LOCUS       sequence-203657-       12931 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ratiometric BiFC analysis in plants (transient transformation).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12931)
  AUTHORS   Grefen C, Blatt MR
  TITLE     A 2in1 cloning system enables ratiometric bimolecular fluorescence 
            complementation (rBiFC).
  JOURNAL   Biotechniques. 2012 Oct 2;53(5):311-14. doi: 10.2144/000113941.
  PUBMED    23066669
REFERENCE   2  (bases 1 to 12931)
  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"; doi: "10"; journalName: 
            "Biotechniques"; date: "2012-10-2- 2"; volume: "53"; issue: "5"; 
            pages: "311-14"
COMMENT     Sequence Label: pBiFCt-2in1-CN
FEATURES             Location/Qualifiers
     source          1..12931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        93..437
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     protein_bind    532..655
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(690..695,696..713,714..720)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    728..744
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     733..755
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     752..768
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     752..768
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    complement(1029..1153)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1170..1196
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(1230..1232,1233..1235,1236..1694)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMA"
     primer_bind     complement(1275..1296)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1536..1555)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     promoter        2021..2365
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             2453..3127
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             2453..3127
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     2851..2870
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3045..3068
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     promoter        3482..3826
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     4105..4126
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4205..4234
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    4253..4377
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(4402..4407,4408..4425,4426..4432)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             4486..5145
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4552..4571)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             5487..5792
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     5671..5690
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(5833..5957)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    6281..6305
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             6826..7617
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      7863..8451
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8352..8371
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8605..8622
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8637..8777
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8703..8722)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8863..8885
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      9121..9315
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(9381..10454)
                     /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"
     CDS             complement(10883..11512)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    12843..12867
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
ORIGIN
        1 actagccctc tgcaggtcaa catggtggag cacgacactc tcgtctactc caagaatatc
       61 aaagatacag tctcagaaga ccagagggct attgagactt ttcaacaaag ggtaatatcg
      121 ggaaacctcc tcggattcca ttgcccagct atctgtcact tcatcgaaag gacagtagaa
      181 aaggaagatg gcttctacaa atgccatcat tgcgataaag gaaaggctat cgttcaagat
      241 gcctctaccg acagtggtcc caaagatgga cccccaccca cgaggaacat cgtggaaaaa
      301 gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgatatctc cactgacgta
      361 agggatgacg cacaatccca ctatccttcg caagaccctt cctctatata aggaagttca
      421 tttcatttgg agaggacctc gagcgcgcgt atttttaaac aattaccaac aacaacaaca
      481 aacaacaaca acattacatt ttacattcta caactacatc tagtcaattg gcaactttgt
      541 ataataaagt tgaacgagaa acgtaaaatg atataaatat caatatatta aattagattt
      601 tgcataaaaa acagactaca taatactgta aaacacaaca tatccagtca ctatggcggc
      661 cgcgggctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
      721 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacccc
      781 gtccctgcac gcctgtcgtt caaccctgga agatccgcgt gtcccgagtt ctaacagtct
      841 ggccgtcgtt ctgcagcgtc gcgattggga aaacccgggc gtgacccaac tgaatcgtct
