LOCUS       sequence-447690-        8451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCMV and pT7 Human expression plasmid for SpCas9 enzyme with SRSCMV 
            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(SRSCMV)-P2A-EGFP (RTW3732)
FEATURES             Location/Qualifiers
     source          1..8451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(78..95)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(249..837)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(329..348)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1008..1799,1800..1868))
                     /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     1631..1650
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1869..1973)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2052..2071)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2243..2622
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2623..2826
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2777..2797
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2868..2887
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2868..2886
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2894..2903
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2900..7003
                     /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
                     ESILPKRNSDKLIARKKDWDPKKYGGFSRPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASASCLQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKMYVSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7058..7078
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7091..7156
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7175..7231
                     /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(7232..7234,7235..7237,7238..7951)
                     /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(7232..7234,7235..7237,7238..7948)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7538..7557)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7885..7906
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7957..7974
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7997..8014)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8003..8227
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8298..8314)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8298..8314)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8311..8333)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8322..8338
                     /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(8346..8352,8353..8370,8371..8376))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8391..8412
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
       61 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
      121 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
      181 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
      241 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
      301 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
      361 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
      421 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
      481 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
      541 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
      601 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
      661 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
      721 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      781 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
      841 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
      901 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
      961 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     1021 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     1081 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     1141 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     1201 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     1261 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     1321 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     1381 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     1441 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     1501 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     1561 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1621 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     1681 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1741 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1801 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1861 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1921 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     1981 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc gatctcccga tcccctaggg
     2041 tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     2101 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     2161 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
     2221 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
     2281 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     2341 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     2401 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     2461 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     2521 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     2581 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     2641 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     2701 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     2761 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     2821 agagctggtt tagtgaaccg tcagatccgc tagagatccg cggccgctaa tacgactcac
     2881 tatagggaga gccgccacca tggacaagaa gtacagcatc ggcctggaca tcggcaccaa
     2941 ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt
     3001 gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga
     3061 cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag
     3121 acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga
     3181 cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga
     3241 gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac
     3301 catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat
     3361 ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct
     3421 gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca
     3481 gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc
     3541 cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa
     3601 gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag
     3661 caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga
     3721 cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa
     3781 gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa
     3841 ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct
     3901 gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca
     3961 gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa
     4021 gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa
     4081 cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat
     4141 ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa
     4201 ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc
     4261 tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac
     4321 cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg
     4381 gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct
     4441 gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat
     4501 gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa
     4561 gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg
     4621 cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata
     4681 ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga
     4741 cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga
     4801 acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg
     4861 gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca
     4921 gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat
     4981 gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc
     5041 cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa
     5101 gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca
     5161 caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca
     5221 gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca
     5281 gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta
     5341 ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc
     5401 cgactacgat gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgacaa
     5461 caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga
     5521 ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca
     5581 gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc
     5641 cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat
     5701 cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa
     5761 agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa
     5821 agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg
     5881 aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa
     5941 ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc
     6001 caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa
     6061 cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg
     6121 ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat
     6181 cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag
     6241 gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt
     6301 cagcagaccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc
     6361 caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt
     6421 cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct
     6481 gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct
     6541 ggcctctgcc agctgcctgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa
     6601 cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca
     6661 gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag
     6721 cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta
     6781 caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac
     6841 cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa
     6901 gatgtacgtg agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg
     6961 cctgtacgag acacggatcg acctgtctca gctgggaggc gacggatccg gcggaggcgg
     7021 aagcgggaaa agaaccgccg acggcagcga attcgagccc aagaagaaga ggaaagtctc
     7081 gagcggaggc gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa
     7141 ggatgacgat gacaagggag gcggtggaag cggcgcaaca aacttctctc tgctgaaaca
     7201 agccggagat gtcgaagaga atcctggacc gatggtgagc aagggcgagg agctgttcac
     7261 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     7321 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     7381 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     7441 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     7501 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     7561 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     7621 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     7681 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     7741 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     7801 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     7861 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     7921 cactctcggc atggacgagc tgtacaagtg accggtcatc atcaccatca ccattgagtt
     7981 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     8041 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     8101 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     8161 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     8221 tctatggctt ctgaggcgga aagaaccagc tggggctcga taccgtcgac ctctagctag
     8281 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     8341 ccacacaaca tacgagccgg aagcataaag tgtaaagcct agggtgccta atgagtgagc
     8401 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa c
//
