LOCUS       sequence-379610-       14262 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 14262)
  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 14262)
  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-VD1
FEATURES             Location/Qualifiers
     source          1..14262
                     /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)"
     CDS             join(3045..3047,3048..3050,3051..3563)
                     /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(3090..3111)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3351..3370)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     promoter        3877..3979
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             4010..4315
                     /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      4411..5118
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        6119..6464
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    6493..6547
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     primer_bind     6575..6596
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6651..6674
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             6684..6707
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6714..6737
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        6934..7036
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             7067..7372
                     /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      7461..7713
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(7764..7781)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7764..7780)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7773..7795)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    7983..8007
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             9308..9937
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             10366..11439
                     /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      11505..11699
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(11935..11957)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    12043..12183
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     12098..12117
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(12198..12215)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12369..12957)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12449..12468)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(13044..13838)
                     /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 tacaatggtg agcaagggcg
     3061 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     3121 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     3181 agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg
     3241 gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     3301 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     3361 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     3421 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3481 acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     3541 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
     3601 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     3661 ccaagctgag caaaggagcc agtggaggtg ggtcagagtg gtctctgtcc agtcctggcc
     3721 tcgtcggcca ttctcgacta agttggcagc atcacccgac gcactttgcg ccgaataaat
     3781 acctgtgacg gaagatcact tcgcagaata aataaatcct ggtgtccctg ttgataccgg
     3841 gaagccctgg gccaactttt ggcgaaaatg agacgttgat cggcacgtaa gaggttccaa
     3901 ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc
     3961 aggagctaag gaagctaaac ttttgctgac gagaacaggg actggtgaaa tgcagtttaa
     4021 ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat
     4081 tattgacacg cctgggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga
     4141 taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat
     4201 gaccaccgat atggccagtg tgccggtatc cgttatcggg gaagaagtgg ctgatctcag
     4261 ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc
     4321 aggctccctt atacacaggg ccagtctggc cacttgtggt ctgctgagac ctagttaatc
