LOCUS       sequence-380040-       14245 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 14245)
  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 14245)
  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-VD4
FEATURES             Location/Qualifiers
     source          1..14245
                     /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)"
     promoter        6719..6821
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             6852..7157
                     /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             7236..7259
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             7269..7292
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7299..7322
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     7358..7379
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      7444..7696
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(7747..7764)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7747..7763)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7756..7778)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    7966..7990
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             9291..9920
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             10349..11422
                     /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      11488..11682
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(11918..11940)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    12026..12166
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     12081..12100
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(12181..12198)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12352..12940)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12432..12451)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(13027..13821)
                     /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 caattacaga
     6541 gttggctcga ccatgcgtgg cctcgtcggc cattctcgac taagttggca gcatcacccg
     6601 acgcactttg cgccgaataa atacctgtga cggaagatca cttcgcagaa taaataaatc
     6661 ctggtgtccc tgttgatacc gggaagccct gggccaactt ttggcgaaaa tgagacgttg
     6721 atcggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat
     6781 tttttgagtt atcgagattt tcaggagcta aggaagctaa acttttgctg acgagaacag
     6841 ggactggtga aatgcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt
     6901 ttgtggatgt acagagtgat attattgaca cgcctgggcg acggatggtg atccccctgg
     6961 ccagtgcacg tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg
     7021 gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggta tccgttatcg
     7081 gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga
     7141 tgttctgggg aatataaatg tcaggctccc ttatacacag ggccagtctg gccagacctg
     7201 gatgtttcca gtggtccgga ggttctggtg gagcttggtc tcatccacaa ttcgagaagt
     7261 caggaggaga ttataaggat gatgatgata aggcttcttg gtcacatcct cagtttgaaa
     7321 agggatctgg gtcagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     7381 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagcgatc gccatttaaa
     7441 taggatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt tgccggtctt
     7501 gcgatgatta tcatataatt tctgttgaat tacgttaagc atgtaataat taacatgtaa
     7561 tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt atacatttaa
     7621 tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg cgcggtgtca
     7681 tctatgttac tagatcgtta acgatatgtc gaggctcagc aggacctgca ggcatgcaag
     7741 cttggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     7801 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     7861 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgctagagca gcttgagctt
     7921 ggatcagatt gtcgtttccc gccttcagtt taaactatca gtgtttgaca ggatatattg
     7981 gcgggtaaac ctaagagaaa agagcgttta ttagaataat cggatattta aaagggcgtg
