LOCUS       sequence-210276-        8508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  D16A+H588A mutant of Cas9 from Nm8013 cloned into pET28b with a 
            C-terminal Flag tag for protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8508)
  AUTHORS   Rousseau BA, Hou Z, Gramelspacher MJ, Zhang Y
  TITLE     Programmable RNA Cleavage and Recognition by a Natural CRISPR-Cas9 
            System from Neisseria meningitidis.
  JOURNAL   Mol Cell. 2018 Mar 1;69(5):906-914.e4. doi: 
            10.1016/j.molcel.2018.01.025. Epub 2018 Feb 15.
  PUBMED    29456189
REFERENCE   2  (bases 1 to 8508)
  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"; doi: 
            "10.1016/j.molcel.2018.01.025"; journalName: "Mol Cell"; date: 
            "2018-03-1- 1"; volume: "69"; issue: "5"; pages: "906-914.e4"
COMMENT     Sequence Label: pET28b-NmCas9-Flag D16A+H588A
FEATURES             Location/Qualifiers
     source          1..8508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     99..118
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        99..117
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    118..142
                     /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)."
     RBS             157..179
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             186..3431
                     /codon_start=1
                     /product="Cas9 endonuclease from the Neisseria meningitidis
                     Type II CRISPR/Cas system (Esvelt et al., 2013)"
                     /label=NmCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MAAFKPNSINYILGLAIGIASVGWAMVEIDEEENPIRLIDLGVRV
                     FERAEVPKTGDSLAMARRLARSVRRLTRRRAHRLLRTRRLLKREGVLQAANFDENGLIK
                     SLPNTPWQLRAAALDRKLTPLEWSAVLLHLIKHRGYLSQRKNEGETADKELGALLKGVA
                     GNAHALQTGDFRTPAELALNKFEKESGHIRNQRSDYSHTFSRKDLQAELILLFEKQKEF
                     GNPHVSGGLKEGIETLLMTQRPALSGDAVQKMLGHCTFEPAEPKAAKNTYTAERFIWLT
                     KLNNLRILEQGSERPLTDTERATLMDEPYRKSKLTYAQARKLLGLEDTAFFKGLRYGKD
                     NAEASTLMEMKAYHAISRALEKEGLKDKKSPLNLSPELQDEIGTAFSLFKTDEDITGRL
                     KDRIQPEILEALLKHISFDKFVQISLKALRRIVPLMEQGKRYDEACAEIYGDHYGKKNT
                     EEKIYLPPIPADEIRNPVVLRALSQARKVINGVVRRYGSPARIHIETAREVGKSFKDRK
                     EIEKRQEENRKDREKAAAKFREYFPNFVGEPKSKDILKLRLYEQQHGKCLYSGKEINLG
                     RLNEKGYVEIDAALPFSRTWDDSFNNKVLVLGSENQNKGNQTPYEYFNGKDNSREWQEF
                     KARVETSRFPRSKKQRILLQKFDEDGFKERNLNDTRYVNRFLCQFVADRMRLTGKGKKR
                     VFASNGQITNLLRGFWGLRKVRAENDRHHALDAVVVACSTVAMQQKITRFVRYKEMNAF
                     DGKTIDKETGEVLHQKTHFPQPWEFFAQEVMIRVFGKPDGKPEFEEADTLEKLRTLLAE
                     KLSSRPEAVHEYVTPLFVSRAPNRKMSGQGHMETVKSAKRLDEGVSVLRVPLTQLKLKD
                     LEKMVNREREPKLYEALKARLEAHKDDPAKAFAEPFYKYDKAGNRTQQVKAVRVEQVQK
                     TGVWVRNHNGIADNATMVRVDVFEKGDKYYLVPIYSWQVAKGILPDRAVVQGKDEEDWQ
                     LIDDSFNFKFSLHPNDLVEVITKKARMFGYFASCHRGTGNINIRIHDLDHKIGKNGILE
                     GIGVKTALSFQKYQIDELGKEIRPCRLKKRPPVR"
     CDS             3432..3455
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3468..3488
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     primer_bind     complement(3541..3559)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3555..3602
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3639..4094
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3726..3745)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3936..3957
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4186..5001)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4186..5001)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4909..4928
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4993..5012
