LOCUS       sequence-380035-       14252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nimble cloning-compatible vector for plant BiFC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14252)
  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 14252)
  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-BiFC-VD3
FEATURES             Location/Qualifiers
     source          1..14252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      complement(111..363)
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     promoter        complement(1238..1618)
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     promoter        2614..2959
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    2989..3043
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     promoter        3225..3327
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3358..3663
                     /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"
     CDS             join(3736..3738,3739..3741,3742..4254)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(3781..3802)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4042..4061)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     terminator      4401..5108
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        6109..6454
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    6483..6537
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     primer_bind     6565..6586
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6641..6664
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             6674..6697
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6704..6727
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        6924..7026
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             7057..7362
                     /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"
     terminator      7451..7703
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(7754..7771)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7754..7770)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7763..7785)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    7973..7997
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             9298..9927
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             10356..11429
                     /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      11495..11689
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(11925..11947)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    12033..12173
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     12088..12107
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(12188..12205)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12359..12947)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12439..12458)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(13034..13828)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg
       61 gacgttttta atgtactgaa ttaacgccga attaattcga gctcggatct gataatttat
      121 ttgaaaattc ataagaaaag caaacgttac atgaattgat gaaacaatac aaagacagat
      181 aaagccacgc acatttagga tattggccga gattactgaa tattgagtaa gatcacggaa
      241 tttctgacag gagcatgtct tcaattcagc ccaaatggca gttgaaatac tcaaaccgcc
      301 ccatatgcag gagcggatca ttcattgttt gtttggttgc ctttgccaac atgggagtcc
      361 aagattctgc agtcaaatct cggtgacggg caggaccgga cggggcggta cctcacttgt
