LOCUS       sequence-447673-        8451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCMV and pT7 Human expression plasmid for SpCas9 enzyme with MRRWMR 
            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(MRRWMR)-P2A-EGFP (RAS2816)
FEATURES             Location/Qualifiers
     source          1..8451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(55..72)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(226..814)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(306..325)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(985..1776,1777..1845))
                     /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     1608..1627
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1846..1950)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2029..2048)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2220..2599
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2600..2803
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2754..2774
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2845..2864
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2845..2863
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2871..2880
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2877..6980
                     /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
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARWLQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKMYRSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7035..7055
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7068..7133
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7152..7208
                     /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(7209..7211,7212..7214,7215..7928)
                     /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(7209..7211,7212..7214,7215..7925)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7515..7534)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7862..7883
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7934..7951
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7974..7991)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7980..8204
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8275..8291)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8275..8291)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8288..8310)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8299..8315
                     /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(8323..8329,8330..8347,8348..8353))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8368..8389
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
       61 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
      121 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
      181 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      241 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      301 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      361 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      421 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      481 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      541 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
      601 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
      661 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
      721 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
      781 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
      841 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
      901 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
      961 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     1021 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     1081 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     1141 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     1201 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     1261 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     1321 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     1381 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     1441 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     1501 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     1561 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     1621 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     1681 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     1741 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     1801 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     1861 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     1921 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     1981 cgacggatcg ggagatcgat ctcccgatcc cctagggtcg actctcagta caatctgctc
     2041 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     2101 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     2161 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
     2221 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     2281 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     2341 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     2401 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     2461 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     2521 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     2581 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     2641 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     2701 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     2761 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
     2821 gatccgctag agatccgcgg ccgctaatac gactcactat agggagagcc gccaccatgg
     2881 acaagaagta cagcatcggc ctggacatcg gcaccaactc tgtgggctgg gccgtgatca
     2941 ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca
     3001 gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca
     3061 cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc
     3121 tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg
     3181 aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca
     3241 tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac
     3301 tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga
     3361 tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg
     3421 acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca
     3481 acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc
     3541 tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggaaacctga
     3601 ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg
     3661 ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga
     3721 tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc
     3781 tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga
     3841 tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc
     3901 agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct
     3961 acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa
     4021 agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc
     4081 agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca
     4141 ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga
     4201 agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat
     4261 tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg
     4321 tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc
     4381 tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata
     4441 acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg
     4501 gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga
     4561 agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg
     4621 gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca
     4681 aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga
     4741 ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc
     4801 tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc
     4861 tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt
     4921 tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc
     4981 tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg
     5041 agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga
     5101 aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg
     5161 aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga
     5221 agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg
     5281 aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata
     5341 tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccatatcg
     5401 tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca
     5461 agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact
     5521 actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca
     5581 aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg
     5641 tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta
     5701 agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc
     5761 tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc
     5821 accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc
     5881 ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga
     5941 tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca
     6001 tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc
     6061 tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca
     6121 ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga
     6181 caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca
     6241 gaaagaagga ctgggaccct aagaagtacg gcggcttcat gagacccacc gtggcctatt
     6301 ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag
     6361 agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc
     6421 tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact
     6481 ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccaga tggctgcaga
     6541 agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact
     6601 atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc
     6661 acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc
     6721 tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca
     6781 tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg
     6841 ccgccttcaa gtactttgac accaccatcg accggaagat gtacagaagc accaaagagg
     6901 tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc
     6961 tgtctcagct gggaggcgac ggatccggcg gaggcggaag cgggaaaaga accgccgacg
     7021 gcagcgaatt cgagcccaag aagaagagga aagtctcgag cggaggcgac tacaaagacc
     7081 atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac aagggaggcg
     7141 gtggaagcgg cgcaacaaac ttctctctgc tgaaacaagc cggagatgtc gaagagaatc
     7201 ctggaccgat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     7261 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     7321 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     7381 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     7441 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     7501 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     7561 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     7621 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     7681 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     7741 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     7801 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     7861 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     7921 acaagtgacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     7981 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     8041 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     8101 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     8161 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     8221 aaccagctgg ggctcgatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     8281 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     8341 cataaagtgt aaagcctagg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     8401 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat g
//
