LOCUS       sequence-235017-        8961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR-Sirius plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8961)
  AUTHORS   Ma H, Tu LC, Naseri A, Chung YC, Grunwald D, Zhang S, Pederson T
  TITLE     CRISPR-Sirius: RNA scaffolds for signal amplification in genome 
            imaging.
  JOURNAL   Nat Methods. 2018 Nov;15(11):928-931. doi: 
            10.1038/s41592-018-0174-0. Epub 2018 Oct 30.
  PUBMED    30377374
REFERENCE   2  (bases 1 to 8961)
  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: 
            "10.1038/s41592-018-0174-0"; journalName: "Nat Methods"; date: 
            "2018-11"; volume: "15"; issue: "11"; pages: "928-931"
COMMENT     Sequence Label: pPUR-mU6-sgRNA-Sirius-8XPP7
FEATURES             Location/Qualifiers
     source          1..8961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     426..444
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             522..1118
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(522..541)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1018..1038
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1128..1184
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1200..1910
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGIYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             1911..1931
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     LTR             2027..2260
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2332..2453
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2369..2388)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2423..2442
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2493..2628
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2648..2667)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2649..2667)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2677..2694)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2677..2693)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2686..2708)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2835..3290
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2922..2941)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3132..3153
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3316..3420
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3421..3489,3490..4281)
                     /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     complement(3639..3658)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4452..5040
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4941..4960
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5194..5211
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5328..5349
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5364..5369,5370..5387,5388..5394)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5402..5418
                     /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     5407..5429
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5426..5442
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5426..5442
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5461..5481
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5463..5481
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5509..5735
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5736..5916
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5963..6088
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6581..6814
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6999..7043
                     /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"
     promoter        7344..7657
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     7576..7596
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     CDS             8005..8310
                     /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     8189..8208
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_feature    8792..8909
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        join(8958..8961,1..507)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
ORIGIN
        1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
       61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
      121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
      181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
      241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
      301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
      361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
      421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
      481 gaccgaatca ccgacctctc tccccagggg gatccgccac catgaccgag tacaagccca
      541 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
      601 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
      661 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
      721 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
      781 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
      841 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
      901 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
      961 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     1021 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     1081 gaccgcgcac ctggtgcatg acccgcaagc ccggtgccgc tagcggtgct actaacttca
     1141 gcctgctgaa gcaggctgga gacgtggagg agaaccctgg acctggtagt ggaacgcgta
     1201 tggtgtctaa gggcgaagag ctgattaagg agaacatgca catgaagctg tacatggagg
     1261 gcaccgtgga caaccatcac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg
     1321 gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc gccttcgaca
     1381 tcctggctac tagcttcctc tacggcagca agaccttcat caaccacacc cagggcatcc
     1441 ccgacttctt caagcagtcc ttccctgagg gcttcacatg ggagagagtc accacatacg
     1501 aagatggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct
     1561 acaacgtcaa gatcagaggg gtgaacttca catccaacgg ccctgtgatg cagaagaaaa
     1621 cactcggctg ggaggccttc accgagacgc tgtaccccgc tgacggcggc ctggaaggca
     1681 gaaacgacat ggccctgaag ctcgtgggcg ggagccatct gatcgcaaac gccaagacca
     1741 catatagatc caagaaaccc gctaagaacc tcaagatgcc tggcatctac tatgtggact
     1801 acagactgga aagaatcaag gaggccaaca acgagaccta cgtcgagcag cacgaggtgg
     1861 cagtggccag atactgcgac ctccctagca aactggggca caagcttaat ccaaagaaaa
     1921 agcggaaagt gtgagtcgac ctcgaggggg ggcccggtac ctttaagacc aatgacttac
     1981 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     2041 cactcccaac gaagaaaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
     2101 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     2161 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     2221 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt
     2281 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt
     2341 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2401 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     2461 tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2521 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     2581 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc
     2641 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg
     2701 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     2761 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     2821 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2881 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2941 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3001 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3061 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3121 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3181 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3241 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     3301 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3361 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3421 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     3481 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     3541 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     3601 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     3661 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3721 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3781 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     3841 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3901 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     3961 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     4021 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     4081 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4141 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4201 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4261 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     4321 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4381 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4441 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4501 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4561 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     4621 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4681 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4741 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4801 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4861 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4921 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4981 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5041 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5101 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     5161 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     5221 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     5281 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     5341 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     5401 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
     5461 gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg
     5521 caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag
     5581 agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag
     5641 gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag
     5701 atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct
     5761 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5821 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5881 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     5941 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     6001 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     6061 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     6121 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     6181 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     6241 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     6301 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     6361 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     6421 gcaagcggcc actgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     6481 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     6541 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     6601 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     6661 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     6721 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     6781 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     6841 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     6901 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     6961 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     7021 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     7081 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
     7141 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
     7201 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
     7261 gagacagatc cattcgatta gtgaacggat ctcgacggta tcgatcacga gactagcctc
     7321 gagcggccgc cctgtgctct agagatccga cgccgccatc tctaggcccg cgccggcccc
     7381 ctcgcacaga cttgtgggag aagctcggct actcccctgc cccggttaat ttgcatataa
     7441 tatttcctag taactataga ggcttaatgt gcgataaaag acagataatc tgttcttttt
     7501 aatactagct acattttaca tgataggctt ggatttctat aagagataca aatactaaat
     7561 tattatttta aaaaacagca caaaaggaaa ctcaccctaa ctgtaaagta attgtgtgtt
     7621 ttgagactat aaatatccct tggagaaaag ccttgtttgg tgtcttcgaa tgcatggctt
     7681 actaaaagcc agataacagt atgcgtattt gcgcgctgat ttttgcggta taagaatata
     7741 tactgatatg tatacccgaa gtatgtcaaa aagaggtgtg ctatgaagca gcgtattaca
     7801 gtgacagttg acagcgacag ctatcagttg ctcaaggcat atatgatgtc aatatctccg
     7861 gtctggtaag cacaaccatg cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag
     7921 cggaaaatca ggaagggatg gctgaggtcg cccggtttat tgaaatgaac ggctcttttg
     7981 ctgacgagaa cagggactgg tgaaatgcag tttaaggttt acacctataa aagagagagc
     8041 cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgcccgg gcgacggatg
     8101 gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg
     8161 gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg
     8221 gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac
     8281 gccattaacc tgatgttctg gggaatataa accctgcagg acgaagaccc gtttgagagc
     8341 tactgccatg aggagcagac gatatggcgt cgctccgatc cgaccagcag agcatatggg
     8401 ctcgctgggg ctgcagcagc agaggatatg gcctcgctgc ggagtgacga gcagaccata
     8461 tggggtcgct cggccaccag aagcagaaga tatggcttcg cttccggaca actagcagat
     8521 catatgggat cgctaggagt agagtagcag atgatatggc atcgctacgt gtgacaagca
     8581 gaacatatgg gttcgcttgt cacacaaact actcaaatgt ccgaaaggtg gcaaacactc
     8641 caaagcagcc aaacggatca aacatggcag tagcaagttc aaataaggct agtccgttat
     8701 caacttgaaa aagtggcacc gagtcggtgc tttttttatc gatggcaagg gaattctcga
     8761 cctcgagaca aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt
     8821 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
     8881 aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccac
     8941 tttggccgcg gctcgagggg g
//