      421 acagctcgtc catgccgaga gtaattccag cagctgtaac aaattccaaa agaaccatgt
      481 gatctctctt ttcattaggg tctttagaca acttagattg agtagacaaa tagtggttat
      541 caggcaaaag aactggtcca tcaccaatag gagtattttg ctgataatga tcagccaatt
      601 gaactccacc atcttcaata ttatgtctaa tcttaaaatt agctttaatt ccattctttt
      661 gtttatcagc agtaatataa acattatctg aaaagtaatt atactccaac ttatgaccca
      721 aaatatttcc atcttcttta aagtcaatac ccttcaactc aattctatta accaaagtat
      781 ctccttcaaa cttcacttca gctctagtct tataatttcc atcgtcctta aagaagatag
      841 ttctctcttg aacatatcct tcaggcatag cagacttaaa gaaatcatgt tgtttcatgt
      901 ggtcagggta tctagcaaaa cattgaactc cccaagacaa agttgtcaca agagtcggcc
      961 aaggaaccgg caactttcca gttgtacaaa taaacttcaa agtgagcttt ccataagtag
     1021 catctccctc accctcacca ctaacagaaa acttatgtcc attaacgtcg ccatccaatt
     1081 caacaagaat aggcacaact ccagtaaaca gctcctcgcc cttgctcact gagaaagatg
     1141 aactgtggaa cacgaagtag atgataagga acaaactgtt gagaagcaac atatacagga
     1201 atatcttcag aatcttaggg ttccgcttac tcattcgcga tttggtgtat cgagattggt
     1261 tatgaaattc agatgctagt gtaatgtatt ggtaatttgg gaagatataa taggaagcaa
     1321 ggctatttat ccatttctga aaaggcgaaa tggcgtcacc gcgagcgtca cgcgcattcc
     1381 gttcttgctg taaagcgttg tttggtacac ttttgactag cgaggcttgg cgtgtcagcg
     1441 tatctattca aaagtcgtta atggctgcgg atcaagaaaa agttggaata gaaacagaat
     1501 acccgcgaaa ttcaggcccg gttgccatgt cctacacgcc gaaataaacg accaaattag
     1561 tagaaaaata aaaactgact cggatactta cgtcacgtct tgcgcactga tttgaaaaat
     1621 ctcagaattc caatcccaca aaaatctgag cttaacagca cagttgctcc tctcagagca
     1681 gaatcgggta ttcaacaccc tcatatcaac tactacgttg tgtataacgg tccacatgcc
     1741 ggtatatacg atgactgggg ttgtacaaag gcggcaacaa acggcgttcc cggagttgca
     1801 cacaagaaat ttgccactat tacagaggca agagcagcag ctgacgcgta cacaacaagt
     1861 cagcaaacag acaggttgaa cttcatcccc aaaggagaag ctcaactcaa gcccaagagc
     1921 tttgctaagg ccctaacaag cccaccaaag caaaaagccc actggctcac gctaggaacc
     1981 aaaaggccca gcagtgatcc agccccaaaa gagatctcct ttgccccgga gattacaatg
     2041 gacgatttcc tctatcttta cgatctagga aggaagttcg aaggtgaagg tgacgacact
     2101 atgttcacca ctgataatga gaaggttagc ctcttcaatt tcagaaagaa tgctgaccca
     2161 cagatggtta gagaggccta cgcagcaggt ctcatcaaga cgatctaccc gagtaacaat
     2221 ctccaggaga tcaaatacct tcccaagaag gttaaagatg cagtcaaaag attcaggact
     2281 aattgcatca agaacacaga gaaagacata tttctcaaga tcagaagtac tattccagta
     2341 tggacgattc aaggcttgct tcataaacca aggcaagtaa tagagattgg agtctctaaa
     2401 aaggtagttc ctactgaatc taaggccatg catggagtct aagattcaaa tcgaggatct
     2461 aacagaactc gccgtgaaga ctggcgaaca gttcatacag agtcttttac gactcaatga
     2521 caagaagaaa atcttcgtca acatggtgga gcacgacact ctggtctact ccaaaaatgt
     2581 caaagataca gtctcagaag accaaagggc tattgagact tttcaacaaa ggataatttc
     2641 gggaaacctc ctcggattcc attgcccagc tatctgtcac ttcatcgaaa ggacagtaga
     2701 aaaggaaggt ggctcctaca aatgccatca ttgcgataaa ggaaaggcta tcattcaaga
     2761 tctctctgcc gacagtggtc ccaaagatgg acccccaccc acgaggagca tcgtggaaaa
     2821 agaagacgtt ccaaccacgt cttcaaagca agtggattga tgtgacatct ccactgacgt
     2881 aagggatgac gcacaatccc actatccttc gcaagaccct tcctctatat aaggaagttc
     2941 atttcatttg gagaggacac gctcgagtat aagagctcta tttttacaac aattaccaac
     3001 aacaacaaac aacaaacaac attacaatta catttacaat taccagtggt ctctgtccag
     3061 tcctggcctc gtcggccatt ctcgactaag ttggcagcat cacccgacgc actttgcgcc
     3121 gaataaatac ctgtgacgga agatcacttc gcagaataaa taaatcctgg tgtccctgtt
