pCSIIbsr-6013x

LOCUS       pCSIIbsr-6013x               13660 bp ds-DNA     circular SYN 10-AUG-2018
DEFINITION  An HRas biosensor based on fluorescence resonance energy transfer 
            (FRET) and those based on bioluminescence energy transfer (BRET).
ACCESSION   .
VERSION     .
KEYWORDS    pCSIIbsr-6013x
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13660)
  AUTHORS   Komatsu N, Terai K, Imanishi A, Kamioka Y, Sumiyama K, Jin T, Okada 
            Y, Nagai T, Matsuda M
  TITLE     A platform of BRET-FRET hybrid biosensors for optogenetics, chemical
            screening, and in vivo imaging.
  JOURNAL   Sci Rep. 2018 Jun 12;8(1):8984. doi: 10.1038/s41598-018-27174-x.
  PUBMED    29895862
REFERENCE   2  (bases 1 to 13660)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 10, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..13660
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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."
     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"
     primer_bind     complement(5678..5699)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5939..5958)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    7326..7912
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7493..7510)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7720..7739
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7913..8311
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8355..8943
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8408..8428)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8826..8837)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9462..9642
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9668..9685)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9674..9898
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9944..10372
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10031..10050)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10241..10262
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10381..10401)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10386..10715
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10566..10701
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10628..10647
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10763..10810
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10829..11203
                     /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    11333..11454
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11370..11389)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11424..11443
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11503..11519)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11503..11519)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11516..11538)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11527..11543
                     /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(11551..11581)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11596..11617
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11734..11751)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11905..12493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11985..12004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(12664..13524)
                     /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     13287..13306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13525..13629)
                     /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 acctcgagat
     4501 gacggaatat aagctggtgg tggtgggcgc cggcggtgtg ggcaagagtg cgctgaccat
     4561 ccagctgatc cagaaccatt ttgtggacga atacgacccc actatagagg attcctaccg
     4621 gaagcaggtg gtcattgatg gggagacgtg cctgttggac atcctggata ccgccggcca
     4681 ggaggagtac agcgccatgc gggaccagta catgcgcacc ggggagggct tcctgtgtgt
     4741 gtttgccatc aacaacacca agtcttttga ggacatccac cagtacaggg agcagatcaa
     4801 acgggtgaag gactcggatg acgtgcccat ggtgctggtg gggaacaagt gtgacctggc
     4861 tgcacgcact gtggaatctc ggcaggctca ggacctcgcc cgaagctacg gcatccccta
     4921 catcgagacc tcggccaaga cccggcaggg agtggaggat gccttctaca cgttggtgcg
     4981 tgagatccgg cagcacaagc tgcggaagct gaacggctcc ggaggaacca gtgctggtgg
     5041 tagtgctggt ggtagtgctg gtggtagtgc tggtggtagt gctggtggtt ccggcagtgc
     5101 tggtggtagt gctggtggta gtaccagtgc tggtggtagt gctggtggta gtgctggtgg
     5161 tagtgctggt ggtagtgctg gtggttccgg cagtgctggt ggtagtgctg gtggtagtac
     5221 cagtgctggt ggtagtgctg gtggtagtgc tggtggtagt gctggtggta gtgctggtgg
     5281 ttccggcagt gctggtggta gtgctggtgg tagtaccagt gctggtggta gtgctggtgg
     5341 tagtgctggt ggtagtgctg gtggtagtgc tggtggtacc ccttctaaga caagcaacac
     5401 tatccgtgtt ttcttgccga acaagcaaag aacagtggtc aatgtgcgaa atggaatgag
     5461 cttgcatgac tgccttatga aagcactcaa ggtgaggggc ctgcaaccag agagctgtgc
     5521 agtgttcaga cttctccacg aacacaaagg taaaaaagca cgcttagatt ggaatactga
     5581 agctgcgtct ttgattggag aagaacttca cgtagatttc ctgggcggcc gcatggtgag
     5641 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     5701 caacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     5761 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     5821 caccctgtcc tggggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     5881 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     5941 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     6001 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     6061 gtacaactac ttcagcggga acgtctatat caccgccgac aagcagaaga acggcatcaa
     6121 ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta
     6181 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     6241 cacccagtcc gccttaagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     6301 gttcttgacc gccgccgggg gtaccaaggt gtacgacccc gagcagagga agaggatgat
     6361 caccggcccc cagtggtggg ccaggtgcaa gcagatgaac gtgctggaca gcttcatcaa
     6421 ctactacgac agcgagaagc acgccgagaa cgccgtgatc ttcctgcacg gcaacgccac
     6481 tagcagctac ctgtggaggc acgtggtgcc ccacatcgag cccgtggcca ggtgcatcat
     6541 ccccgatctg atcggcatgg gcaagagcgg caagagcggc aacggcagct acaggctgct
     6601 ggaccactac aagtacctga ccgcctggtt cgagcttctg aacctgccca agaagatcat
     6661 cttcgtgggc cacgactggg gcgccgccct ggccttccac tacgcctacg agcaccagga
     6721 caggatcaag gccatcgtgc acatggagag cgtggtggac gtgatcgaga gctgggacga
     6781 gtggccagac atcgaggagg acatcgccct gatcaagagc gaggagggcg agaagatggt
     6841 gctggagaac aacttcttcg tggagaccgt gctgcccagc aagatcatga gaaagctgga
     6901 gcccgaggag ttcgccgcct acctggagcc cttcaaggag aagggcgagg tgagaagacc
     6961 caccctgagc tggcccagag agatccccct ggtgaagggc ggcaagcccg acgtggtgca
     7021 gatcgtgaga aactacaacg cctacctgag agccagcgac gacctgccca agctgttcat
     7081 cgagggcgac cccggcttct tcagcaacgc catcgtggag ggcgccaaga agttccccaa
     7141 caccgagttc gtgaaggtga agggcctgca cttcctccag gaggacgccc ccgacgagat
     7201 gggcaagtac atcaagagct tcgtggagag agtgctgaag aacgagcagt ctagaaagat
     7261 gagcaaagat ggtaaaaaga agaaaaagaa gtcaaagaca aagtgtgtaa ttatgtaagg
