LOCUS       sequence-420955-       12247 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A FRET biosensor for ROCK activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12247)
  AUTHORS   Deguchi E, Lin S, Hirayama D, Matsuda K, Tanave A, Sumiyama K, 
            Tsukiji S, Otani T, Furuse M, Sorkin A, Matsuda M, Terai K
  TITLE     Low-affinity ligands of the epidermal growth factor receptor are 
            long-range signal transmitters in collective cell migration of 
            epithelial cells.
  JOURNAL   Cell Rep. 2024 Nov 14;43(11):114986. doi: 
            10.1016/j.celrep.2024.114986.
  PUBMED    39546398
REFERENCE   2  (bases 1 to 12247)
  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: "Cell Rep.";
            date: "2024-11-14"; volume: "43(11):114986. doi"; pages: " 
            10.1016/j.celrep.2024.114986"
COMMENT     Sequence Label: pCSIIbsr-Eevee-ROCK-NES
FEATURES             Location/Qualifiers
     source          1..12247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        616..814
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             832..1012
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1059..1184
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1676..1909
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2094..2138
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2412..2529
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2585..3747
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2816..3738
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3695..3715
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3782..4492
                     /codon_start=1
                     /product="yellow fluorescent protein optimized for FRET 
                     with CyPet (Nguyen and Daugherty, 2005)"
                     /label=YPet
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LLCTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLLE
                     FLTAAGITEGMNELY"
     CDS             join(5144..5146,5147..5149,5150..5662)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(5189..5210)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5450..5469)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5797..5818
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5873..5899
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     misc_feature    join(5913..6487,6488..6490,6491..6499)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6080..6097)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6307..6326
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6500..6898
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    6942..7530
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6995..7015)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7413..7424)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8049..8229
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8255..8272)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8261..8485
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8531..8959
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8618..8637)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8828..8849
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8968..8988)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8973..9302
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9153..9288
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9215..9234
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9350..9397
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9416..9790
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    9920..10041
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9957..9976)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10011..10030
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10090..10106)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10090..10106)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10103..10125)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10114..10130
                     /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)."
     promoter        complement(join(10138..10144,10145..10162,10163..10168))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10183..10204
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10321..10338)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10492..11080)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10572..10591)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11251..12042,12043..12111))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     11874..11893
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12112..12216)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc
      841 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      901 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      961 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     1021 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     1081 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     1141 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     1201 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     1261 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     1321 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     1381 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     1441 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     1501 gtaagaccac cgcacagcaa gggccgctga tcttcagacc tggaggagga gatatgaggg
     1561 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1621 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1681 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     1741 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1801 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1861 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1921 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1981 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     2041 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     2101 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     2161 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     2221 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     2281 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
     2341 gtggagagag agacagagac agatccattc gattagtgaa cggatctaca aatggcagta
     2401 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     2461 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     2521 caaaattttc gggtttatta cagggacagc agaaattcac tttgaattaa ttcaagcttc
     2581 gtgaggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     2641 ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga
     2701 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     2761 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag
     2821 tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg
     2881 aattacttcc acctggctcc agtacgtgat tcttgatccc gagctggagc caggggcggg
     2941 ccttgcgctt taggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg
     3001 ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct
     3061 ctagccattt aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt
     3121 gtaaatgcgg gccaggatct gcacactggt atttcggttt ttggggccgc gggcggcgac
     3181 ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg
     3241 agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg
     3301 ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg
     3361 gaaagatggc cgcttcccgg ccctgctcca gggggctcaa aatggaggac gcggcgctcg
     3421 ggagagcggg cgggtgagtc acccacacaa aggaaagggg cctttccgtc ctcagccgtc
     3481 gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctggagc
     3541 ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac
     3601 actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa
     3661 tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt
     3721 ttttttcttc catttcaggt gtcgtgaaca cgctaccggt ctcgacgaat tcgcccttag
     3781 catgtctaaa ggtgaagaat tattcactgg tgttgtccca attttggttg aattagatgg
     3841 tgatgttaat ggtcacaaat tttctgtctc cggtgaaggt gaaggtgatg ctacgtacgg
     3901 taaattgacc ttaaaattac tctgtactac tggtaaattg ccagttccat ggccaacctt
     3961 agtcactact ttaggttatg gtgttcaatg ttttgctaga tacccagatc atatgaaaca
     4021 acatgacttt ttcaagtctg ccatgccaga aggttatgtt caagaaagaa ctattttttt
     4081 caaagatgac ggtaactaca agaccagagc tgaagtcaag tttgaaggtg ataccttagt
     4141 taatagaatc gaattaaaag gtattgattt taaagaagat ggtaacattt taggtcacaa
     4201 attggaatac aactataact ctcacaatgt ttacatcact gctgacaaac aaaagaatgg
     4261 tatcaaagct aacttcaaaa ttagacacaa cattgaagat ggtggtgttc aattagctga
     4321 ccattatcaa caaaatactc caattggtga tggtccagtc ttgttaccag acaaccatta
