LOCUS       sequence-380043-       14255 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 14255)
  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 14255)
  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-VD2
FEATURES             Location/Qualifiers
     source          1..14255
                     /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)"
     promoter        6729..6831
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             6862..7167
                     /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             7246..7269
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             7279..7302
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7309..7332
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     7368..7389
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      7454..7706
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(7757..7774)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7757..7773)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7766..7788)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    7976..8000
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             9301..9930
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             10359..11432
                     /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      11498..11692
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(11928..11950)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    12036..12176
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     12091..12110
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(12191..12208)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12362..12950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12442..12461)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(13037..13831)
                     /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 caattacaga gttggctcga ccatgcgtgg cctcgtcggc cattctcgac taagttggca
     6601 gcatcacccg acgcactttg cgccgaataa atacctgtga cggaagatca cttcgcagaa
     6661 taaataaatc ctggtgtccc tgttgatacc gggaagccct gggccaactt ttggcgaaaa
     6721 tgagacgttg atcggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta
     6781 ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa acttttgctg
     6841 acgagaacag ggactggtga aatgcagttt aaggtttaca cctataaaag agagagccgt
     6901 tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcctgggcg acggatggtg
     6961 atccccctgg ccagtgcacg tctgctgtca gataaagtct cccgtgaact ttacccggtg
     7021 gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggta
     7081 tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc
     7141 attaacctga tgttctgggg aatataaatg tcaggctccc ttatacacag ggccagtctg
     7201 gccagacctg gatgtttcca gtggtccgga ggttctggtg gagcttggtc tcatccacaa
     7261 ttcgagaagt caggaggaga ttataaggat gatgatgata aggcttcttg gtcacatcct
     7321 cagtttgaaa agggatctgg gtcagacccc aacgagaagc gcgatcacat ggtcctgctg
     7381 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagcgatc
     7441 gccatttaaa taggatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt
     7501 tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc atgtaataat
     7561 taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt
     7621 atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg
     7681 cgcggtgtca tctatgttac tagatcgtta acgatatgtc gaggctcagc aggacctgca
     7741 ggcatgcaag cttggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     7801 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     7861 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgctagagca
     7921 gcttgagctt ggatcagatt gtcgtttccc gccttcagtt taaactatca gtgtttgaca
     7981 ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat cggatattta