     4381 tagacccggg ggttaattaa ccctagagtc ctgctttaat gagatatgcg agacgcctat
     4441 gatcgcatga tatttgcttt caattctgtt gtgcacgttg taaaaaacct gagcatgtgt
     4501 agctcagatc cttaccgccg gtttcggttc attctaatga atatatcacc cgttactatc
     4561 gtatttttat gaataatatt ctccgttcaa tttactgatt gtaccctact acttatatgt
     4621 acaatattaa aatgaaaaca atatattgtg ctgaataggt ttatagcgac atctatgata
     4681 gagcgccaca ataacaaaca attgcgtttt attattacaa atccaatttt aaaaaaagcg
     4741 gcagaaccgg tcaaacctaa aagactgatt acataaatct tattcaaatt tcaaaagtgc
     4801 cccaggggct agtatctacg acacaccgag cggcgaacta ataacgctca ctgaagggaa
     4861 ctccggttcc ccgccggcgc gcatgggtga gattccttga agttgagtat tggccgtccg
     4921 ctctaccgaa agttacgggc accattcaac ccggtccagc acggcggccg ggtaaccgac
     4981 ttgctgcccc gagaattatg cagcattttt ttggtgtatg tgggccccaa atgaagtgca
     5041 ggtcaaacct tgacagtgac gacaaatcgt tgggcgggtc cagggcgaat tttgcgacaa
     5101 catgtcgagg ctcagcagga cctgctcata tgttccaatc ccacaaaaat ctgagcttaa
     5161 cagcacagtt gctcctctca gagcagaatc gggtattcaa caccctcata tcaactacta
     5221 cgttgtgtat aacggtccac atgccggtat atacgatgac tggggttgta caaaggcggc
     5281 aacaaacggc gttcccggag ttgcacacaa gaaatttgcc actattacag aggcaagagc
     5341 agcagctgac gcgtacacaa caagtcagca aacagacagg ttgaacttca tccccaaagg
     5401 agaagctcaa ctcaagccca agagctttgc taaggcccta acaagcccac caaagcaaaa
     5461 agcccactgg ctcacgctag gaaccaaaag gcccagcagt gatccagccc caaaagagat
     5521 ctcctttgcc ccggagatta caatggacga tttcctctat ctttacgatc taggaaggaa
     5581 gttcgaaggt gaaggtgacg acactatgtt caccactgat aatgagaagg ttagcctctt
     5641 caatttcaga aagaatgctg acccacagat ggttagagag gcctacgcag caggtctcat
     5701 caagacgatc tacccgagta acaatctcca ggagatcaaa taccttccca agaaggttaa
     5761 agatgcagtc aaaagattca ggactaattg catcaagaac acagagaaag acatatttct
     5821 caagatcaga agtactattc cagtatggac gattcaaggc ttgcttcata aaccaaggca
     5881 agtaatagag attggagtct ctaaaaaggt agttcctact gaatctaagg ccatgcatgg
     5941 agtctaagat tcaaatcgag gatctaacag aactcgccgt gaagactggc gaacagttca
     6001 tacagagtct tttacgactc aatgacaaga agaaaatctt cgtcaacatg gtggagcacg
     6061 acactctggt ctactccaaa aatgtcaaag atacagtctc agaagaccaa agggctattg
     6121 agacttttca acaaaggata atttcgggaa acctcctcgg attccattgc ccagctatct
     6181 gtcacttcat cgaaaggaca gtagaaaagg aaggtggctc ctacaaatgc catcattgcg
     6241 ataaaggaaa ggctatcatt caagatctct ctgccgacag tggtcccaaa gatggacccc
     6301 cacccacgag gagcatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg
     6361 attgatgtga catctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag
     6421 acccttcctc tatataagga agttcatttc atttggagag gacacgcctc aggtacagag
     6481 catattttta caacaattac caacaacaac aaacaacaaa caacattaca attacattta
     6541 caattacaat ggaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     6601 ccgccgggat cactctcggc atggacgagc tgtacaaggg atctgggtca tggtctcatc
     6661 cacaattcga gaagtcagga ggagattata aggatgatga tgataaggct tcttggtcac
     6721 atcctcagtt tgaaaagtct ggagcttcag gaagagttgg ctcgaccatg cgtggcctcg
     6781 tcggccattc tcgactaagt tggcagcatc acccgacgca ctttgcgccg aataaatacc
     6841 tgtgacggaa gatcacttcg cagaataaat aaatcctggt gtccctgttg ataccgggaa
     6901 gccctgggcc aacttttggc gaaaatgaga cgttgatcgg cacgtaagag gttccaactt
     6961 tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg
     7021 agctaaggaa gctaaacttt tgctgacgag aacagggact ggtgaaatgc agtttaaggt
     7081 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
     7141 tgacacgcct gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
     7201 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
     7261 caccgatatg gccagtgtgc cggtatccgt tatcggggaa gaagtggctg atctcagcca
     7321 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg
     7381 ctcccttata cacagggcca gtctggccag acctggatgt ttccagtgga cgtccggata
     7441 agcgatcgcc atttaaatag gatcgttcaa acatttggca ataaagtttc ttaagattga
     7501 atcctgttgc cggtcttgcg atgattatca tataatttct gttgaattac gttaagcatg
     7561 taataattaa catgtaatgc atgacgttat ttatgagatg ggtttttatg attagagtcc
     7621 cgcaattata catttaatac gcgatagaaa acaaaatata gcgcgcaaac taggataaat