     3181 gataccggga agccctgggc caacttttgg cgaaaatgag acgttgatcg gcacgtaaga
     3241 ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg
     3301 agattttcag gagctaagga agctaaactt ttgctgacga gaacagggac tggtgaaatg
     3361 cagtttaagg tttacaccta taaaagagag agccgttatc gtctgtttgt ggatgtacag
     3421 agtgatatta ttgacacgcc tgggcgacgg atggtgatcc ccctggccag tgcacgtctg
     3481 ctgtcagata aagtctcccg tgaactttac ccggtggtgc atatcgggga tgaaagctgg
     3541 cgcatgatga ccaccgatat ggccagtgtg ccggtatccg ttatcgggga agaagtggct
     3601 gatctcagcc accgcgaaaa tgacatcaaa aacgccatta acctgatgtt ctggggaata
     3661 taaatgtcag gctcccttat acacagggcc agtctggcca cttgtggtct gctgagaccc
     3721 ggaggtgggt caggaatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3781 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     3841 ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc
     3901 gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt cgcccgctac
     3961 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     4021 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     4081 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     4141 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc
     4201 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     4261 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     4321 ctgcccgaca accactacct gagctaccag tccaagctga gcaaataatc tagacccggg
     4381 ggttaattaa ccctagagtc ctgctttaat gagatatgcg agacgcctat gatcgcatga
     4441 tatttgcttt caattctgtt gtgcacgttg taaaaaacct gagcatgtgt agctcagatc
     4501 cttaccgccg gtttcggttc attctaatga atatatcacc cgttactatc gtatttttat
     4561 gaataatatt ctccgttcaa tttactgatt gtaccctact acttatatgt acaatattaa
     4621 aatgaaaaca atatattgtg ctgaataggt ttatagcgac atctatgata gagcgccaca
     4681 ataacaaaca attgcgtttt attattacaa atccaatttt aaaaaaagcg gcagaaccgg
     4741 tcaaacctaa aagactgatt acataaatct tattcaaatt tcaaaagtgc cccaggggct
     4801 agtatctacg acacaccgag cggcgaacta ataacgctca ctgaagggaa ctccggttcc
     4861 ccgccggcgc gcatgggtga gattccttga agttgagtat tggccgtccg ctctaccgaa
     4921 agttacgggc accattcaac ccggtccagc acggcggccg ggtaaccgac ttgctgcccc
     4981 gagaattatg cagcattttt ttggtgtatg tgggccccaa atgaagtgca ggtcaaacct
     5041 tgacagtgac gacaaatcgt tgggcgggtc cagggcgaat tttgcgacaa catgtcgagg
     5101 ctcagcagga cctgctcata tgttccaatc ccacaaaaat ctgagcttaa cagcacagtt
     5161 gctcctctca gagcagaatc gggtattcaa caccctcata tcaactacta cgttgtgtat
     5221 aacggtccac atgccggtat atacgatgac tggggttgta caaaggcggc aacaaacggc
     5281 gttcccggag ttgcacacaa gaaatttgcc actattacag aggcaagagc agcagctgac
     5341 gcgtacacaa caagtcagca aacagacagg ttgaacttca tccccaaagg agaagctcaa
     5401 ctcaagccca agagctttgc taaggcccta acaagcccac caaagcaaaa agcccactgg
     5461 ctcacgctag gaaccaaaag gcccagcagt gatccagccc caaaagagat ctcctttgcc
     5521 ccggagatta caatggacga tttcctctat ctttacgatc taggaaggaa gttcgaaggt
     5581 gaaggtgacg acactatgtt caccactgat aatgagaagg ttagcctctt caatttcaga
     5641 aagaatgctg acccacagat ggttagagag gcctacgcag caggtctcat caagacgatc
     5701 tacccgagta acaatctcca ggagatcaaa taccttccca agaaggttaa agatgcagtc
     5761 aaaagattca ggactaattg catcaagaac acagagaaag acatatttct caagatcaga
     5821 agtactattc cagtatggac gattcaaggc ttgcttcata aaccaaggca agtaatagag
     5881 attggagtct ctaaaaaggt agttcctact gaatctaagg ccatgcatgg agtctaagat
