LOCUS       sequence-203654-       12925 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 12925)
  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 12925)
  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-NC
FEATURES             Location/Qualifiers
     source          1..12925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        93..437
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             join(531..533,534..536,537..995)
                     /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(576..597)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(837..856)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1029..1055
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    1066..1189
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1224..1229,1230..1247,1248..1254)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1262..1278
                     /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     1267..1289
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1286..1302
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1286..1302
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    complement(1563..1687)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2014..2358
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             2446..3120
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             2446..3120
                     /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     2844..2863
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3038..3061
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     promoter        3475..3819
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     protein_bind    3913..4037
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(4062..4067,4068..4085,4086..4092)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             4146..4805
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4212..4231)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             5147..5452
                     /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     5331..5350
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(5493..5617)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             5634..5663
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     5891..5912
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6275..6299
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             6820..7611
                     /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      7857..8445
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8346..8365
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8599..8616
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8631..8771
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8697..8716)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8857..8879
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      9115..9309
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(9375..10448)
                     /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(10877..11506)
                     /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    12837..12861
                     /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 atggtgagca
      541 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      601 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      661 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      721 ccttcggcta cggcctgcag tgcttcgcca ggtaccccga ccacatgaag cagcacgact
      781 tcttcaagtc cgccatgccc gaaggctacg tccaggagag aaccatcttc ttcaaggacg
      841 acggcaacta caagaccagg gccgaggtga agttcgaggg cgacaccctg gtgaacagaa
      901 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
      961 acaactacaa cagccacaac gtctatatca tggccggagg tggatctgga ggtggaagtc
     1021 agaacatgta tccttatgat gttcctgatt atgctagtca attggcaact ttgtataata
     1081 aagttgaacg agaaacgtaa aatgatataa atatcaatat attaaattag attttgcata
     1141 aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgg cggccgcggg
     1201 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     1261 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta ccccgtccct
     1321 gcacgcctgt cgttcaaccc tggaagatcc gcgtgtcccg agttctaaca gtctggccgt
     1381 cgttctgcag cgtcgcgatt gggaaaaccc gggcgtgacc caactgaatc gtctggcggc
     1441 ccatccgccg tttgcgtcgt ggcgtaactc agaagaagcc cgtaccgacc gtccgtctca
     1501 acaactgcgc tcctaacact ataaaaagct gatgcagacg gggctttttt ctgcaggtcg