     7681 tatcgcgcgc ggtgtcatct atgttactag atcgttaacg atatgtcgag gctcagcagg
     7741 acctgcaggc atgcaagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     7801 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     7861 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgc
     7921 tagagcagct tgagcttgga tcagattgtc gtttcccgcc ttcagtttaa actatcagtg
     7981 tttgacagga tatattggcg ggtaaaccta agagaaaaga gcgtttatta gaataatcgg
     8041 atatttaaaa gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc atgccaacca
     8101 cagggttccc ctcgggatca aagtactttg atccaacccc tccgctgcta tagtgcagtc
     8161 ggcttctgac gttcagtgca gccgtcttct gaaaacgaca tgtcgcacaa gtcctaagtt
     8221 acgcgacagg ctgccgccct gcccttttcc tggcgttttc ttgtcgcgtg ttttagtcgc
     8281 ataaagtaga atacttgcga ctagaaccgg agacattacg ccatgaacaa gagcgccgcc
     8341 gctggcctgc tgggctatgc ccgcgtcagc accgacgacc aggacttgac caaccaacgg
     8401 gccgaactgc acgcggccgg ctgcaccaag ctgttttccg agaagatcac cggcaccagg
     8461 cgcgaccgcc cggagctggc caggatgctt gaccacctac gccctggcga cgttgtgaca
     8521 gtgaccaggc tagaccgcct ggcccgcagc acccgcgacc tactggacat tgccgagcgc
     8581 atccaggagg ccggcgcggg cctgcgtagc ctggcagagc cgtgggccga caccaccacg
     8641 ccggccggcc gcatggtgtt gaccgtgttc gccggcattg ccgagttcga gcgttcccta
     8701 atcatcgacc gcacccggag cgggcgcgag gccgccaagg cgcgaggcgt gaagtttggc
     8761 ccccgcccta ccctcacccc ggcacagatc gcgcacgccc gcgagctgat cgaccaggaa
     8821 ggccgcaccg tgaaagaggc ggctgcactg cttggcgtgc atcgctcgac cctgtaccgc
     8881 gcacttgagc gcagcgagga agtgacgccc accgaggcca ggcggcgcgg tgccttccgt
     8941 gaggacgcat tgaccgaggc cgacgccctg gcggccgccg agaatgaacg ccaagaggaa
     9001 caagcatgaa accgcaccag gacggccagg acgaaccgtt tttcattacc gaagagatcg
     9061 aggcggagat gatcgcggcc gggtacgtgt tcgagccgcc cgcgcacgtc tcaaccgtgc
     9121 ggctgcatga aatcctggcc ggtttgtctg atgccaagct ggcggcctgg ccggcgagct
     9181 tggccgctga agaaaccgag cgccgccgtc taaaaaggtg atgtgtattt gagtaaaaca
     9241 gcttgcgtca tgcggtcgct gcgtatatga tgcgatgagt aaataaacaa atacgcaagg
     9301 ggaacgcatg aaggttatcg ctgtacttaa ccagaaaggc gggtcaggca agacgaccat
     9361 cgcaacccat ctagcccgcg ccctgcaact cgccggggcc gatgttctgt tagtcgattc
     9421 cgatccccag ggcagtgccc gcgattgggc ggccgtgcgg gaagatcaac cgctaaccgt
     9481 tgtcggcatc gaccgcccga cgattgaccg cgacgtgaag gccatcggcc ggcgcgactt
     9541 cgtagtgatc gacggagcgc cccaggcggc ggacttggct gtgtccgcga tcaaggcagc
     9601 cgacttcgtg ctgattccgg tgcagccaag cccttacgac atatgggcca ccgccgacct
     9661 ggtggagctg gttaagcagc gcattgaggt cacggatgga aggctacaag cggcctttgt
     9721 cgtgtcgcgg gcgatcaaag gcacgcgcat cggcggtgag gttgccgagg cgctggccgg
     9781 gtacgagctg cccattcttg agtcccgtat cacgcagcgc gtgagctacc caggcactgc
     9841 cgccgccggc acaaccgttc ttgaatcaga acccgagggc gacgctgccc gcgaggtcca
     9901 ggcgctggcc gctgaaatta aatcaaaact catttgagtt aatgaggtaa agagaaaatg
     9961 agcaaaagca caaacacgct aagtgccggc cgtccgagcg cacgcagcag caaggctgca
    10021 acgttggcca gcctggcaga cacgccagcc atgaagcggg tcaactttca gttgccggcg
    10081 gaggatcaca ccaagctgaa gatgtacgcg gtacgccaag gcaagaccat taccgagctg
    10141 ctatctgaat acatcgcgca gctaccagag taaatgagca aatgaataaa tgagtagatg
    10201 aattttagcg gctaaaggag gcggcatgga aaatcaagaa caaccaggca ccgacgccgt
    10261 ggaatgcccc atgtgtggag gaacgggcgg ttggccaggc gtaagcggct gggttgcctg
    10321 ccggccctgc aatggcactg gaacccccaa gcccgaggaa tcggcgtgag cggtcgcaaa
    10381 ccatccggcc cggtacaaat cggcgcggcg ctgggtgatg acctggtgga gaagttgaag
    10441 gcggcgcagg ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg tgaatcgtgg
    10501 caagcggccg ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc cggtgcgccg
    10561 tcgattagga agccgcccaa gggcgacgag caaccagatt ttttcgttcc gatgctctat
    10621 gacgtgggca cccgcgatag tcgcagcatc atggacgtgg ccgttttccg tctgtcgaag
    10681 cgtgaccgac gagctggcga ggtgatccgc tacgagcttc cagacgggca cgtagaggtt
    10741 tccgcagggc cggcgggcat ggccagtgtg tgggattacg acctggtact gatggcggtt
    10801 tcccatctaa ccgaatccat gaaccgatac cgggaaggga agggagacaa gcccggccgc
    10861 gtgttccgtc cacacgttgc ggacgtactc aagttctgcc ggcgagccga tggcggaaag