     8041 aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgcatgccaa ccacagggtt
     8101 cccctcggga tcaaagtact ttgatccaac ccctccgctg ctatagtgca gtcggcttct
     8161 gacgttcagt gcagccgtct tctgaaaacg acatgtcgca caagtcctaa gttacgcgac
     8221 aggctgccgc cctgcccttt tcctggcgtt ttcttgtcgc gtgttttagt cgcataaagt
     8281 agaatacttg cgactagaac cggagacatt acgccatgaa caagagcgcc gccgctggcc
     8341 tgctgggcta tgcccgcgtc agcaccgacg accaggactt gaccaaccaa cgggccgaac
     8401 tgcacgcggc cggctgcacc aagctgtttt ccgagaagat caccggcacc aggcgcgacc
     8461 gcccggagct ggccaggatg cttgaccacc tacgccctgg cgacgttgtg acagtgacca
     8521 ggctagaccg cctggcccgc agcacccgcg acctactgga cattgccgag cgcatccagg
     8581 aggccggcgc gggcctgcgt agcctggcag agccgtgggc cgacaccacc acgccggccg
     8641 gccgcatggt gttgaccgtg ttcgccggca ttgccgagtt cgagcgttcc ctaatcatcg
     8701 accgcacccg gagcgggcgc gaggccgcca aggcgcgagg cgtgaagttt ggcccccgcc
     8761 ctaccctcac cccggcacag atcgcgcacg cccgcgagct gatcgaccag gaaggccgca
     8821 ccgtgaaaga ggcggctgca ctgcttggcg tgcatcgctc gaccctgtac cgcgcacttg
     8881 agcgcagcga ggaagtgacg cccaccgagg ccaggcggcg cggtgccttc cgtgaggacg
     8941 cattgaccga ggccgacgcc ctggcggccg ccgagaatga acgccaagag gaacaagcat
     9001 gaaaccgcac caggacggcc aggacgaacc gtttttcatt accgaagaga tcgaggcgga
     9061 gatgatcgcg gccgggtacg tgttcgagcc gcccgcgcac gtctcaaccg tgcggctgca
     9121 tgaaatcctg gccggtttgt ctgatgccaa gctggcggcc tggccggcga gcttggccgc
     9181 tgaagaaacc gagcgccgcc gtctaaaaag gtgatgtgta tttgagtaaa acagcttgcg
     9241 tcatgcggtc gctgcgtata tgatgcgatg agtaaataaa caaatacgca aggggaacgc
     9301 atgaaggtta tcgctgtact taaccagaaa ggcgggtcag gcaagacgac catcgcaacc
     9361 catctagccc gcgccctgca actcgccggg gccgatgttc tgttagtcga ttccgatccc
     9421 cagggcagtg cccgcgattg ggcggccgtg cgggaagatc aaccgctaac cgttgtcggc
     9481 atcgaccgcc cgacgattga ccgcgacgtg aaggccatcg gccggcgcga cttcgtagtg
     9541 atcgacggag cgccccaggc ggcggacttg gctgtgtccg cgatcaaggc agccgacttc
     9601 gtgctgattc cggtgcagcc aagcccttac gacatatggg ccaccgccga cctggtggag
     9661 ctggttaagc agcgcattga ggtcacggat ggaaggctac aagcggcctt tgtcgtgtcg
     9721 cgggcgatca aaggcacgcg catcggcggt gaggttgccg aggcgctggc cgggtacgag
     9781 ctgcccattc ttgagtcccg tatcacgcag cgcgtgagct acccaggcac tgccgccgcc
     9841 ggcacaaccg ttcttgaatc agaacccgag ggcgacgctg cccgcgaggt ccaggcgctg
     9901 gccgctgaaa ttaaatcaaa actcatttga gttaatgagg taaagagaaa atgagcaaaa
     9961 gcacaaacac gctaagtgcc ggccgtccga gcgcacgcag cagcaaggct gcaacgttgg
    10021 ccagcctggc agacacgcca gccatgaagc gggtcaactt tcagttgccg gcggaggatc
    10081 acaccaagct gaagatgtac gcggtacgcc aaggcaagac cattaccgag ctgctatctg
    10141 aatacatcgc gcagctacca gagtaaatga gcaaatgaat aaatgagtag atgaatttta
    10201 gcggctaaag gaggcggcat ggaaaatcaa gaacaaccag gcaccgacgc cgtggaatgc
    10261 cccatgtgtg gaggaacggg cggttggcca ggcgtaagcg gctgggttgc ctgccggccc
    10321 tgcaatggca ctggaacccc caagcccgag gaatcggcgt gagcggtcgc aaaccatccg
    10381 gcccggtaca aatcggcgcg gcgctgggtg atgacctggt ggagaagttg aaggcggcgc
    10441 aggccgccca gcggcaacgc atcgaggcag aagcacgccc cggtgaatcg tggcaagcgg
    10501 ccgctgatcg aatccgcaaa gaatcccggc aaccgccggc agccggtgcg ccgtcgatta
    10561 ggaagccgcc caagggcgac gagcaaccag attttttcgt tccgatgctc tatgacgtgg
    10621 gcacccgcga tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg aagcgtgacc
    10681 gacgagctgg cgaggtgatc cgctacgagc ttccagacgg gcacgtagag gtttccgcag
    10741 ggccggcggg catggccagt gtgtgggatt acgacctggt actgatggcg gtttcccatc
    10801 taaccgaatc catgaaccga taccgggaag ggaagggaga caagcccggc cgcgtgttcc
    10861 gtccacacgt tgcggacgta ctcaagttct gccggcgagc cgatggcgga aagcagaaag
    10921 acgacctggt agaaacctgc attcggttaa acaccacgca cgttgccatg cagcgtacga
    10981 agaaggccaa gaacggccgc ctggtgacgg tatccgaggg tgaagccttg attagccgct
    11041 acaagatcgt aaagagcgaa accgggcggc cggagtacat cgagatcgag ctagctgatt