     5941 tcaaatcgag gatctaacag aactcgccgt gaagactggc gaacagttca tacagagtct
     6001 tttacgactc aatgacaaga agaaaatctt cgtcaacatg gtggagcacg acactctggt
     6061 ctactccaaa aatgtcaaag atacagtctc agaagaccaa agggctattg agacttttca
     6121 acaaaggata atttcgggaa acctcctcgg attccattgc ccagctatct gtcacttcat
     6181 cgaaaggaca gtagaaaagg aaggtggctc ctacaaatgc catcattgcg ataaaggaaa
     6241 ggctatcatt caagatctct ctgccgacag tggtcccaaa gatggacccc cacccacgag
     6301 gagcatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg attgatgtga
     6361 catctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag acccttcctc
     6421 tatataagga agttcatttc atttggagag gacacgcctc aggtacagag catattttta
     6481 caacaattac caacaacaac aaacaacaaa caacattaca attacattta caattacaat
     6541 ggaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     6601 cactctcggc atggacgagc tgtacaaggg atctgggtca tggtctcatc cacaattcga
     6661 gaagtcagga ggagattata aggatgatga tgataaggct tcttggtcac atcctcagtt
     6721 tgaaaagtct ggagcttcag gaagagttgg ctcgaccatg cgtggcctcg tcggccattc
     6781 tcgactaagt tggcagcatc acccgacgca ctttgcgccg aataaatacc tgtgacggaa
     6841 gatcacttcg cagaataaat aaatcctggt gtccctgttg ataccgggaa gccctgggcc
     6901 aacttttggc gaaaatgaga cgttgatcgg cacgtaagag gttccaactt tcaccataat
     6961 gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa
     7021 gctaaacttt tgctgacgag aacagggact ggtgaaatgc agtttaaggt ttacacctat
     7081 aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgcct
     7141 gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt
     7201 gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg
     7261 gccagtgtgc cggtatccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat
     7321 gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata
     7381 cacagggcca gtctggccag acctggatgt ttccagtgga cgtccggata agcgatcgcc
     7441 atttaaatag gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc
     7501 cggtcttgcg atgattatca tataatttct gttgaattac gttaagcatg taataattaa
     7561 catgtaatgc atgacgttat ttatgagatg ggtttttatg attagagtcc cgcaattata
     7621 catttaatac gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc
     7681 ggtgtcatct atgttactag atcgttaacg atatgtcgag gctcagcagg acctgcaggc
     7741 atgcaagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     7801 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     7861 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgc tagagcagct
     7921 tgagcttgga tcagattgtc gtttcccgcc ttcagtttaa actatcagtg tttgacagga
     7981 tatattggcg ggtaaaccta agagaaaaga gcgtttatta gaataatcgg atatttaaaa
     8041 gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc atgccaacca cagggttccc
     8101 ctcgggatca aagtactttg atccaacccc tccgctgcta tagtgcagtc ggcttctgac
     8161 gttcagtgca gccgtcttct gaaaacgaca tgtcgcacaa gtcctaagtt acgcgacagg
     8221 ctgccgccct gcccttttcc tggcgttttc ttgtcgcgtg ttttagtcgc ataaagtaga
     8281 atacttgcga ctagaaccgg agacattacg ccatgaacaa gagcgccgcc gctggcctgc
     8341 tgggctatgc ccgcgtcagc accgacgacc aggacttgac caaccaacgg gccgaactgc
     8401 acgcggccgg ctgcaccaag ctgttttccg agaagatcac cggcaccagg cgcgaccgcc
     8461 cggagctggc caggatgctt gaccacctac gccctggcga cgttgtgaca gtgaccaggc
     8521 tagaccgcct ggcccgcagc acccgcgacc tactggacat tgccgagcgc atccaggagg
     8581 ccggcgcggg cctgcgtagc ctggcagagc cgtgggccga caccaccacg ccggccggcc
     8641 gcatggtgtt gaccgtgttc gccggcattg ccgagttcga gcgttcccta atcatcgacc