     1561 accatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa
     1621 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcagct ttcttgtaca
     1681 aagtggtgtc taggttataa gcggccatgc tagagtccgc aaaaatcacc agtctctctc
     1741 tacaaatcta tctctctcta tttttctcca gaataatgtg tgagtagttc ccagataagg
     1801 gaattagggt tcttataggg tttcgctcat gtgttgagca tataagaaac ccttagtatg
     1861 tatttgtatt tgtaaaatac ttctatcaat aaaatttcta attcctaaaa ccaaaatcca
     1921 gtgacctagg agggctgtca acatggtgga gcacgacact ctcgtctact ccaagaatat
     1981 caaagataca gtctcagaag accagagggc tattgagact tttcaacaaa gggtaatatc
     2041 gggaaacctc ctcggattcc attgcccagc tatctgtcac ttcatcgaaa ggacagtaga
     2101 aaaggaagat ggcttctaca aatgccatca ttgcgataaa ggaaaggcta tcgttcaaga
     2161 tgcctctacc gacagtggtc ccaaagatgg acccccaccc acgaggaaca tcgtggaaaa
     2221 agaagacgtt ccaaccacgt cttcaaagca agtggattga tgtgatatct ccactgacgt
     2281 aagggatgac gcacaatccc actatccttc gcaagaccct tcctctatat aaggaagttc
     2341 atttcatttg gagaggacct cgagcgcgcg tatttttaaa caattaccaa caacaacaac
     2401 aaacaacaac aacattacat tttacattct acaactacat ctagtatggc ctcctccgag
     2461 gacgtcatca aggagttcat gcgcttcaag gtgcgcatgg agggctccgt gaacggccac
     2521 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     2581 ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     2641 cagtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
     2701 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     2761 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     2821 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     2881 tccaccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gatgaggctg
     2941 aagctgaagg acggcggcca ctacgacgcc gaggtcaaga ccacctacat ggccaagaag
     3001 cccgtgcagc tgcccggcgc ctacaagacc gacatcaagc tggacatcac ctcccacaac
     3061 gaggactaca ccatcgtgga acagtacgag cgcgccgagg gccgccactc caccggcgcc
     3121 caattgttaa tcacaagttt gtacaaagtg gtgattaagc ggccatgcta gagtccgcaa
     3181 aaatcaccag tctctctcta caaatctatc tctctctatt tttctccaga ataatgtgtg
     3241 agtagttccc agataaggga attagggttc ttatagggtt tcgctcatgt gttgagcata
     3301 taagaaaccc ttagtatgta tttgtatttg taaaatactt ctatcaataa aatttctaat
     3361 tcctaaaacc aaaatccagt gaccttaccc tctgcaggtc aacatggtgg agcacgacac
     3421 tctcgtctac tccaagaata tcaaagatac agtctcagaa gaccagaggg ctattgagac
     3481 ttttcaacaa agggtaatat cgggaaacct cctcggattc cattgcccag ctatctgtca
     3541 cttcatcgaa aggacagtag aaaaggaaga tggcttctac aaatgccatc attgcgataa
     3601 aggaaaggct atcgttcaag atgcctctac cgacagtggt cccaaagatg gacccccacc
     3661 cacgaggaac atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc aagtggattg
     3721 atgtgatatc tccactgacg taagggatga cgcacaatcc cactatcctt cgcaagaccc
     3781 ttcctctata taaggaagtt catttcattt ggagaggacc tcgagcgcgc gtatttttaa
     3841 acaattacca acaacaacaa caaacaacaa caacattaca ttttacattc tacaactaca
     3901 tctagtccta gaacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat
     3961 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
     4021 catatccagt cactatggcg gccgcattag gcaccccagg ctttacactt tatgcttccg
     4081 gctcgtataa tgtgtggatt ttgagttagg atccgtcgag attttcagga gctaaggaag
     4141 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     4201 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     4261 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     4321 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag
     4381 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     4441 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     4501 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     4561 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     4621 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     4681 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     4741 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     4801 cgtaaagatc tggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg
     4861 atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggta
     4921 tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc
     4981 atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt
     5041 ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt
     5101 attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc agtttaaggt
     5161 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
     5221 tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
     5281 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
     5341 caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca
