LOCUS       sequence-447649-        8451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCMV and pT7 Human expression plasmid for SpCas9 enzyme with KKKCMN 
            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(KKKCMN)-P2A-EGFP (AHK131)
FEATURES             Location/Qualifiers
     source          1..8451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(70..87)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(241..829)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(321..340)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1000..1791,1792..1860))
                     /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     1623..1642
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1861..1965)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2044..2063)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2235..2614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2615..2818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2769..2789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2860..2879
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2860..2878
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2886..2895
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2892..6995
                     /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
                     ESILPKRNSDKLIARKKDWDPKKYGGFKKPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAKCLQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKMYNSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7050..7070
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7083..7148
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7167..7223
                     /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(7224..7226,7227..7229,7230..7943)
                     /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(7224..7226,7227..7229,7230..7940)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7530..7549)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7877..7898
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7949..7966
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7989..8006)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7995..8219
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8290..8306)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8290..8306)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8303..8325)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8314..8330
                     /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(8338..8344,8345..8362,8363..8368))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8383..8404
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
       61 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
      121 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
      181 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
      241 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
      301 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      361 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      421 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      481 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      541 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      601 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      661 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      721 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      781 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      841 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      901 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      961 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     1021 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     1081 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     1141 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     1201 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1261 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     1321 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1381 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1441 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1501 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1561 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1621 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1681 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1741 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1801 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1861 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     1921 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     1981 agtgccacct gacgtcgacg gatcgggaga tcgatctccc gatcccctag ggtcgactct
     2041 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     2101 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     2161 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     2221 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     2281 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     2341 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     2401 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     2461 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     2521 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     2581 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     2641 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     2701 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     2761 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
     2821 tttagtgaac cgtcagatcc gctagagatc cgcggccgct aatacgactc actataggga
     2881 gagccgccac catggacaag aagtacagca tcggcctgga catcggcacc aactctgtgg
     2941 gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca
     3001 acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg
     3061 aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga
     3121 accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct
     3181 tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc
     3241 ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc
     3301 acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg
     3361 ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg
     3421 acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg
     3481 aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga
     3541 gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc
     3601 tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg
     3661 acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca
     3721 acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt
     3781 ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc
     3841 tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag
     3901 ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga
     3961 acggctacgc cggctacatt gacggcggag ccagccagga agagttctac aagttcatca
     4021 agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg
     4081 acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg
     4141 gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc
     4201 gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca
     4261 ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga
     4321 acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca
     4381 acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt
     4441 acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc
     4501 ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc
     4561 ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact
     4621 ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc
     4681 tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg
     4741 aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga
     4801 aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca
     4861 ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca
     4921 agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga
     4981 tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg
     5041 gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca
     5101 tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg
     5161 agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca
     5221 gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga
     5281 aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc
     5341 agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg
     5401 atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc
     5461 tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga
     5521 agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt
     5581 tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca
     5641 tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact
     5701 cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca
     5761 ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg
     5821 agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc
     5881 tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg
     5941 acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact
     6001 tcttctacag caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga
     6061 tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg
     6121 gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa
     6181 agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg
     6241 ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc ttcaagaagc
     6301 ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac
     6361 tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga
     6421 atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca
     6481 agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg
     6541 ccaagtgcct gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt
     6601 acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc
     6661 tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct
     6721 ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc
     6781 accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca
     6841 atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagatgtaca
     6901 acagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg
     6961 agacacggat cgacctgtct cagctgggag gcgacggatc cggcggaggc ggaagcggga
     7021 aaagaaccgc cgacggcagc gaattcgagc ccaagaagaa gaggaaagtc tcgagcggag
     7081 gcgactacaa agaccatgac ggtgattata aagatcatga catcgattac aaggatgacg
     7141 atgacaaggg aggcggtgga agcggcgcaa caaacttctc tctgctgaaa caagccggag
     7201 atgtcgaaga gaatcctgga ccgatggtga gcaagggcga ggagctgttc accggggtgg
     7261 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     7321 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     7381 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     7441 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     7501 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     7561 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     7621 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     7681 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     7741 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     7801 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     7861 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     7921 gcatggacga gctgtacaag tgaccggtca tcatcaccat caccattgag tttaaacccg
     7981 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     8041 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     8101 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     8161 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     8221 ttctgaggcg gaaagaacca gctggggctc gataccgtcg acctctagct agagcttggc
     8281 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     8341 catacgagcc ggaagcataa agtgtaaagc ctagggtgcc taatgagtga gctaactcac
     8401 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt g
//
