LOCUS       sequence-275524-        8451 bp    DNA     circular SYN 14-APR-2025
DEFINITION  CMV and T7 promoter expression plasmid for human codon optimized 
            SpCas9-VRQR(D1135V/G1218R/R1335Q/T1337R) with a c-terminal 
            bi-partite NLS, 3x flag tag, and P2A-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8451)
  AUTHORS   Walton RT, Christie KA, Whittaker MN, Kleinstiver BP
  TITLE     Unconstrained genome targeting with near-PAMless engineered 
            CRISPR-Cas9 variants.
  JOURNAL   Science. 2020 Apr 17;368(6488):290-296. doi: 
            10.1126/science.aba8853. Epub 2020 Mar 26.
  PUBMED    32217751
REFERENCE   2  (bases 1 to 8451)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1126/science.aba8853"; journalName: "Science"; date: 
            "2020-04-17- 17"; volume: "368"; issue: "6488"; pages: "290-296"
COMMENT     Sequence Label: pCMV-T7-SpCas9-VRQR-P2A-EGFP (RTW3161)
FEATURES             Location/Qualifiers
     source          1..8451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        186..565
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        566..769
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     720..740
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     811..830
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        811..829
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      837..846
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             843..4946
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5001..5021
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5034..5099
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5118..5174
                     /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(5175..5177,5178..5180,5181..5894)
                     /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(5175..5177,5178..5180,5181..5891)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5481..5500)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5828..5849
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5900..5917
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5940..5957)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5946..6170
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6241..6257)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6241..6257)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6254..6276)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6265..6281
                     /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(6289..6295,6296..6313,6314..6319))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6334..6355
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6472..6489)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6643..7231)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6723..6742)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7402..8193,8194..8262))
                     /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     8025..8044
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8263..8367)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(8446..8451,1..14))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
       61 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
      121 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
      181 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      241 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      301 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      361 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      421 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      481 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      541 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      601 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      661 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      721 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
      781 ccgtcagatc cgctagagat ccgcggccgc taatacgact cactataggg agagccgcca
      841 ccatggacaa gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg
      901 tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc
      961 ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg
     1021 aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct
     1081 gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca
     1141 gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg
     1201 gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa
     1261 agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc
     1321 acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg
     1381 acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc
     1441 ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca
     1501 gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa
     1561 acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg
     1621 aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg
     1681 cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca
     1741 tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct
     1801 ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc
     1861 ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg
     1921 ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc
     1981 tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc
     2041 ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc
     2101 acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga
     2161 tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca
     2221 gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg
     2281 aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata
     2341 agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg
     2401 tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc
     2461 tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga
     2521 ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa
     2581 tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa
     2641 ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg
     2701 tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg
     2761 cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg
     2821 gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc
     2881 tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg
     2941 acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc
     3001 tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga
     3061 cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg
     3121 tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga
     3181 gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc
     3241 ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc
     3301 gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc
     3361 atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa
     3421 gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga
     3481 agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc
     3541 tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac
     3601 agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga
     3661 acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt
     3721 ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca
     3781 actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa
     3841 agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga
     3901 agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca
     3961 gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc
     4021 ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt
     4081 ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg
     4141 tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga
     4201 tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgtgagc cccaccgtgg
     4261 cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg
     4321 tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg
     4381 actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta
     4441 agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccagagaac
     4501 tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca
     4561 gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg
     4621 aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag
     4681 tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata
     4741 agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag
     4801 cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagcagtac agaagcacca
     4861 aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga
     4921 tcgacctgtc tcagctggga ggcgacggat ccggcggagg cggaagcggg aaaagaaccg
     4981 ccgacggcag cgaattcgag cccaagaaga agaggaaagt ctcgagcgga ggcgactaca
     5041 aagaccatga cggtgattat aaagatcatg acatcgatta caaggatgac gatgacaagg
     5101 gaggcggtgg aagcggcgca acaaacttct ctctgctgaa acaagccgga gatgtcgaag
     5161 agaatcctgg accgatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     5221 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     5281 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     5341 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     5401 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     5461 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     5521 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     5581 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     5641 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     5701 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     5761 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     5821 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     5881 agctgtacaa gtgaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     5941 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     6001 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     6061 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     6121 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     6181 ggaaagaacc agctggggct cgataccgtc gacctctagc tagagcttgg cgtaatcatg
     6241 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6301 cggaagcata aagtgtaaag cctagggtgc ctaatgagtg agctaactca cattaattgc
     6361 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6421 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6481 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6541 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6601 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6661 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6721 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6781 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6841 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6901 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6961 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7021 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7081 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7141 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7201 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7261 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7321 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7381 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7441 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7501 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7561 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7621 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7681 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7741 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7801 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7861 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7921 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7981 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8041 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8101 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8161 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8221 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8281 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8341 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     8401 cggatcggga gatcgatctc ccgatcccct agggtcgact ctcagtacaa t
//
