LOCUS       sequence-447652-        8451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCMV and pT7 Human expression plasmid for SpCas9 enzyme with KWHMLM 
            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(KWHMLM)-P2A-EGFP (RAS2869)
FEATURES             Location/Qualifiers
     source          1..8451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(61..78)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(232..820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(312..331)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(991..1782,1783..1851))
                     /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     1614..1633
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1852..1956)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2035..2054)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2226..2605
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2606..2809
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2760..2780
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2851..2870
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2851..2869
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2877..2886
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2883..6986
                     /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
                     ESILPKRNSDKLIARKKDWDPKKYGGFKWPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAHMLQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKLYMSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7041..7061
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7074..7139
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7158..7214
                     /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(7215..7217,7218..7220,7221..7934)
                     /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(7215..7217,7218..7220,7221..7931)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7521..7540)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7868..7889
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7940..7957
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7980..7997)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7986..8210
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8281..8297)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8281..8297)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8294..8316)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8305..8321
                     /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(8329..8335,8336..8353,8354..8359))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8374..8395
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
       61 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
      121 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
      181 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
      241 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
      301 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
      361 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
      421 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
      481 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
      541 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
      601 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
      661 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
      721 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
      781 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
      841 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
      901 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
      961 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     1021 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     1081 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     1141 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     1201 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     1261 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     1321 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     1381 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     1441 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     1501 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     1561 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     1621 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     1681 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     1741 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     1801 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     1861 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     1921 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     1981 tgacgtcgac ggatcgggag atcgatctcc cgatccccta gggtcgactc tcagtacaat
     2041 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     2101 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     2161 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     2221 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     2281 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     2341 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2401 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2461 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     2521 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     2581 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     2641 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     2701 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     2761 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     2821 ccgtcagatc cgctagagat ccgcggccgc taatacgact cactataggg agagccgcca
     2881 ccatggacaa gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg
     2941 tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc
     3001 ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg
     3061 aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct
     3121 gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca
     3181 gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg
     3241 gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa
     3301 agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc
     3361 acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg
     3421 acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc
     3481 ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca
     3541 gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa
     3601 acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg
     3661 aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg
     3721 cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca
     3781 tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct
     3841 ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc
     3901 ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg
     3961 ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc
     4021 tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc
     4081 ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc
     4141 acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga
     4201 tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca
     4261 gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg
     4321 aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata
     4381 agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg
     4441 tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc
     4501 tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga
     4561 ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa
     4621 tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa
     4681 ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg
     4741 tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg
     4801 cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg
     4861 gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc
     4921 tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg
     4981 acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc
     5041 tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga
     5101 cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg
     5161 tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga
     5221 gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc
     5281 ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc
     5341 gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc
     5401 atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa
     5461 gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga
     5521 agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc
     5581 tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac
     5641 agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga
     5701 acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt
     5761 ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca
     5821 actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa
     5881 agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga
     5941 agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca
     6001 gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc
     6061 ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt
     6121 ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg
     6181 tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga
     6241 tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcaagtgg cccaccgtgg
     6301 cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg
     6361 tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg
     6421 actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta
     6481 agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct gcccacatgc
     6541 tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca
     6601 gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg
     6661 aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag
     6721 tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata
     6781 agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag
     6841 cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagctgtac atgagcacca
     6901 aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga
     6961 tcgacctgtc tcagctggga ggcgacggat ccggcggagg cggaagcggg aaaagaaccg
     7021 ccgacggcag cgaattcgag cccaagaaga agaggaaagt ctcgagcgga ggcgactaca
     7081 aagaccatga cggtgattat aaagatcatg acatcgatta caaggatgac gatgacaagg
     7141 gaggcggtgg aagcggcgca acaaacttct ctctgctgaa acaagccgga gatgtcgaag
     7201 agaatcctgg accgatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     7261 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     7321 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     7381 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     7441 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     7501 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     7561 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     7621 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     7681 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     7741 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     7801 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     7861 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     7921 agctgtacaa gtgaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc
     7981 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     8041 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     8101 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     8161 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     8221 ggaaagaacc agctggggct cgataccgtc gacctctagc tagagcttgg cgtaatcatg
     8281 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     8341 cggaagcata aagtgtaaag cctagggtgc ctaatgagtg agctaactca cattaattgc
     8401 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc a
//
