LOCUS       sequence-447668-        8451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCMV and pT7 Human expression plasmid for SpCas9 enzyme with MRKCLN 
            amino acids at positions 1135,1136,1218,1219,1335,T1337 with C-term 
            BPNLS-P2A-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8451)
  AUTHORS   Silverstein RA, Kim N, Kroell AS, Walton RT, Delano J, Butcher RM, 
            Pacesa M, Smith BK, Christie KA, Ha LL, Meis RJ, Clark AB, Spinner 
            AD, Lazzarotto CR, Li Y, Matsubara A, Urbina EO, Dahl GA, Correia 
            BE, Marks DS, Tsai SQ, Pinello L, De Ravin SS, Liu Q, Kleinstiver BP
  TITLE     Custom CRISPR-Cas9 PAM variants via scalable engineering and machine
            learning.
  JOURNAL   Nature. 2025 Apr 22. doi: 10.1038/s41586-025-09021-y.
  PUBMED    40262634
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"; journalName: "Nature. 
            2025 Apr 22. doi: 10.1038/s41586-025-09021-y."
COMMENT     Sequence Label: pCMV-T7-SpCas9(MRKCLN)-P2A-EGFP (AHK130)
FEATURES             Location/Qualifiers
     source          1..8451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(56..73)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(227..815)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(307..326)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(986..1777,1778..1846))
                     /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     1609..1628
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1847..1951)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2030..2049)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2221..2600
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2601..2804
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2755..2775
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2846..2865
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2846..2864
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2872..2881
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2878..6981
                     /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
                     ESILPKRNSDKLIARKKDWDPKKYGGFMRPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAKCLQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKLYNSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7036..7056
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7069..7134
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7153..7209
                     /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(7210..7212,7213..7215,7216..7929)
                     /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(7210..7212,7213..7215,7216..7926)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7516..7535)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7863..7884
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7935..7952
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7975..7992)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7981..8205
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8276..8292)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8276..8292)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8289..8311)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8300..8316
                     /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(8324..8330,8331..8348,8349..8354))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8369..8390
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
       61 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
      121 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
      181 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
      241 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
      301 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
      361 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
      421 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
      481 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
      541 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
      601 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
      661 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
      721 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
      781 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
      841 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
      901 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
      961 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     1021 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     1081 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     1141 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     1201 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     1261 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     1321 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     1381 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     1441 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     1501 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     1561 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     1621 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     1681 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     1741 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     1801 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     1861 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     1921 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     1981 tcgacggatc gggagatcga tctcccgatc ccctagggtc gactctcagt acaatctgct
     2041 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     2101 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     2161 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
     2221 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     2281 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     2341 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     2401 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     2461 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     2521 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     2581 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     2641 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     2701 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     2761 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     2821 agatccgcta gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg
     2881 gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc
     2941 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
     3001 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
     3061 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
     3121 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
     3181 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
     3241 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
     3301 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
     3361 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
     3421 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
     3481 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
     3541 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg
     3601 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
     3661 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
     3721 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
     3781 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
     3841 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
     3901 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
     3961 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
     4021 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
     4081 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
     4141 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
     4201 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
     4261 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
     4321 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
     4381 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
     4441 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
     4501 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
     4561 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
     4621 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
     4681 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
     4741 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
     4801 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
     4861 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
     4921 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
     4981 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
     5041 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
     5101 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
     5161 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
     5221 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
     5281 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
     5341 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc
     5401 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac
     5461 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
     5521 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
     5581 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
     5641 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
     5701 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
     5761 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
     5821 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
     5881 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
     5941 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
     6001 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
     6061 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc
     6121 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
     6181 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc
     6241 agaaagaagg actgggaccc taagaagtac ggcggcttca tgagacccac cgtggcctat
     6301 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
     6361 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
     6421 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
     6481 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccaa gtgcctgcag
     6541 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
     6601 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
     6661 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
     6721 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc
     6781 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
     6841 gccgccttca agtactttga caccaccatc gaccggaagc tgtacaacag caccaaagag
     6901 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
     6961 ctgtctcagc tgggaggcga cggatccggc ggaggcggaa gcgggaaaag aaccgccgac
     7021 ggcagcgaat tcgagcccaa gaagaagagg aaagtctcga gcggaggcga ctacaaagac
     7081 catgacggtg attataaaga tcatgacatc gattacaagg atgacgatga caagggaggc
     7141 ggtggaagcg gcgcaacaaa cttctctctg ctgaaacaag ccggagatgt cgaagagaat
     7201 cctggaccga tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     7261 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     7321 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     7381 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     7441 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     7501 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     7561 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     7621 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     7681 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     7741 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     7801 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     7861 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     7921 tacaagtgac cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga
     7981 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     8041 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     8101 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     8161 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     8221 gaaccagctg gggctcgata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     8281 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     8341 gcataaagtg taaagcctag ggtgcctaat gagtgagcta actcacatta attgcgttgc
     8401 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa t
//