     4381 cttatcctat caatctgcct tattcaaaga tccaaacgaa aagagagacc acatggtctt
     4441 gttagaattt ttgactgctg ctggtattac cgagggtatg aatgaattgt acctcgagaa
     4501 gttttctcaa gaacagatcg gcgaaaacat tgtgtgcagg gtcatttgta ccacgggtca
     4561 aattcccatc cgagatttgt cagctgatat ttcacaagtg cttaaggaaa aacgatccat
     4621 aaagaaagtt tggacatttg gtagaaaccc agcctgtgac tatcatttag gaaacatttc
     4681 aagactgtca aataagcatt tccaaatact actaggagaa gacggtaacc ttttattgaa
     4741 tgacatttcc actaatggga cctggttaaa tgggcaaaaa gtcgagaaga acagcaatca
     4801 gttactgtcc caaggtgatg aaataaccgt tggtgtaggc gtggaatcag atattttatc
     4861 tctggtcatt ttcataaacg acaaatttaa gcagtgcctg gagcagaaca aagttgatcg
     4921 cggtaccagt gctggtggta gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc
     4981 tggtggttcc ggcagtgctg gtggtagtgc tggtggtagt accagtgctg gtggtagtgc
     5041 tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggttccggaa agagaagaaa
     5101 cagaagaaag accctggtgg acctgcctct ggatggcggc cgcatggtga gcaagggcga
     5161 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     5221 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     5281 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     5341 ctggggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     5401 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     5461 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     5521 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     5581 catcagccac aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt
     5641 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     5701 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacttga gcacccagtc
     5761 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     5821 cgccgccggg atcactctcg gcatggacga gctgggccgc tctagactgc agctgcctcc
     5881 tctggaacgc ctgactctgg attaaggatc cgcccctctc cctccccccc ccctaacgtt
     5941 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     6001 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     6061 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     6121 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     6181 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     6241 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     6301 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     6361 cattgtatgg gatctgatct ggggcctcgg tacacatgct ttacatgtgt ttagtcgagg
     6421 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     6481 tgataatatg gccacaacca tggccaagcc tttgtctcaa gaagaatcca ccctcattga
     6541 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc
     6601 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg
     6661 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac
     6721 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg
     6781 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca
     6841 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagt
     6901 cgagcctcta gagttaacat cgagggatca agcttatcga taatcaacct ctggattaca
     6961 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     7021 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     7081 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     7141 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     7201 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     7261 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     7321 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     7381 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     7441 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     7501 gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgagaccta gaaaaacatg
     7561 gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac
     7621 aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt
     7681 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     7741 ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact
     7801 tccctgattg gcagaactac acaccagggc cagggatcag atatccactg acctttggat
     7861 ggtgctacaa gctagtacca gttgagcaag agaaggtaga agaagccaat gaaggagaga
     7921 acacccgctt gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat
     7981 tagagtggag gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg
     8041 actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     8101 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     8161 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     8221 tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     8281 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     8341 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     8401 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     8461 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     8521 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     8581 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     8641 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     8701 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     8761 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     8821 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     8881 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     8941 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     9001 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     9061 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     9121 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     9181 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     9241 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     9301 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
     9361 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc
     9421 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt
     9481 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
     9541 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac
     9601 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
     9661 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
     9721 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
     9781 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     9841 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     9901 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     9961 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
    10021 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
    10081 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
    10141 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
    10201 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
    10261 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
    10321 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
    10381 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
    10441 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
    10501 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
    10561 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
    10621 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
    10681 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
    10741 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
    10801 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
    10861 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
    10921 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
    10981 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
    11041 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
    11101 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
    11161 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
    11221 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
    11281 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
    11341 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
    11401 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
    11461 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
    11521 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
    11581 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
    11641 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
    11701 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
    11761 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
    11821 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
    11881 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
    11941 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
    12001 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
    12061 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
    12121 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
    12181 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
    12241 tgacgtc
//