    11101 ggatgtaccg cgagatcaca gaaggcaaga acccggacgt gctgacggtt caccccgatt
    11161 actttttgat cgatcccggc atcggccgtt ttctctaccg cctggcacgc cgcgccgcag
    11221 gcaaggcaga agccagatgg ttgttcaaga cgatctacga acgcagtggc agcgccggag
    11281 agttcaagaa gttctgtttc accgtgcgca agctgatcgg gtcaaatgac ctgccggagt
    11341 acgatttgaa ggaggaggcg gggcaggctg gcccgatcct agtcatgcgc taccgcaacc
    11401 tgatcgaggg cgaagcatcc gccggttcct aatgtacgga gcagatgcta gggcaaattg
    11461 ccctagcagg ggaaaaaggt cgaaaaggtc tctttcctgt ggatagcacg tacattggga
    11521 acccaaagcc gtacattggg aaccggaacc cgtacattgg gaacccaaag ccgtacattg
    11581 ggaaccggtc acacatgtaa gtgactgata taaaagagaa aaaaggcgat ttttccgcct
    11641 aaaactcttt aaaacttatt aaaactctta aaacccgcct ggcctgtgca taactgtctg
    11701 gccagcgcac agccgaagag ctgcaaaaag cgcctaccct tcggtcgctg cgctccctac
    11761 gccccgccgc ttcgcgtcgg cctatcgcgg ccgctggccg ctcaaaaatg gctggcctac
    11821 ggccaggcaa tctaccaggg cgcggacaag ccgcgccgtc gccactcgac cgccggcgcc
    11881 cacatcaagg caccctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
    11941 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
    12001 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
    12061 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
    12121 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
    12181 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
    12241 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
    12301 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
    12361 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
    12421 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
    12481 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
    12541 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
    12601 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
    12661 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
    12721 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
    12781 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
    12841 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
    12901 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
    12961 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
    13021 tcatgcattc taggtactaa aacaattcat ccagtaaaat ataatatttt attttctccc
    13081 aatcaggctt gatccccagt aagtcaaaaa atagctcgac atactgttct tccccgatat
    13141 cctccctgat cgaccggacg cagaaggcaa tgtcatacca cttgtccgcc ctgccgcttc
    13201 tcccaagatc aataaagcca cttactttgc catctttcac aaagatgttg ctgtctccca
    13261 ggtcgccgtg ggaaaagaca agttcctctt cgggcttttc cgtctttaaa aaatcataca
    13321 gctcgcgcgg atctttaaat ggagtgtctt cttcccagtt ttcgcaatcc acatcggcca
    13381 gatcgttatt cagtaagtaa tccaattcgg ctaagcggct gtctaagcta ttcgtatagg
    13441 gacaatccga tatgtcgatg gagtgaaaga gcctgatgca ctccgcatac agctcgataa
    13501 tcttttcagg gctttgttca tcttcatact cttccgagca aaggacgcca tcggcctcac
    13561 tcatgagcag attgctccag ccatcatgcc gttcaaagtg caggaccttt ggaacaggca
    13621 gctttccttc cagccatagc atcatgtcct tttcccgttc cacatcatag gtggtccctt
    13681 tataccggct gtccgtcatt tttaaatata ggttttcatt ttctcccacc agcttatata
    13741 ccttagcagg agacattcct tccgtatctt ttacgcagcg gtatttttcg atcagttttt
    13801 tcaattccgg tgatattctc attttagcca tttattattt ccttcctctt ttctacagta
    13861 tttaaagata ccccaagaag ctaattataa caagacgaac tccaattcac tgttccttgc
    13921 attctaaaac cttaaatacc agaaaacagc tttttcaaag ttgttttcaa agttggcgta
    13981 taacatagta tcgacggagc cgattttgaa accgcggtga tcacaggcag caacgctctg
    14041 tcatcgttac aatcaacatg ctaccctccg cgagatcatc cgtgtttcaa acccggcagc
    14101 ttagttgccg ttcttccgaa tagcatcggt aacatgagca aagtctgccg ccttacaacg
    14161 gctctcccgc tgacgccgtc ccggactgat gggctgcctg tatcgagtgg tgattttgtg
    14221 ccgagctgcc ggtcggggag ctgttggctg gctgg
//
