgtgacagtg accaggctag
     6001 accgcctggc ccgcagcacc cgcgacctac tggacattgc cgagcgcatc caggaggccg
     6061 gcgcgggcct gcgtagcctg gcagagccgt gggccgacac caccacgccg gccggccgca
     6121 tggtgttgac cgtgttcgcc ggcattgccg agttcgagcg ttccctaatc atcgaccgca
     6181 cccggagcgg gcgcgaggcc gccaaggcgc gaggcgtgaa gtttggcccc cgccctaccc
     6241 tcaccccggc acagatcgcg cacgcccgcg agctgatcga ccaggaaggc cgcaccgtga
     6301 aagaggcggc tgcactgctt ggcgtgcatc gctcgaccct gtaccgcgca cttgagcgca
     6361 gcgaggaagt gacgcccacc gaggccaggc ggcgcggtgc cttccgtgag gacgcattga
     6421 ccgaggccga cgccctggcg gccgccgaga atgaacgcca agaggaacaa gcatgaaacc
     6481 gcaccaggac ggccaggacg aaccgttttt cattaccgaa gagatcgagg cggagatgat
     6541 cgcggccggg tacgtgttcg agccgcccgc gcacgtctca accgtgcggc tgcatgaaat
     6601 cctggccggt ttgtctgatg ccaagctggc ggcctggccg gcgagcttgg ccgctgaaga
     6661 aaccgagcgc cgccgtctaa aaaggtgatg tgtatttgag taaaacagct tgcgtcatgc
     6721 ggtcgctgcg tatatgatgc gatgagtaaa taaacaaata cgcaagggga acgcatgaag
     6781 gttatcgctg tacttaacca gaaaggcggg tcaggcaaga cgaccatcgc aacccatcta
     6841 gcccgcgccc tgcaactcgc cggggccgat gttctgttag tcgattccga tccccagggc
     6901 agtgcccgcg attgggcggc cgtgcgggaa gatcaaccgc taaccgttgt cggcatcgac
     6961 cgcccgacga ttgaccgcga cgtgaaggcc atcggccggc gcgacttcgt agtgatcgac
     7021 ggagcgcccc aggcggcgga cttggctgtg tccgcgatca aggcagccga cttcgtgctg
     7081 attccggtgc agccaagccc ttacgacata tgggccaccg ccgacctggt ggagctggtt
     7141 aagcagcgca ttgaggtcac ggatggaagg ctacaagcgg cctttgtcgt gtcgcgggcg
     7201 atcaaaggca cgcgcatcgg cggtgaggtt gccgaggcgc tggccgggta cgagctgccc
     7261 attcttgagt cccgtatcac gcagcgcgtg agctacccag gcactgccgc cgccggcaca
     7321 accgttcttg aatcagaacc cgagggcgac gctgcccgcg aggtccaggc gctggccgct
     7381 gaaattaaat caaaactcat ttgagttaat gaggtaaaga gaaaatgagc aaaagcacaa
     7441 acacgctaag tgccggccgt ccgagcgcac gcagcagcaa ggctgcaacg ttggccagcc
     7501 tggcagacac gccagccatg aagcgggtca actttcagtt gccggcggag gatcacacca
     7561 agctgaagat gtacgcggta cgccaaggca agaccattac cgagctgcta tctgaataca
     7621 tcgcgcagct accagagtaa atgagcaaat gaataaatga gtagatgaat tttagcggct
     7681 aaaggaggcg gcatggaaaa tcaagaacaa ccaggcaccg acgccgtgga atgccccatg
     7741 tgtggaggaa cgggcggttg gccaggcgta agcggctggg ttgcctgccg gccctgcaat
     7801 ggcactggaa cccccaagcc cgaggaatcg gcgtgagcgg tcgcaaacca tccggcccgg
     7861 tacaaatcgg cgcggcgctg ggtgatgacc tggtggagaa gttgaaggcg gcgcaggccg
     7921 cccagcggca acgcatcgag gcagaagcac gccccggtga atcgtggcaa gcggccgctg
     7981 atcgaatccg caaagaatcc cggcaaccgc cggcagccgg tgcgccgtcg attaggaagc
     8041 cgcccaaggg cgacgagcaa ccagattttt tcgttccgat gctctatgac gtgggcaccc
     8101 gcgatagtcg cagcatcatg gacgtggccg ttttccgtct gtcgaagcgt gaccgacgag
     8161 ctggcgaggt gatccgctac gagcttccag acgggcacgt agaggtttcc gcagggccgg
     8221 cgggcatggc cagtgtgtgg gattacgacc tggtactgat ggcggtttcc catctaaccg
     8281 aatccatgaa ccgataccgg gaagggaagg gagacaagcc cggccgcgtg ttccgtccac
     8341 acgttgcgga cgtactcaag ttctgccggc gagccgatgg cggaaagcag aaagacgacc
     8401 tggtagaaac ctgcattcgg ttaaacacca cgcacgttgc catgcagcgt acgaagaagg
     8461 ccaagaacgg ccgcctggtg acggtatccg agggtgaagc cttgattagc cgctacaaga