     8701 gcacccggag cgggcgcgag gccgccaagg cgcgaggcgt gaagtttggc ccccgcccta
     8761 ccctcacccc ggcacagatc gcgcacgccc gcgagctgat cgaccaggaa ggccgcaccg
     8821 tgaaagaggc ggctgcactg cttggcgtgc atcgctcgac cctgtaccgc gcacttgagc
     8881 gcagcgagga agtgacgccc accgaggcca ggcggcgcgg tgccttccgt gaggacgcat
     8941 tgaccgaggc cgacgccctg gcggccgccg agaatgaacg ccaagaggaa caagcatgaa
     9001 accgcaccag gacggccagg acgaaccgtt tttcattacc gaagagatcg aggcggagat
     9061 gatcgcggcc gggtacgtgt tcgagccgcc cgcgcacgtc tcaaccgtgc ggctgcatga
     9121 aatcctggcc ggtttgtctg atgccaagct ggcggcctgg ccggcgagct tggccgctga
     9181 agaaaccgag cgccgccgtc taaaaaggtg atgtgtattt gagtaaaaca gcttgcgtca
     9241 tgcggtcgct gcgtatatga tgcgatgagt aaataaacaa atacgcaagg ggaacgcatg
     9301 aaggttatcg ctgtacttaa ccagaaaggc gggtcaggca agacgaccat cgcaacccat
     9361 ctagcccgcg ccctgcaact cgccggggcc gatgttctgt tagtcgattc cgatccccag
     9421 ggcagtgccc gcgattgggc ggccgtgcgg gaagatcaac cgctaaccgt tgtcggcatc
     9481 gaccgcccga cgattgaccg cgacgtgaag gccatcggcc ggcgcgactt cgtagtgatc
     9541 gacggagcgc cccaggcggc ggacttggct gtgtccgcga tcaaggcagc cgacttcgtg
     9601 ctgattccgg tgcagccaag cccttacgac atatgggcca ccgccgacct ggtggagctg
     9661 gttaagcagc gcattgaggt cacggatgga aggctacaag cggcctttgt cgtgtcgcgg
     9721 gcgatcaaag gcacgcgcat cggcggtgag gttgccgagg cgctggccgg gtacgagctg
     9781 cccattcttg agtcccgtat cacgcagcgc gtgagctacc caggcactgc cgccgccggc
     9841 acaaccgttc ttgaatcaga acccgagggc gacgctgccc gcgaggtcca ggcgctggcc
     9901 gctgaaatta aatcaaaact catttgagtt aatgaggtaa agagaaaatg agcaaaagca
     9961 caaacacgct aagtgccggc cgtccgagcg cacgcagcag caaggctgca acgttggcca
    10021 gcctggcaga cacgccagcc atgaagcggg tcaactttca gttgccggcg gaggatcaca
    10081 ccaagctgaa gatgtacgcg gtacgccaag gcaagaccat taccgagctg ctatctgaat
    10141 acatcgcgca gctaccagag taaatgagca aatgaataaa tgagtagatg aattttagcg
    10201 gctaaaggag gcggcatgga aaatcaagaa caaccaggca ccgacgccgt ggaatgcccc
    10261 atgtgtggag gaacgggcgg ttggccaggc gtaagcggct gggttgcctg ccggccctgc
    10321 aatggcactg gaacccccaa gcccgaggaa tcggcgtgag cggtcgcaaa ccatccggcc
    10381 cggtacaaat cggcgcggcg ctgggtgatg acctggtgga gaagttgaag gcggcgcagg
    10441 ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg tgaatcgtgg caagcggccg
    10501 ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc cggtgcgccg tcgattagga
    10561 agccgcccaa gggcgacgag caaccagatt ttttcgttcc gatgctctat gacgtgggca
    10621 cccgcgatag tcgcagcatc atggacgtgg ccgttttccg tctgtcgaag cgtgaccgac
    10681 gagctggcga ggtgatccgc tacgagcttc cagacgggca cgtagaggtt tccgcagggc
    10741 cggcgggcat ggccagtgtg tgggattacg acctggtact gatggcggtt tcccatctaa
    10801 ccgaatccat gaaccgatac cgggaaggga agggagacaa gcccggccgc gtgttccgtc
    10861 cacacgttgc ggacgtactc aagttctgcc ggcgagccga tggcggaaag cagaaagacg
    10921 acctggtaga aacctgcatt cggttaaaca ccacgcacgt tgccatgcag cgtacgaaga
    10981 aggccaagaa cggccgcctg gtgacggtat ccgagggtga agccttgatt agccgctaca
    11041 agatcgtaaa gagcgaaacc gggcggccgg agtacatcga gatcgagcta gctgattgga
    11101 tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct gacggttcac cccgattact
    11161 ttttgatcga tcccggcatc ggccgttttc tctaccgcct ggcacgccgc gccgcaggca
    11221 aggcagaagc cagatggttg ttcaagacga tctacgaacg cagtggcagc gccggagagt
    11281 tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc aaatgacctg ccggagtacg
    11341 atttgaagga ggaggcgggg caggctggcc cgatcctagt catgcgctac cgcaacctga