      901 ggcggcccat ccgccgtttg cgtcgtggcg taactcagaa gaagcccgta ccgaccgtcc
      961 gtctcaacaa ctgcgctcct aacactataa aaagctgatg cagacggggc ttttttctgc
     1021 aggtcgacca tagtgactgg atatgttgtg ttttacagta ttatgtagtc tgttttttat
     1081 gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt tcagctttct
     1141 tgtacaaagt ggtgtctagg ttaaacatgt atccttatga tgttcctgat tatgctacta
     1201 gtggaggtgg atctggaggt ggaagtagaa tggtgagcaa gggcgaggag ctgttcaccg
     1261 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     1321 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     1381 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgcagt
     1441 gcttcgccag gtaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1501 aaggctacgt ccaggagaga accatcttct tcaaggacga cggcaactac aagaccaggg
     1561 ccgaggtgaa gttcgagggc gacaccctgg tgaacagaat cgagctgaag ggcatcgact
     1621 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1681 tctatatcat ggcctgatta taagtaagcg gccatgctag agtccgcaaa aatcaccagt
     1741 ctctctctac aaatctatct ctctctattt ttctccagaa taatgtgtga gtagttccca
     1801 gataagggaa ttagggttct tatagggttt cgctcatgtg ttgagcatat aagaaaccct
     1861 tagtatgtat ttgtatttgt aaaatacttc tatcaataaa atttctaatt cctaaaacca
     1921 aaatccagtg acctaggagg gctgtcaaca tggtggagca cgacactctc gtctactcca
     1981 agaatatcaa agatacagtc tcagaagacc agagggctat tgagactttt caacaaaggg
     2041 taatatcggg aaacctcctc ggattccatt gcccagctat ctgtcacttc atcgaaagga
     2101 cagtagaaaa ggaagatggc ttctacaaat gccatcattg cgataaagga aaggctatcg
     2161 ttcaagatgc ctctaccgac agtggtccca aagatggacc cccacccacg aggaacatcg
     2221 tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt gatatctcca
     2281 ctgacgtaag ggatgacgca caatcccact atccttcgca agacccttcc tctatataag
     2341 gaagttcatt tcatttggag aggacctcga gcgcgcgtat ttttaaacaa ttaccaacaa
     2401 caacaacaaa caacaacaac attacatttt acattctaca actacatcta gtatggcctc
     2461 ctccgaggac gtcatcaagg agttcatgcg cttcaaggtg cgcatggagg gctccgtgaa
     2521 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     2581 cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc
     2641 tcagttccag tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
     2701 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     2761 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
     2821 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
     2881 ggaggcctcc accgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagat
     2941 gaggctgaag ctgaaggacg gcggccacta cgacgccgag gtcaagacca cctacatggc
     3001 caagaagccc gtgcagctgc ccggcgccta caagaccgac atcaagctgg acatcacctc
     3061 ccacaacgag gactacacca tcgtggaaca gtacgagcgc gccgagggcc gccactccac
     3121 cggcgcccaa ttgttaatca caagtttgta caaagtggtg attaagcggc catgctagag
     3181 tccgcaaaaa tcaccagtct ctctctacaa atctatctct ctctattttt ctccagaata
     3241 atgtgtgagt agttcccaga taagggaatt agggttctta tagggtttcg ctcatgtgtt
     3301 gagcatataa gaaaccctta gtatgtattt gtatttgtaa aatacttcta tcaataaaat
     3361 ttctaattcc taaaaccaaa atccagtgac cttaccctct gcaggtcaac atggtggagc
     3421 acgacactct cgtctactcc aagaatatca aagatacagt ctcagaagac cagagggcta
     3481 ttgagacttt tcaacaaagg gtaatatcgg gaaacctcct cggattccat tgcccagcta
     3541 tctgtcactt catcgaaagg acagtagaaa aggaagatgg cttctacaaa tgccatcatt
     3601 gcgataaagg aaaggctatc gttcaagatg cctctaccga cagtggtccc aaagatggac
     3661 ccccacccac gaggaacatc gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag
     3721 tggattgatg tgatatctcc actgacgtaa gggatgacgc acaatcccac tatccttcgc
     3781 aagacccttc ctctatataa ggaagttcat ttcatttgga gaggacctcg agcgcgcgta
     3841 tttttaaaca attaccaaca acaacaacaa acaacaacaa cattacattt tacattctac
     3901 aactacatct aggatggaca agcagaagaa cggcatcaag gtgaacttca agatcaggca
     3961 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     4021 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa
     4081 agaccccaac gagaagagag atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     4141 cactctcggc atggacgagc tgtacaagtc tggaggtgga tctggaggtg gaagtaggcc
     4201 aatggaacag aagcttatct cagaggagga cctggccggc actagtccta gaacaagttt
     4261 gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat
     4321 tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt cactatggcg
     4381 gccgcattag gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt
     4441 ttgagttagg atccgtcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
     4501 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     4561 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     4621 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
     4681 ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg
     4741 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
     4801 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
