LOCUS       sequence-433014-        9416 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Encoding a Forster resonance energy transfer (FRET) biosensor for 
            Rac1 activity with cyan and yellow fluorescent protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9416)
  TITLE     Collective cell migration is coordinated by front-biased activation 
            of the feed-forward loop consisting of Ras, Rab5, and Rac1.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9416)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pPBbsr2-RaichuEV-Rac1HRasCT
FEATURES             Location/Qualifiers
     source          1..9416
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   674..708
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        1011..1390
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1392..1667
                     /label=chicken beta-actin promoter
     intron          1668..2684
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2608..2630
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2692..2711
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2748..3458
                     /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(4614..4616,4617..4619,4620..5132)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISGNVYITADKQKNGIK
                     ANFKIRHNIE"
     primer_bind     complement(4659..4680)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4920..4939)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    5480..6029
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5624..5641)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5851..5870
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(6500..6517)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6506..6730
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6777..6793)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6801..6817
                     /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(6825..6831,6832..6849,6850..6855))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6870..6891
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   7125..7187
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7228..7248)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7228..7246)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7267..7283)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7291..7307
                     /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(7315..7321,7322..7339,7340..7345))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7360..7381
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7498..7515)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7669..8257)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7749..7768)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8428..9219,9220..9288))
                     /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     9051..9070
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9289..9393)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggggcgcgc
      661 cattctagat taaccctaga aagatagtct gcgtaaaatt gacgcatgca ttcttgaaat
      721 attgctctct ctttctaaat agcgcgaatc cgtcgctgtg catttaggac atctcagtcg
      781 ccgcttggag ctcccgtgag gcgtgcttgt caatgcggta agtgtcactg attttgaact
      841 ataacgaccg cgtgagtcaa aatgacgcat gattatcttt tacgtgactt ttaagattta
      901 actcatacga taattatatt gttatttcat gttctactta cgtgataact tattatatat
      961 atattttctt gttatagata tcaactagaa tgctagcatg ggcccatctc gacattgatt
     1021 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     1081 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     1141 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     1201 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     1261 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     1321 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     1381 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
     1441 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
     1501 gggggggggg gggcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc
     1561 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
     1621 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc
     1681 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
     1741 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
     1801 agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc
     1861 tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg
     1921 tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg
     1981 cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc
     2041 ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt
     2101 gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag
     2161 ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg
     2221 ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg
     2281 ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg
     2341 cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt
     2401 tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc
     2461 gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt
     2521 cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct
     2581 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
     2641 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
     2701 tggttattgt gctgtctcat cattttggca aagaattcgc ccttagcatg tctaaaggtg
     2761 aagaattatt cactggtgtt gtcccaattt tggttgaatt agatggtgat gttaatggtc
     2821 acaaattttc tgtctccggt gaaggtgaag gtgatgctac gtacggtaaa ttgaccttaa
     2881 aattactctg tactactggt aaattgccag ttccatggcc aaccttagtc actactttag
     2941 gttatggtgt tcaatgtttt gctagatacc cagatcatat gaaacaacat gactttttca
     3001 agtctgccat gccagaaggt tatgttcaag aaagaactat ttttttcaaa gatgacggta
     3061 actacaagac cagagctgaa gtcaagtttg aaggtgatac cttagttaat agaatcgaat
     3121 taaaaggtat tgattttaaa gaagatggta acattttagg tcacaaattg gaatacaact
     3181 ataactctca caatgtttac atcactgctg acaaacaaaa gaatggtatc aaagctaact
     3241 tcaaaattag acacaacatt gaagatggtg gtgttcaatt agctgaccat tatcaacaaa
     3301 atactccaat tggtgatggt ccagtcttgt taccagacaa ccattactta tcctatcaat
     3361 ctgccttatt caaagatcca aacgaaaaga gagaccacat ggtcttgtta gaatttttga
     3421 ctgctgctgg tattaccgag ggtatgaatg aattgtacct cgagaaagag aaagagcggc
     3481 cagagatttc tctcccttca gattttgaac acacaattca tgtcggtttt gatgctgtca
     3541 caggggagtt tacgggaatg ccagagcagt gggcccgctt gcttcagaca tcaaatatca
     3601 ctaagtcgga gcagaagaaa aacccgcagg ctgttctgga tgtgttggag ttttacaact