     8521 tcgtaaagag cgaaaccggg cggccggagt acatcgagat cgagctagct gattggatgt
     8581 accgcgagat cacagaaggc aagaacccgg acgtgctgac ggttcacccc gattactttt
     8641 tgatcgatcc cggcatcggc cgttttctct accgcctggc acgccgcgcc gcaggcaagg
     8701 cagaagccag atggttgttc aagacgatct acgaacgcag tggcagcgcc ggagagttca
     8761 agaagttctg tttcaccgtg cgcaagctga tcgggtcaaa tgacctgccg gagtacgatt
     8821 tgaaggagga ggcggggcag gctggcccga tcctagtcat gcgctaccgc aacctgatcg
     8881 agggcgaagc atccgccggt tcctaatgta cggagcagat gctagggcaa attgccctag
     8941 caggggaaaa aggtcgaaaa ggtctctttc ctgtggatag cacgtacatt gggaacccaa
     9001 agccgtacat tgggaaccgg aacccgtaca ttgggaaccc aaagccgtac attgggaacc
     9061 ggtcacacat gtaagtgact gatataaaag agaaaaaagg cgatttttcc gcctaaaact
     9121 ctttaaaact tattaaaact cttaaaaccc gcctggcctg tgcataactg tctggccagc
     9181 gcacagccga agagctgcaa aaagcgccta cccttcggtc gctgcgctcc ctacgccccg
     9241 ccgcttcgcg tcggcctatc gcggccgctg gccgctcaaa aatggctggc ctacggccag
     9301 gcaatctacc agggcgcgga caagccgcgc cgtcgccact cgaccgccgg cgcccacatc
     9361 aaggcaccct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     9421 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     9481 gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc
     9541 ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     9601 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg
     9661 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     9721 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     9781 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     9841 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     9901 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     9961 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
    10021 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
    10081 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
    10141 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
    10201 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
    10261 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
    10321 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
    10381 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
    10441 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatgc
    10501 attctaggta ctaaaacaat tcatccagta aaatataata ttttattttc tcccaatcag
    10561 gcttgatccc cagtaagtca aaaaatagct cgacatactg ttcttccccg atatcctccc
    10621 tgatcgaccg gacgcagaag gcaatgtcat accacttgtc cgccctgccg cttctcccaa
    10681 gatcaataaa gccacttact ttgccatctt tcacaaagat gttgctgtct cccaggtcgc
    10741 cgtgggaaaa gacaagttcc tcttcgggct tttccgtctt taaaaaatca tacagctcgc
    10801 gcggatcttt aaatggagtg tcttcttccc agttttcgca atccacatcg gccagatcgt
    10861 tattcagtaa gtaatccaat tcggctaagc ggctgtctaa gctattcgta tagggacaat
    10921 ccgatatgtc gatggagtga aagagcctga tgcactccgc atacagctcg ataatctttt
    10981 cagggctttg ttcatcttca tactcttccg agcaaaggac gccatcggcc tcactcatga
    11041 gcagattgct ccagccatca tgccgttcaa agtgcaggac ctttggaaca ggcagctttc
    11101 cttccagcca tagcatcatg tccttttccc gttccacatc ataggtggtc cctttatacc
    11161 ggctgtccgt catttttaaa tataggtttt cattttctcc caccagctta tataccttag
    11221 caggagacat tccttccgta tcttttacgc agcggtattt ttcgatcagt tttttcaatt