     4861 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
     4921 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     4981 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     5041 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
     5101 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaaagatc tggatccggc
     5161 ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata
     5221 tatactgata tgtatacccg aagtatgtca aaaagaggta tgctatgaag cagcgtatta
     5281 cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc
     5341 cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa
     5401 agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt
     5461 tgctgacgag aacaggggct ggtgaaatgc agtttaaggt ttacacctat aaaagagaga
     5521 gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga
     5581 tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc
     5641 cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc
     5701 cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa
     5761 acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt
     5821 ctgcaggtcg accatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt
     5881 ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact
     5941 tttctataca aagttggtca attgttataa gcggccatgc tagagtccgc aaaaatcacc
     6001 agtctctctc tacaaatcta tctctctcta tttttctcca gaataatgtg tgagtagttc
     6061 ccagataagg gaattagggt tcttataggg tttcgctcat gtgttgagca tataagaaac
     6121 ccttagtatg tatttgtatt tgtaaaatac ttctatcaat aaaatttcta attcctaaaa
     6181 ccaaaatcca gtgacctagg aggtaagtcc tcaattcggc gttaattcag tacattaaaa
     6241 acgtccgcaa tgtgttatta agttgtctaa gcgtcaattt gtttacacca caatatatcc
     6301 tgccaccagc cagccaacag ctccccgacc ggcagctcgg cacaaaatca ccactcgata
     6361 caggcagccc atcagtccgg gacggcgtca gcgggagagc cgttgtaagg cggcagactt
     6421 tgctcatgtt accgatgcta ttcggaagaa cggcaactaa gctgccgggt ttgaaacacg
     6481 gatgatctcg cggagggtag catgttgatt gtaacgatga cagagcgttg ctgcctgtga
     6541 tcaattcggg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag
     6601 tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc
     6661 gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc tatgcctcgg
     6721 gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac
     6781 gatgttacgc agcagggcag tcgccctaaa acaaagttaa acatcatggg ggaagcggtg
     6841 atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa
     6901 ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac
     6961 agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct
     7021 ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct
     7081 gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc
     7141 gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc
     7201 acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg
     7261 gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg
     7321 ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat
     7381 gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag
     7441 gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt
     7501 gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag
     7561 ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataatgt
     7621 ctagctagaa attcgttcaa gccgacgccg cttcgcggcg cggcttaact caagcgttag
     7681 atgcactaag cacataattg ctcacagcca aactatcagg tcaagtctgc ttttattatt
     7741 tttaagcgtg cataataagc cctacacaaa ttgggagata tatcatgcat gaccaaaatc
     7801 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     7861 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     7921 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     7981 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     8041 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     8101 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     8161 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     8221 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     8281 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     8341 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     8401 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     8461 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     8521 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     8581 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     8641 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     8701 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     8761 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     8821 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     8881 agaggttttc accgtcatca ccgaaacgcg cgaggcaggg tgccttgatg tgggcgccgg
     8941 cggtcgagtg gcgacggcgc ggcttgtccg cgccctggta gattgcctgg ccctaggcca
     9001 gccatttttg agcggccagc ggccgcgata ggccgacgcg aagcggcggg gcgtagggag
     9061 cgcagcgacc gaagggtagg cgctttttgc agctcttcgg ctgtgcgctg gccagacagt