     5401 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg
     5461 ctcccttata cacagccagt ctgcaggtcg accatagtga ctggatatgt tgtgttttac
     5521 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat
     5581 tttacgtttc tcgttcaact tttctataca aagttggtca attgttacca atggaacaga
     5641 agcttatctc agaggaggac ctggccggca ctagtggagg tggatctgga ggtggaagta
     5701 tggacaagca gaagaacggc atcaaggtga acttcaagat caggcacaac atcgaggacg
     5761 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     5821 tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga
     5881 agagagatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     5941 acgagctgta caagtagtta taagcggcca tgctagagtc cgcaaaaatc accagtctct
     6001 ctctacaaat ctatctctct ctatttttct ccagaataat gtgtgagtag ttcccagata
     6061 agggaattag ggttcttata gggtttcgct catgtgttga gcatataaga aacccttagt
     6121 atgtatttgt atttgtaaaa tacttctatc aataaaattt ctaattccta aaaccaaaat
     6181 ccagtgacct aggagggtaa gtcctcaatt cggcgttaat tcagtacatt aaaaacgtcc
     6241 gcaatgtgtt attaagttgt ctaagcgtca atttgtttac accacaatat atcctgccac
     6301 cagccagcca acagctcccc gaccggcagc tcggcacaaa atcaccactc gatacaggca
     6361 gcccatcagt ccgggacggc gtcagcggga gagccgttgt aaggcggcag actttgctca
     6421 tgttaccgat gctattcgga agaacggcaa ctaagctgcc gggtttgaaa cacggatgat
     6481 ctcgcggagg gtagcatgtt gattgtaacg atgacagagc gttgctgcct gtgatcaatt
     6541 cgggcacgaa cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt
     6601 atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg
     6661 gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc
     6721 aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt
     6781 acgcagcagg gcagtcgccc taaaacaaag ttaaacatca tgggggaagc ggtgatcgcc
     6841 gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg
     6901 ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat
     6961 attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc
     7021 aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa
     7081 gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg
     7141 caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc
     7201 gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt
     7261 ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat
     7321 gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg
     7381 cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc
     7441 gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct
     7501 agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa
     7561 gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata atgtctagct
     7621 agaaattcgt tcaagccgac gccgcttcgc ggcgcggctt aactcaagcg ttagatgcac
     7681 taagcacata attgctcaca gccaaactat caggtcaagt ctgcttttat tatttttaag
     7741 cgtgcataat aagccctaca caaattggga gatatatcat gcatgaccaa aatcccttaa
     7801 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7861 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7921 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7981 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     8041 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     8101 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     8161 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     8221 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     8281 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     8341 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     8401 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     8461 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     8521 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     8581 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     8641 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     8701 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     8761 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     8821 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     8881 tttcaccgtc atcaccgaaa cgcgcgaggc agggtgcctt gatgtgggcg ccggcggtcg
     8941 agtggcgacg gcgcggcttg tccgcgccct ggtagattgc ctggccctag gccagccatt
     9001 tttgagcggc cagcggccgc gataggccga cgcgaagcgg cggggcgtag ggagcgcagc
     9061 gaccgaaggg taggcgcttt ttgcagctct tcggctgtgc gctggccaga cagttatgca
     9121 caggccaggc gggttttaag agttttaata agttttaaag agttttaggc ggaaaaatcg
     9181 ccttttttct cttttatatc agtcacttac atgtgtgacc ggttcccaat gtacggcttt
     9241 gggttcccaa tgtacgggtt ccggttccca atgtacggct ttgggttccc aatgtacgtg
     9301 ctatccacag gaaagagacc ttttcgacct ttttcccctg ctagggcaat ttgccctagc