    11401 tcgagggcga agcatccgcc ggttcctaat gtacggagca gatgctaggg caaattgccc
    11461 tagcagggga aaaaggtcga aaaggtctct ttcctgtgga tagcacgtac attgggaacc
    11521 caaagccgta cattgggaac cggaacccgt acattgggaa cccaaagccg tacattggga
    11581 accggtcaca catgtaagtg actgatataa aagagaaaaa aggcgatttt tccgcctaaa
    11641 actctttaaa acttattaaa actcttaaaa cccgcctggc ctgtgcataa ctgtctggcc
    11701 agcgcacagc cgaagagctg caaaaagcgc ctacccttcg gtcgctgcgc tccctacgcc
    11761 ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc aaaaatggct ggcctacggc
    11821 caggcaatct accagggcgc ggacaagccg cgccgtcgcc actcgaccgc cggcgcccac
    11881 atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
    11941 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
    12001 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
    12061 agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
    12121 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
    12181 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
    12241 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
    12301 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
    12361 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
    12421 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
    12481 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
    12541 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
    12601 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
    12661 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
    12721 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
    12781 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
    12841 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
    12901 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
    12961 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
    13021 tgcattctag gtactaaaac aattcatcca gtaaaatata atattttatt ttctcccaat
    13081 caggcttgat ccccagtaag tcaaaaaata gctcgacata ctgttcttcc ccgatatcct
    13141 ccctgatcga ccggacgcag aaggcaatgt cataccactt gtccgccctg ccgcttctcc
    13201 caagatcaat aaagccactt actttgccat ctttcacaaa gatgttgctg tctcccaggt
    13261 cgccgtggga aaagacaagt tcctcttcgg gcttttccgt ctttaaaaaa tcatacagct
    13321 cgcgcggatc tttaaatgga gtgtcttctt cccagttttc gcaatccaca tcggccagat
    13381 cgttattcag taagtaatcc aattcggcta agcggctgtc taagctattc gtatagggac
    13441 aatccgatat gtcgatggag tgaaagagcc tgatgcactc cgcatacagc tcgataatct
    13501 tttcagggct ttgttcatct tcatactctt ccgagcaaag gacgccatcg gcctcactca
    13561 tgagcagatt gctccagcca tcatgccgtt caaagtgcag gacctttgga acaggcagct
    13621 ttccttccag ccatagcatc atgtcctttt cccgttccac atcataggtg gtccctttat
    13681 accggctgtc cgtcattttt aaatataggt tttcattttc tcccaccagc ttatatacct
    13741 tagcaggaga cattccttcc gtatctttta cgcagcggta tttttcgatc agttttttca
    13801 attccggtga tattctcatt ttagccattt attatttcct tcctcttttc tacagtattt
    13861 aaagataccc caagaagcta attataacaa gacgaactcc aattcactgt tccttgcatt
    13921 ctaaaacctt aaataccaga aaacagcttt ttcaaagttg ttttcaaagt tggcgtataa
    13981 catagtatcg acggagccga ttttgaaacc gcggtgatca caggcagcaa cgctctgtca
    14041 tcgttacaat caacatgcta ccctccgcga gatcatccgt gtttcaaacc cggcagctta
    14101 gttgccgttc ttccgaatag catcggtaac atgagcaaag tctgccgcct tacaacggct
    14161 ctcccgctga cgccgtcccg gactgatggg ctgcctgtat cgagtggtga ttttgtgccg
    14221 agctgccggt cggggagctg ttggctggct gg
//