    10921 cagaaagacg acctggtaga aacctgcatt cggttaaaca ccacgcacgt tgccatgcag
    10981 cgtacgaaga aggccaagaa cggccgcctg gtgacggtat ccgagggtga agccttgatt
    11041 agccgctaca agatcgtaaa gagcgaaacc gggcggccgg agtacatcga gatcgagcta
    11101 gctgattgga tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct gacggttcac
    11161 cccgattact ttttgatcga tcccggcatc ggccgttttc tctaccgcct ggcacgccgc
    11221 gccgcaggca aggcagaagc cagatggttg ttcaagacga tctacgaacg cagtggcagc
    11281 gccggagagt tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc aaatgacctg
    11341 ccggagtacg atttgaagga ggaggcgggg caggctggcc cgatcctagt catgcgctac
    11401 cgcaacctga tcgagggcga agcatccgcc ggttcctaat gtacggagca gatgctaggg
    11461 caaattgccc tagcagggga aaaaggtcga aaaggtctct ttcctgtgga tagcacgtac
    11521 attgggaacc caaagccgta cattgggaac cggaacccgt acattgggaa cccaaagccg
    11581 tacattggga accggtcaca catgtaagtg actgatataa aagagaaaaa aggcgatttt
    11641 tccgcctaaa actctttaaa acttattaaa actcttaaaa cccgcctggc ctgtgcataa
    11701 ctgtctggcc agcgcacagc cgaagagctg caaaaagcgc ctacccttcg gtcgctgcgc
    11761 tccctacgcc ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc aaaaatggct
    11821 ggcctacggc caggcaatct accagggcgc ggacaagccg cgccgtcgcc actcgaccgc
    11881 cggcgcccac atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg aaaacctctg
    11941 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
    12001 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc
    12061 acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg
    12121 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
    12181 aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
    12241 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
    12301 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
    12361 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
    12421 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
    12481 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
    12541 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
    12601 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
    12661 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
    12721 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
    12781 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
    12841 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
    12901 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
    12961 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
    13021 attttggtca tgcattctag gtactaaaac aattcatcca gtaaaatata atattttatt
    13081 ttctcccaat caggcttgat ccccagtaag tcaaaaaata gctcgacata ctgttcttcc
    13141 ccgatatcct ccctgatcga ccggacgcag aaggcaatgt cataccactt gtccgccctg
    13201 ccgcttctcc caagatcaat aaagccactt actttgccat ctttcacaaa gatgttgctg
    13261 tctcccaggt cgccgtggga aaagacaagt tcctcttcgg gcttttccgt ctttaaaaaa
    13321 tcatacagct cgcgcggatc tttaaatgga gtgtcttctt cccagttttc gcaatccaca
    13381 tcggccagat cgttattcag taagtaatcc aattcggcta agcggctgtc taagctattc
    13441 gtatagggac aatccgatat gtcgatggag tgaaagagcc tgatgcactc cgcatacagc
    13501 tcgataatct tttcagggct ttgttcatct tcatactctt ccgagcaaag gacgccatcg
    13561 gcctcactca tgagcagatt gctccagcca tcatgccgtt caaagtgcag gacctttgga
    13621 acaggcagct ttccttccag ccatagcatc atgtcctttt cccgttccac atcataggtg
    13681 gtccctttat accggctgtc cgtcattttt aaatataggt tttcattttc tcccaccagc
    13741 ttatatacct tagcaggaga cattccttcc gtatctttta cgcagcggta tttttcgatc
    13801 agttttttca attccggtga tattctcatt ttagccattt attatttcct tcctcttttc
    13861 tacagtattt aaagataccc caagaagcta attataacaa gacgaactcc aattcactgt
    13921 tccttgcatt ctaaaacctt aaataccaga aaacagcttt ttcaaagttg ttttcaaagt
    13981 tggcgtataa catagtatcg acggagccga ttttgaaacc gcggtgatca caggcagcaa
    14041 cgctctgtca tcgttacaat caacatgcta ccctccgcga gatcatccgt gtttcaaacc
    14101 cggcagctta gttgccgttc ttccgaatag catcggtaac atgagcaaag tctgccgcct
    14161 tacaacggct ctcccgctga cgccgtcccg gactgatggg ctgcctgtat cgagtggtga
    14221 ttttgtgccg agctgccggt cggggagctg ttggctggct gg
//