     9361 atctgctccg tacattagga accggcggat gcttcgccct cgatcaggtt gcggtagcgc
     9421 atgactagga tcgggccagc ctgccccgcc tcctccttca aatcgtactc cggcaggtca
     9481 tttgacccga tcagcttgcg cacggtgaaa cagaacttct tgaactctcc ggcgctgcca
     9541 ctgcgttcgt agatcgtctt gaacaaccat ctggcttctg ccttgcctgc ggcgcggcgt
     9601 gccaggcggt agagaaaacg gccgatgccg ggatcgatca aaaagtaatc ggggtgaacc
     9661 gtcagcacgt ccgggttctt gccttctgtg atctcgcggt acatccaatc agctagctcg
     9721 atctcgatgt actccggccg cccggtttcg ctctttacga tcttgtagcg gctaatcaag
     9781 gcttcaccct cggataccgt caccaggcgg ccgttcttgg ccttcttcgt acgctgcatg
     9841 gcaacgtgcg tggtgtttaa ccgaatgcag gtttctacca ggtcgtcttt ctgctttccg
     9901 ccatcggctc gccggcagaa cttgagtacg tccgcaacgt gtggacggaa cacgcggccg
     9961 ggcttgtctc ccttcccttc ccggtatcgg ttcatggatt cggttagatg ggaaaccgcc
    10021 atcagtacca ggtcgtaatc ccacacactg gccatgccgg ccggccctgc ggaaacctct
    10081 acgtgcccgt ctggaagctc gtagcggatc acctcgccag ctcgtcggtc acgcttcgac
    10141 agacggaaaa cggccacgtc catgatgctg cgactatcgc gggtgcccac gtcatagagc
    10201 atcggaacga aaaaatctgg ttgctcgtcg cccttgggcg gcttcctaat cgacggcgca
    10261 ccggctgccg gcggttgccg ggattctttg cggattcgat cagcggccgc ttgccacgat
    10321 tcaccggggc gtgcttctgc ctcgatgcgt tgccgctggg cggcctgcgc ggccttcaac
    10381 ttctccacca ggtcatcacc cagcgccgcg ccgatttgta ccgggccgga tggtttgcga
    10441 ccgctcacgc cgattcctcg ggcttggggg ttccagtgcc attgcagggc cggcagacaa
    10501 cccagccgct tacgcctggc caaccgcccg ttcctccaca catggggcat tccacggcgt
    10561 cggtgcctgg ttgttcttga ttttccatgc cgcctccttt agccgctaaa attcatctac
    10621 tcatttattc atttgctcat ttactctggt agctgcgcga tgtattcaga tagcagctcg
    10681 gtaatggtct tgccttggcg taccgcgtac atcttcagct tggtgtgatc ctccgccggc
    10741 aactgaaagt tgacccgctt catggctggc gtgtctgcca ggctggccaa cgttgcagcc
    10801 ttgctgctgc gtgcgctcgg acggccggca cttagcgtgt ttgtgctttt gctcattttc
    10861 tctttacctc attaactcaa atgagttttg atttaatttc agcggccagc gcctggacct
    10921 cgcgggcagc gtcgccctcg ggttctgatt caagaacggt tgtgccggcg gcggcagtgc
    10981 ctgggtagct cacgcgctgc gtgatacggg actcaagaat gggcagctcg tacccggcca
    11041 gcgcctcggc aacctcaccg ccgatgcgcg tgcctttgat cgcccgcgac acgacaaagg
    11101 ccgcttgtag ccttccatcc gtgacctcaa tgcgctgctt aaccagctcc accaggtcgg
    11161 cggtggccca tatgtcgtaa gggcttggct gcaccggaat cagcacgaag tcggctgcct
    11221 tgatcgcgga cacagccaag tccgccgcct ggggcgctcc gtcgatcact acgaagtcgc
    11281 gccggccgat ggccttcacg tcgcggtcaa tcgtcgggcg gtcgatgccg acaacggtta
    11341 gcggttgatc ttcccgcacg gccgcccaat cgcgggcact gccctgggga tcggaatcga
    11401 ctaacagaac atcggccccg gcgagttgca gggcgcgggc tagatgggtt gcgatggtcg
    11461 tcttgcctga cccgcctttc tggttaagta cagcgataac cttcatgcgt tccccttgcg
    11521 tatttgttta tttactcatc gcatcatata cgcagcgacc gcatgacgca agctgtttta
    11581 ctcaaataca catcaccttt ttagacggcg gcgctcggtt tcttcagcgg ccaagctggc
    11641 cggccaggcc gccagcttgg catcagacaa accggccagg atttcatgca gccgcacggt
    11701 tgagacgtgc gcgggcggct cgaacacgta cccggccgcg atcatctccg cctcgatctc
    11761 ttcggtaatg aaaaacggtt cgtcctggcc gtcctggtgc ggtttcatgc ttgttcctct
    11821 tggcgttcat tctcggcggc cgccagggcg tcggcctcgg tcaatgcgtc ctcacggaag
    11881 gcaccgcgcc gcctggcctc ggtgggcgtc acttcctcgc tgcgctcaag tgcgcggtac
    11941 agggtcgagc gatgcacgcc aagcagtgca gccgcctctt tcacggtgcg gccttcctgg
    12001 tcgatcagct cgcgggcgtg cgcgatctgt gccggggtga gggtagggcg ggggccaaac
    12061 ttcacgcctc gggccttggc ggcctcgcgc ccgctccggg tgcggtcgat gattagggaa
    12121 cgctcgaact cggcaatgcc ggcgaacacg gtcaacacca tgcggccggc cggcgtggtg
    12181 gtgtcggccc acggctctgc caggctacgc aggcccgcgc cggcctcctg gatgcgctcg
    12241 gcaatgtcca gtaggtcgcg ggtgctgcgg gccaggcggt ctagcctggt cactgtcaca
    12301 acgtcgccag ggcgtaggtg gtcaagcatc ctggccagct ccgggcggtc gcgcctggtg
    12361 ccggtgatct tctcggaaaa cagcttggtg cagccggccg cgtgcagttc ggcccgttgg
    12421 ttggtcaagt cctggtcgtc ggtgctgacg cgggcatagc ccagcaggcc agcggcggcg
    12481 ctcttgttca tggcgtaatg tctccggttc tagtcgcaag tattctactt tatgcgacta
    12541 aaacacgcga caagaaaacg ccaggaaaag ggcagggcgg cagcctgtcg cgtaacttag
    12601 gacttgtgcg acatgtcgtt ttcagaagac ggctgcactg aacgtcagaa gccgactgca
    12661 ctatagcagc ggaggggttg gatcaaagta ctttaaagta ctttaaagta ctttaaagta
    12721 ctttgatccc gaggggaacc ctgtggttgg catgcacata caaatggacg aacggataaa
    12781 ccttttcacg cccttttaaa tatccgatta ttctaataaa cgctcttttc tcttaggttt
    12841 acccgccaat atatcctgtc aaacactgat agtttaaact gaaggcggga aacgacaatc
    12901 tgatccaagc tcaagctaag ctagg
//