     8041 aaaaggttta tccgttcgtc catttgtatg tgcatgccaa ccacagggtt cccctcggga
     8101 tcaaagtact ttgatccaac ccctccgctg ctatagtgca gtcggcttct gacgttcagt
     8161 gcagccgtct tctgaaaacg acatgtcgca caagtcctaa gttacgcgac aggctgccgc
     8221 cctgcccttt tcctggcgtt ttcttgtcgc gtgttttagt cgcataaagt agaatacttg
     8281 cgactagaac cggagacatt acgccatgaa caagagcgcc gccgctggcc tgctgggcta
     8341 tgcccgcgtc agcaccgacg accaggactt gaccaaccaa cgggccgaac tgcacgcggc
     8401 cggctgcacc aagctgtttt ccgagaagat caccggcacc aggcgcgacc gcccggagct
     8461 ggccaggatg cttgaccacc tacgccctgg cgacgttgtg acagtgacca ggctagaccg
     8521 cctggcccgc agcacccgcg acctactgga cattgccgag cgcatccagg aggccggcgc
     8581 gggcctgcgt agcctggcag agccgtgggc cgacaccacc acgccggccg gccgcatggt
     8641 gttgaccgtg ttcgccggca ttgccgagtt cgagcgttcc ctaatcatcg accgcacccg
     8701 gagcgggcgc gaggccgcca aggcgcgagg cgtgaagttt ggcccccgcc ctaccctcac
     8761 cccggcacag atcgcgcacg cccgcgagct gatcgaccag gaaggccgca ccgtgaaaga
     8821 ggcggctgca ctgcttggcg tgcatcgctc gaccctgtac cgcgcacttg agcgcagcga
     8881 ggaagtgacg cccaccgagg ccaggcggcg cggtgccttc cgtgaggacg cattgaccga
     8941 ggccgacgcc ctggcggccg ccgagaatga acgccaagag gaacaagcat gaaaccgcac
     9001 caggacggcc aggacgaacc gtttttcatt accgaagaga tcgaggcgga gatgatcgcg
     9061 gccgggtacg tgttcgagcc gcccgcgcac gtctcaaccg tgcggctgca tgaaatcctg
     9121 gccggtttgt ctgatgccaa gctggcggcc tggccggcga gcttggccgc tgaagaaacc
     9181 gagcgccgcc gtctaaaaag gtgatgtgta tttgagtaaa acagcttgcg tcatgcggtc
     9241 gctgcgtata tgatgcgatg agtaaataaa caaatacgca aggggaacgc atgaaggtta
     9301 tcgctgtact taaccagaaa ggcgggtcag gcaagacgac catcgcaacc catctagccc
     9361 gcgccctgca actcgccggg gccgatgttc tgttagtcga ttccgatccc cagggcagtg
     9421 cccgcgattg ggcggccgtg cgggaagatc aaccgctaac cgttgtcggc atcgaccgcc
     9481 cgacgattga ccgcgacgtg aaggccatcg gccggcgcga cttcgtagtg atcgacggag
     9541 cgccccaggc ggcggacttg gctgtgtccg cgatcaaggc agccgacttc gtgctgattc
     9601 cggtgcagcc aagcccttac gacatatggg ccaccgccga cctggtggag ctggttaagc
     9661 agcgcattga ggtcacggat ggaaggctac aagcggcctt tgtcgtgtcg cgggcgatca
     9721 aaggcacgcg catcggcggt gaggttgccg aggcgctggc cgggtacgag ctgcccattc
     9781 ttgagtcccg tatcacgcag cgcgtgagct acccaggcac tgccgccgcc ggcacaaccg
     9841 ttcttgaatc agaacccgag ggcgacgctg cccgcgaggt ccaggcgctg gccgctgaaa
     9901 ttaaatcaaa actcatttga gttaatgagg taaagagaaa atgagcaaaa gcacaaacac
     9961 gctaagtgcc ggccgtccga gcgcacgcag cagcaaggct gcaacgttgg ccagcctggc
    10021 agacacgcca gccatgaagc gggtcaactt tcagttgccg gcggaggatc acaccaagct
    10081 gaagatgtac gcggtacgcc aaggcaagac cattaccgag ctgctatctg aatacatcgc
    10141 gcagctacca gagtaaatga gcaaatgaat aaatgagtag atgaatttta gcggctaaag
    10201 gaggcggcat ggaaaatcaa gaacaaccag gcaccgacgc cgtggaatgc cccatgtgtg
    10261 gaggaacggg cggttggcca ggcgtaagcg gctgggttgc ctgccggccc tgcaatggca
    10321 ctggaacccc caagcccgag gaatcggcgt gagcggtcgc aaaccatccg gcccggtaca
    10381 aatcggcgcg gcgctgggtg atgacctggt ggagaagttg aaggcggcgc aggccgccca
    10441 gcggcaacgc atcgaggcag aagcacgccc cggtgaatcg tggcaagcgg ccgctgatcg
    10501 aatccgcaaa gaatcccggc aaccgccggc agccggtgcg ccgtcgatta ggaagccgcc
    10561 caagggcgac gagcaaccag attttttcgt tccgatgctc tatgacgtgg gcacccgcga
    10621 tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg aagcgtgacc gacgagctgg
    10681 cgaggtgatc cgctacgagc ttccagacgg gcacgtagag gtttccgcag ggccggcggg
    10741 catggccagt gtgtgggatt acgacctggt actgatggcg gtttcccatc taaccgaatc
    10801 catgaaccga taccgggaag ggaagggaga caagcccggc cgcgtgttcc gtccacacgt
    10861 tgcggacgta ctcaagttct gccggcgagc cgatggcgga aagcagaaag acgacctggt
    10921 agaaacctgc attcggttaa acaccacgca cgttgccatg cagcgtacga agaaggccaa
    10981 gaacggccgc ctggtgacgg tatccgaggg tgaagccttg attagccgct acaagatcgt