     9121 tatgcacagg ccaggcgggt tttaagagtt ttaataagtt ttaaagagtt ttaggcggaa
     9181 aaatcgcctt ttttctcttt tatatcagtc acttacatgt gtgaccggtt cccaatgtac
     9241 ggctttgggt tcccaatgta cgggttccgg ttcccaatgt acggctttgg gttcccaatg
     9301 tacgtgctat ccacaggaaa gagacctttt cgaccttttt cccctgctag ggcaatttgc
     9361 cctagcatct gctccgtaca ttaggaaccg gcggatgctt cgccctcgat caggttgcgg
     9421 tagcgcatga ctaggatcgg gccagcctgc cccgcctcct ccttcaaatc gtactccggc
     9481 aggtcatttg acccgatcag cttgcgcacg gtgaaacaga acttcttgaa ctctccggcg
     9541 ctgccactgc gttcgtagat cgtcttgaac aaccatctgg cttctgcctt gcctgcggcg
     9601 cggcgtgcca ggcggtagag aaaacggccg atgccgggat cgatcaaaaa gtaatcgggg
     9661 tgaaccgtca gcacgtccgg gttcttgcct tctgtgatct cgcggtacat ccaatcagct
     9721 agctcgatct cgatgtactc cggccgcccg gtttcgctct ttacgatctt gtagcggcta
     9781 atcaaggctt caccctcgga taccgtcacc aggcggccgt tcttggcctt cttcgtacgc
     9841 tgcatggcaa cgtgcgtggt gtttaaccga atgcaggttt ctaccaggtc gtctttctgc
     9901 tttccgccat cggctcgccg gcagaacttg agtacgtccg caacgtgtgg acggaacacg
     9961 cggccgggct tgtctccctt cccttcccgg tatcggttca tggattcggt tagatgggaa
    10021 accgccatca gtaccaggtc gtaatcccac acactggcca tgccggccgg ccctgcggaa
    10081 acctctacgt gcccgtctgg aagctcgtag cggatcacct cgccagctcg tcggtcacgc
    10141 ttcgacagac ggaaaacggc cacgtccatg atgctgcgac tatcgcgggt gcccacgtca
    10201 tagagcatcg gaacgaaaaa atctggttgc tcgtcgccct tgggcggctt cctaatcgac
    10261 ggcgcaccgg ctgccggcgg ttgccgggat tctttgcgga ttcgatcagc ggccgcttgc
    10321 cacgattcac cggggcgtgc ttctgcctcg atgcgttgcc gctgggcggc ctgcgcggcc
    10381 ttcaacttct ccaccaggtc atcacccagc gccgcgccga tttgtaccgg gccggatggt
    10441 ttgcgaccgc tcacgccgat tcctcgggct tgggggttcc agtgccattg cagggccggc
    10501 agacaaccca gccgcttacg cctggccaac cgcccgttcc tccacacatg gggcattcca
    10561 cggcgtcggt gcctggttgt tcttgatttt ccatgccgcc tcctttagcc gctaaaattc
    10621 atctactcat ttattcattt gctcatttac tctggtagct gcgcgatgta ttcagatagc
    10681 agctcggtaa tggtcttgcc ttggcgtacc gcgtacatct tcagcttggt gtgatcctcc
    10741 gccggcaact gaaagttgac ccgcttcatg gctggcgtgt ctgccaggct ggccaacgtt
    10801 gcagccttgc tgctgcgtgc gctcggacgg ccggcactta gcgtgtttgt gcttttgctc
    10861 attttctctt tacctcatta actcaaatga gttttgattt aatttcagcg gccagcgcct
    10921 ggacctcgcg ggcagcgtcg ccctcgggtt ctgattcaag aacggttgtg ccggcggcgg
    10981 cagtgcctgg gtagctcacg cgctgcgtga tacgggactc aagaatgggc agctcgtacc
    11041 cggccagcgc ctcggcaacc tcaccgccga tgcgcgtgcc tttgatcgcc cgcgacacga
    11101 caaaggccgc ttgtagcctt ccatccgtga cctcaatgcg ctgcttaacc agctccacca
    11161 ggtcggcggt ggcccatatg tcgtaagggc ttggctgcac cggaatcagc acgaagtcgg
    11221 ctgccttgat cgcggacaca gccaagtccg ccgcctgggg cgctccgtcg atcactacga
    11281 agtcgcgccg gccgatggcc ttcacgtcgc ggtcaatcgt cgggcggtcg atgccgacaa
    11341 cggttagcgg ttgatcttcc cgcacggccg cccaatcgcg ggcactgccc tggggatcgg
    11401 aatcgactaa cagaacatcg gccccggcga gttgcagggc gcgggctaga tgggttgcga
    11461 tggtcgtctt gcctgacccg cctttctggt taagtacagc gataaccttc atgcgttccc
    11521 cttgcgtatt tgtttattta ctcatcgcat catatacgca gcgaccgcat gacgcaagct
    11581 gttttactca aatacacatc acctttttag acggcggcgc tcggtttctt cagcggccaa
    11641 gctggccggc caggccgcca gcttggcatc agacaaaccg gccaggattt catgcagccg
    11701 cacggttgag acgtgcgcgg gcggctcgaa cacgtacccg gccgcgatca tctccgcctc
    11761 gatctcttcg gtaatgaaaa acggttcgtc ctggccgtcc tggtgcggtt tcatgcttgt
    11821 tcctcttggc gttcattctc ggcggccgcc agggcgtcgg cctcggtcaa tgcgtcctca
    11881 cggaaggcac cgcgccgcct ggcctcggtg ggcgtcactt cctcgctgcg ctcaagtgcg
    11941 cggtacaggg tcgagcgatg cacgccaagc agtgcagccg cctctttcac ggtgcggcct
    12001 tcctggtcga tcagctcgcg ggcgtgcgcg atctgtgccg gggtgagggt agggcggggg
    12061 ccaaacttca cgcctcgggc cttggcggcc tcgcgcccgc tccgggtgcg gtcgatgatt
    12121 agggaacgct cgaactcggc aatgccggcg aacacggtca acaccatgcg gccggccggc
    12181 gtggtggtgt cggcccacgg ctctgccagg ctacgcaggc ccgcgccggc ctcctggatg
    12241 cgctcggcaa tgtccagtag gtcgcgggtg ctgcgggcca ggcggtctag cctggtcact
    12301 gtcacaacgt cgccagggcg taggtggtca agcatcctgg ccagctccgg gcggtcgcgc
    12361 ctggtgccgg tgatcttctc ggaaaacagc ttggtgcagc cggccgcgtg cagttcggcc
    12421 cgttggttgg tcaagtcctg gtcgtcggtg ctgacgcggg catagcccag caggccagcg
    12481 gcggcgctct tgttcatggc gtaatgtctc cggttctagt cgcaagtatt ctactttatg
    12541 cgactaaaac acgcgacaag aaaacgccag gaaaagggca gggcggcagc ctgtcgcgta
    12601 acttaggact tgtgcgacat gtcgttttca gaagacggct gcactgaacg tcagaagccg
    12661 actgcactat agcagcggag gggttggatc aaagtacttt aaagtacttt aaagtacttt
    12721 aaagtacttt gatcccgagg ggaaccctgt ggttggcatg cacatacaaa tggacgaacg
    12781 gataaacctt ttcacgccct tttaaatatc cgattattct aataaacgct cttttctctt
    12841 aggtttaccc gccaatatat cctgtcaaac actgatagtt taaactgaag gcgggaaacg
    12901 acaatctgat ccaagctcaa gctaagctag g
//
