LOCUS       sequence-362675-        8451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV and T7 promoter expression plasmid for human codon usage 
            nSpRY(D10A) with a C-terminal bi-partite NLS, 3x FLAG, and P2A-eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8451)
  AUTHORS   Alonso-Lerma B, Jabalera Y, Samperio S, Morin M, Fernandez A, Hille 
            LT, Silverstein RA, Quesada-Ganuza A, Reifs A, Fernandez-Penalver S,
            Benitez Y, Soletto L, Gavira JA, Diaz A, Vranken W, Sanchez-Mejias 
            A, Guell M, Mojica FJM, Kleinstiver BP, Moreno-Pelayo MA, Montoliu 
            L, Perez-Jimenez R
  TITLE     Evolution of CRISPR-associated endonucleases as inferred from 
            resurrected proteins.
  JOURNAL   Nat Microbiol. 2023 Jan;8(1):77-90. doi: 10.1038/s41564-022-01265-y.
            Epub 2023 Jan 2.
  PUBMED    36593295
REFERENCE   2  (bases 1 to 8451)
  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/s41564-022-01265-y"; journalName: "Nat Microbiol"; date: 
            "2023-01"; volume: "8"; issue: "1"; pages: "77-90"
COMMENT     Sequence Label: pCMV-T7-nSpRY(D10A)-BPNLS(SV40)-3xFLAG-P2A-EGF...
FEATURES             Location/Qualifiers
     source          1..8451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        137..340
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     291..311
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     382..401
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        382..400
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      408..417
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             414..4517
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAERTRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESIRPKRNSDKLIARKKDWDPKKYGGFLWPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAKQLQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTRLGAPRAFKYFDTTIDPKQYRSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4572..4592
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4605..4670
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4689..4745
                     /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             join(4746..4748,4749..4751,4752..5465)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4746..4748,4749..4751,4752..5462)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5052..5071)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5399..5420
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5471..5488
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5511..5528)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5517..5741
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5812..5828)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5812..5828)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5825..5847)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5836..5852
                     /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(5860..5866,5867..5884,5885..5890))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5905..5926
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6043..6060)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6214..6802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6294..6313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6973..7764,7765..7833))
                     /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     7596..7615
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7834..7938)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8017..8036)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(8208..8451,1..136)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
       61 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      121 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      181 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      241 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      301 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      361 ccgctagaga tccgcggccg ctaatacgac tcactatagg gagagccgcc accatggaca
      421 agaagtacag catcggcctg gccatcggca ccaactctgt gggctgggcc gtgatcaccg
      481 acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca
      541 tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gagagaaccc
      601 ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc
      661 aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag
      721 agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg
      781 tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg
      841 tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca
      901 agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca
      961 agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg
     1021 ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg
     1081 aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg
     1141 ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca
     1201 aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg
     1261 gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga
     1321 gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca
     1381 agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc
     1441 tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca
     1501 ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga
     1561 tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc
     1621 ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg cacgccattc
     1681 tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga
     1741 tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg
     1801 cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg
     1861 acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc
     1921 ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg
     1981 agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg
     2041 agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc
     2101 agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg
     2161 tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg
     2221 acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc
     2281 tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt
     2341 tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga
     2401 gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc
     2461 tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga
     2521 cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc
     2581 acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg
     2641 tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa
     2701 tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc
     2761 ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa
     2821 acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt
     2881 acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc
     2941 ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga
     3001 accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact
     3061 ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg
     3121 ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg
     3181 aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt
     3241 acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg
     3301 tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc
     3361 acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta
     3421 agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg
     3481 ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca
     3541 tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga
     3601 tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg
     3661 tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag
     3721 gcggcttcag caaagagtct atcagaccca agaggaacag cgataagctg atcgccagaa
     3781 agaaggactg ggaccctaag aagtacggcg gcttcctgtg gcccaccgtg gcctattctg
     3841 tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc
     3901 tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg
     3961 aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc
     4021 tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccaagcag ctgcagaagg
     4081 gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg
     4141 agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca
     4201 agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg
     4261 ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca
     4321 gagagcaggc cgagaatatc atccacctgt ttaccctgac cagactggga gcccctagag
     4381 ccttcaagta ctttgacacc accatcgacc ccaagcagta cagaagcacc aaagaggtgc
     4441 tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt
     4501 ctcagctggg aggcgacgga tccggcggag gcggaagcgg gaaaagaacc gccgacggca
     4561 gcgaattcga gcccaagaag aagaggaaag tctcgagcgg aggcgactac aaagaccatg
     4621 acggtgatta taaagatcat gacatcgatt acaaggatga cgatgacaag ggaggcggtg
     4681 gaagcggcgc aacaaacttc tctctgctga aacaagccgg agatgtcgaa gagaatcctg
     4741 gaccgatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     4801 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     4861 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     4921 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     4981 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     5041 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     5101 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     5161 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     5221 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     5281 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     5341 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     5401 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     5461 agtgaccggt catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt
     5521 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     5581 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     5641 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     5701 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     5761 cagctggggc tcgataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5821 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5881 aaagtgtaaa gcctagggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5941 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6001 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6061 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6121 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6181 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6241 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6301 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6361 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6421 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6481 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6541 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6601 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6661 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6721 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6781 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6841 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6901 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6961 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7021 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7081 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7141 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7201 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7261 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7321 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7381 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7441 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7501 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7561 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7621 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7681 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7741 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7801 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7861 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7921 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     7981 agatcgatct cccgatcccc tagggtcgac tctcagtaca atctgctctg atgccgcata
     8041 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     8101 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     8161 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
     8221 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     8281 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     8341 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     8401 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag t
//