     3661 cgaagaagac atccaacagc cagaaataca tgagctttac agataagtca gcttccggag
     3721 gaaccagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggtagtgctg
     3781 gtggttccgg cagtgctggt ggtagtgctg gtggtagtac cagtgctggt ggtagtgctg
     3841 gtggtagtgc tggtggtagt gctggtggta gtgctggtgg ttccggcagt gctggtggta
     3901 gtgctggtgg tagtaccagt gctggtggta gtgctggtgg tagtgctggt ggtagtgctg
     3961 gtggtagtgc tggtggttcc ggcagtgctg gtggtagtgc tggtggtagt accagtgctg
     4021 gtggtagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggtaccatgc
     4081 aggccatcaa gtgtgtggtg gtgggagacg gagctgtagg taaaacttgc ctactgatca
     4141 gttacacaac caatgcattt cctggagaat atatccctac tgtctttgac aattattctg
     4201 ccaatgttat ggtagatgga aaaccggtga atctgggctt atgggataca gctggacaag
     4261 aagattatga cagattacgc cccctatcct atccgcaaac agatgtgttc ttaatttgct
     4321 tttcccttgt gagtcctgca tcatttgaaa atgtccgtgc aaagtggtat cctgaggtgc
     4381 ggcaccactg tcccaacact cccatcatcc tagtgggaac taaacttgat cttagggatg
     4441 ataaagacac gatcgagaaa ctgaaggaga agaagctgac tcccatcacc tatccgcagg
     4501 gtctagccat ggctaaggag attggtgctg taaaatacct ggagtgctcg gcgctcacac
     4561 agcgaggcct caagacagtg tttgacgaag cgatccgagc agtccgcggc cgcatggtga
     4621 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     4681 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     4741 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     4801 ccaccctgtc ctggggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
     4861 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     4921 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     4981 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     5041 agtacaacta catcagcggg aacgtctata tcaccgccga caagcagaag aacggcatca
     5101 aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact
     5161 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     5221 gcacccagtc cgccttaagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     5281 agttcttgac cgccgccggg atcactctcg gcatggacga gctgtctaga aagctgaacc
     5341 ctcctgatga gagtggcccc ggctgcatga gctgcaagtg tgtgctctcc tgagtcgacg
     5401 ggccgcggta acaattgtta actaacttaa gctagcaacg gtttccctct agcgggatca
     5461 attccgcccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     5521 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     5581 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     5641 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     5701 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     5761 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     5821 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     5881 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     5941 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac
     6001 gtggttttcc tttgaaaaac acgataatac catggtcatg aaaacattta acatttctca
     6061 acaagatcta gaattagtag aagtagcgac agagaagatt acaatgcttt atgaggataa
     6121 taaacatcat gtgggagcgg caattcgtac gaaaacagga gaaatcattt cggcagtaca
     6181 tattgaagcg tatataggac gagtaactgt ttgtgcagaa gccattgcga ttggtagtgc
     6241 agtttcgaat ggacaaaagg attttgacac gattgtagct gttagacacc cttattctga
     6301 cgaagtagat agaagtattc gagtggtaag tccttgtggt atgtgtaggg agttgatttc
     6361 agactatgca ccagattgtt ttgtgttaat agaaatgaat ggcaagttag tcaaaactac
     6421 gattgaagaa ctcattccac tcaaatatac ccgaaattaa aagtcgagat gcatcgatga
     6481 tctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     6541 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     6601 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     6661 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     6721 ggctctatgg cttctgaggc ggaaagaacc tgcagcccaa gcttggcgta atcatggtca
     6781 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     6841 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     6901 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatctt aaaagttttg
     6961 ttactttata gaagaaattt tgagtttttg ttttttttta ataaataaat aaacataaat
     7021 aaattgtttg ttgaatttat tattagtatg taagtgtaaa tataataaaa cttaatatct
     7081 attcaaatta ataaataaac ctcgatatac agaccgataa aacacatgcg tcaattttac
     7141 gcatgattat ctttaacgta cgtcacaata tgattatctt tctagggtta atctagtata
     7201 cgcgtttaat taatccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     7261 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     7321 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     7381 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     7441 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     7501 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7561 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7621 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7681 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7741 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7801 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7861 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7921 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7981 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     8041 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     8101 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     8161 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     8221 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     8281 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     8341 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     8401 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     8461 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     8521 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     8581 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     8641 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     8701 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     8761 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     8821 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8881 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8941 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     9001 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     9061 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     9121 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     9181 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     9241 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     9301 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9361 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccac
//