     7321 atccgcccct ctccctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     7381 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     7441 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     7501 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     7561 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     7621 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     7681 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     7741 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     7801 cggtacacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     7861 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggccaa
     7921 gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat
     7981 ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt
     8041 cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg
     8101 cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa
     8161 caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg
     8221 gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt
     8281 gctgccctct ggttatgtgt gggagggcta agtcgagcct ctagagttaa catcgaggga
     8341 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     8401 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     8461 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     8521 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     8581 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     8641 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     8701 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     8761 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     8821 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     8881 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     8941 cgcatcgata ccgtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag
     9001 ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag
     9061 tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     9121 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatatcc
     9181 ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaac tacacaccag
     9241 ggccagggat cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc
     9301 aagagaaggt agaagaagcc aatgaaggag agaacacccg cttgttacac cctgtgagcc
     9361 tgcatgggat ggatgacccg gagagagaag tattagagtg gaggtttgac agccgcctag
     9421 catttcatca catggcccga gagctgcatc cggactgtac tgggtctctc tggttagacc
     9481 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     9541 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     9601 gatccctcag acccttttag tcagtgtgga aaatctctag cagggcccgt ttaaacccgc
     9661 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     9721 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     9781 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     9841 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     9901 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     9961 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
    10021 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
    10081 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
    10141 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
    10201 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
    10261 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
    10321 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
    10381 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
    10441 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
    10501 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
    10561 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
    10621 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
    10681 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
    10741 ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt
    10801 ataatacgac aaggtgagga actaaaccat ggccaagttg accagtgccg ttccggtgct
    10861 caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg
    10921 ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag
    10981 cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct
    11041 ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg
    11101 gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc
    11161 ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt
    11221 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
    11281 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
    11341 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
    11401 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
    11461 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
    11521 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
    11581 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
    11641 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
    11701 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
    11761 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
    11821 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
    11881 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
    11941 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
    12001 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
    12061 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
    12121 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
    12181 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
    12241 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
    12301 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
    12361 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
    12421 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
    12481 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
    12541 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
    12601 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
    12661 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
    12721 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
    12781 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
    12841 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
    12901 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
    12961 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
    13021 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
    13081 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
    13141 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
    13201 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
    13261 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
    13321 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
    13381 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
    13441 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
    13501 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
    13561 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
    13621 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//