    11041 aaagagcgaa accgggcggc cggagtacat cgagatcgag ctagctgatt ggatgtaccg
    11101 cgagatcaca gaaggcaaga acccggacgt gctgacggtt caccccgatt actttttgat
    11161 cgatcccggc atcggccgtt ttctctaccg cctggcacgc cgcgccgcag gcaaggcaga
    11221 agccagatgg ttgttcaaga cgatctacga acgcagtggc agcgccggag agttcaagaa
    11281 gttctgtttc accgtgcgca agctgatcgg gtcaaatgac ctgccggagt acgatttgaa
    11341 ggaggaggcg gggcaggctg gcccgatcct agtcatgcgc taccgcaacc tgatcgaggg
    11401 cgaagcatcc gccggttcct aatgtacgga gcagatgcta gggcaaattg ccctagcagg
    11461 ggaaaaaggt cgaaaaggtc tctttcctgt ggatagcacg tacattggga acccaaagcc
    11521 gtacattggg aaccggaacc cgtacattgg gaacccaaag ccgtacattg ggaaccggtc
    11581 acacatgtaa gtgactgata taaaagagaa aaaaggcgat ttttccgcct aaaactcttt
    11641 aaaacttatt aaaactctta aaacccgcct ggcctgtgca taactgtctg gccagcgcac
    11701 agccgaagag ctgcaaaaag cgcctaccct tcggtcgctg cgctccctac gccccgccgc
    11761 ttcgcgtcgg cctatcgcgg ccgctggccg ctcaaaaatg gctggcctac ggccaggcaa
    11821 tctaccaggg cgcggacaag ccgcgccgtc gccactcgac cgccggcgcc cacatcaagg
    11881 caccctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
    11941 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
    12001 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
    12061 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
    12121 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc
    12181 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
    12241 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
    12301 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
    12361 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
    12421 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
    12481 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
    12541 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
    12601 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
    12661 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
    12721 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
    12781 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
    12841 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
    12901 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
    12961 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgcattc
    13021 taggtactaa aacaattcat ccagtaaaat ataatatttt attttctccc aatcaggctt
    13081 gatccccagt aagtcaaaaa atagctcgac atactgttct tccccgatat cctccctgat
    13141 cgaccggacg cagaaggcaa tgtcatacca cttgtccgcc ctgccgcttc tcccaagatc
    13201 aataaagcca cttactttgc catctttcac aaagatgttg ctgtctccca ggtcgccgtg
    13261 ggaaaagaca agttcctctt cgggcttttc cgtctttaaa aaatcataca gctcgcgcgg
    13321 atctttaaat ggagtgtctt cttcccagtt ttcgcaatcc acatcggcca gatcgttatt
    13381 cagtaagtaa tccaattcgg ctaagcggct gtctaagcta ttcgtatagg gacaatccga
    13441 tatgtcgatg gagtgaaaga gcctgatgca ctccgcatac agctcgataa tcttttcagg
    13501 gctttgttca tcttcatact cttccgagca aaggacgcca tcggcctcac tcatgagcag
    13561 attgctccag ccatcatgcc gttcaaagtg caggaccttt ggaacaggca gctttccttc
    13621 cagccatagc atcatgtcct tttcccgttc cacatcatag gtggtccctt tataccggct
    13681 gtccgtcatt tttaaatata ggttttcatt ttctcccacc agcttatata ccttagcagg
    13741 agacattcct tccgtatctt ttacgcagcg gtatttttcg atcagttttt tcaattccgg
    13801 tgatattctc attttagcca tttattattt ccttcctctt ttctacagta tttaaagata
    13861 ccccaagaag ctaattataa caagacgaac tccaattcac tgttccttgc attctaaaac
    13921 cttaaatacc agaaaacagc tttttcaaag ttgttttcaa agttggcgta taacatagta
    13981 tcgacggagc cgattttgaa accgcggtga tcacaggcag caacgctctg tcatcgttac
    14041 aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc ttagttgccg
    14101 ttcttccgaa tagcatcggt aacatgagca aagtctgccg ccttacaacg gctctcccgc
    14161 tgacgccgtc ccggactgat gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc
    14221 ggtcggggag ctgttggctg gctgg
//