    11281 ccggtgatat tctcatttta gccatttatt atttccttcc tcttttctac agtatttaaa
    11341 gataccccaa gaagctaatt ataacaagac gaactccaat tcactgttcc ttgcattcta
    11401 aaaccttaaa taccagaaaa cagctttttc aaagttgttt tcaaagttgg cgtataacat
    11461 agtatcgacg gagccgattt tgaaaccgcg gtgatcacag gcagcaacgc tctgtcatcg
    11521 ttacaatcaa catgctaccc tccgcgagat catccgtgtt tcaaacccgg cagcttagtt
    11581 gccgttcttc cgaatagcat cggtaacatg agcaaagtct gccgccttac aacggctctc
    11641 ccgctgacgc cgtcccggac tgatgggctg cctgtatcga gtggtgattt tgtgccgagc
    11701 tgccggtcgg ggagctgttg gctggctggt ggcaggatat attgtggtgt aaacaaattg
    11761 acgcttagac aacttaataa cacattgcgg acgtttttaa tgtactgaat taacgccgaa
    11821 ttaattcggg ggatctggat tttagtactg gattttggtt ttaggaatta gaaattttat
    11881 tgatagaagt attttacaaa tacaaataca tactaagggt ttcttatatg ctcaacacat
    11941 gagcgaaacc ctataggaac cctaattccc ttatctggga actactcaca cattattatg
    12001 gagaaactcg agcttgtcga tcgactctag ctagaggatc gatccgaacc ccagagtccc
    12061 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
    12121 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
    12181 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg
    12241 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgtgtc
    12301 acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc
    12361 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
    12421 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
    12481 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
    12541 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
    12601 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
    12661 cgcgctgcct cgtcctggag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
    12721 accgggcgcc cctgcgctga cagccgaaac acggcggcat cagagcagcc gattgtctgt
    12781 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
    12841 ccatcttgtt caatccccat ggtcgatcga cagatctgcg aaagctcgag agagatagat
    12901 ttgtagagag agactggtga tttcagcgtg tcctctccaa atgaaatgaa cttccttata
    12961 tagaggaagg gtcttgcgaa ggatagtggg attgtgcgtc atcccttacg tcagtggaga
    13021 tatcacatca atccacttgc tttgaagacg tggttggaac gtcttctttt tccacgatgc
    13081 tcctcgtggg tgggggtcca tctttgggac cactgtcggc agaggcatct tgaacgatag
    13141 cctttccttt atcgcaatga tggcatttgt aggtgccacc ttccttttct actgtccttt
    13201 tgatgaagtg acagatagct gggcaatgga atccgaggag gtttcccgat attacccttt
    13261 gttgaaaagt ctcaatagcc ctttggtctt ctgagactgt atctttgata ttcttggagt
    13321 agacgagagt gtcgtgctcc accatgttca catcaatcca cttgctttga agacgtggtt
    13381 ggaacgtctt ctttttccac gatgctcctc gtgggtgggg gtccatcttt gggaccactg
    13441 tcggcagagg catcttgaac gatagccttt cctttatcgc aatgatggca tttgtaggtg
    13501 ccaccttcct tttctactgt ccttttgatg aagtgacaga tagctgggca atggaatccg
    13561 aggaggtttc ccgatattac cctttgttga aaagtctcaa tagccctttg gtcttctgag
    13621 actgtatctt tgatattctt ggagtagacg agagtgtcgt gctccaccat gttggcaagc
    13681 tgctctagcc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
    13741 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
    13801 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
    13861 gaattgtgag cggataacaa tttcacacag gaaacagcta tgacatgatt ac
//