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5123..5711
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5612..5631
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5865..5882
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5897..6036
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5962..5981)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6122..6144
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6138..6329)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(7138..8220)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     8182..8201
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8221..8298)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(8504..8508,1..15)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
       61 gtccggcgta gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga
      121 attgtgagcg gataacaatt cccctctaga aataattttg tttaacttta agaaggagat
      181 ataccatggc tgccttcaaa cctaattcaa tcaactacat cctcggcctc gctatcggca
      241 tcgcatccgt cggctgggcg atggtagaaa ttgacgaaga agaaaacccc atccgcctga
      301 ttgatttggg cgtgcgcgta tttgagcgtg ccgaagtacc gaaaacaggc gactcccttg
      361 ccatggcaag gcgtttggcg cgcagtgttc gccgcctgac ccgccgtcgc gcccaccgcc
      421 tgcttcggac ccgccgccta ttgaaacgcg aaggcgtatt acaagccgcc aattttgacg
      481 aaaacggctt gattaaatcc ttaccgaata caccatggca acttcgcgca gccgcattag
      541 accgcaaact gacgccttta gagtggtcgg cagtcttgtt gcatttaatc aaacatcgcg
      601 gctatttatc gcaacggaaa aacgagggcg aaactgccga taaggagctt ggcgctttgc
      661 ttaaaggcgt agccggcaat gcccatgcct tacagacagg cgatttccgc acaccggccg
      721 aattggcttt aaataaattt gagaaagaaa gcggccatat ccgcaatcag cgcagcgatt
      781 attcgcatac gttcagccgc aaagatttac aggcggagct gattttgctg tttgaaaaac
      841 aaaaagaatt tggcaatccg catgtttcag gcggccttaa agaaggtatt gaaaccctac
      901 tgatgacgca acgccctgcc ctgtccggcg atgccgttca aaaaatgttg gggcattgca
      961 ccttcgaacc ggcagagccg aaagccgcta aaaacaccta cacagccgaa cgtttcatct
     1021 ggctgaccaa gctgaacaac ctgcgtattt tagagcaagg cagcgagcgg ccattgaccg
     1081 ataccgaacg cgccacgctt atggacgagc catacagaaa atccaaactg acttacgcac
     1141 aagcccgtaa gctgctgggt ttagaagata ccgccttttt caaaggcttg cgctatggta
     1201 aagacaatgc cgaagcctca acattgatgg aaatgaaggc ctaccatgcc atcagccgtg
     1261 cactggaaaa agaaggattg aaagacaaaa aatccccatt aaacctttct cccgaattac
     1321 aagacgaaat cggcacggca ttctccctgt tcaaaaccga tgaagacatt acaggccgtc
     1381 tgaaagaccg tatacagccc gaaatcttag aagcgctgtt gaaacacatc agcttcgata
     1441 agttcgtcca aatttccttg aaagcattgc gccgaattgt gcctctaatg gaacaaggca
     1501 aacgttacga tgaagcctgc gccgaaatct acggagacca ttacggcaag aagaatacgg
     1561 aagaaaagat ttatctgccg ccgattcccg ccgacgaaat ccgcaacccc gtcgtcttgc
     1621 gcgccttatc tcaagcacgt aaggtcatta acggcgtggt acgccgttac ggctccccag
     1681 ctcgtatcca tattgaaact gcaagggaag taggtaaatc gtttaaagac cgcaaagaaa
     1741 ttgagaaacg ccaagaagaa aaccgcaaag accgggaaaa agccgccgcc aaattccgag
     1801 agtatttccc caattttgtc ggagaaccca aatccaaaga tattctgaaa ctgcgcctgt
     1861 acgagcaaca acacggcaaa tgcctgtatt cgggcaaaga aatcaactta ggccgtctga
     1921 acgaaaaagg ctatgtcgaa atcgacgctg ccctgccgtt ctcgcgcaca tgggacgaca
     1981 gtttcaacaa taaagtactg gtattgggca gcgaaaacca aaacaaaggc aatcaaaccc
     2041 cttacgaata cttcaacggc aaagacaaca gccgcgaatg gcaggaattt aaagcgcgtg
     2101 tcgaaaccag ccgtttcccg cgcagtaaaa aacaacggat tctgctgcaa aaattcgatg
     2161 aagacggctt taaagaacgc aatctgaacg acacgcgcta cgtcaaccgt ttcctgtgtc
     2221 aatttgttgc cgaccgtatg cggctgacag gtaaaggcaa gaaacgtgtc tttgcatcca
     2281 acggacaaat taccaatctg ttgcgcggct tttggggatt gcgcaaagtg cgtgcggaaa
     2341 acgaccgcca tcacgccttg gacgccgtcg tcgttgcctg ctcgaccgtt gccatgcagc
     2401 agaaaattac ccgttttgta cgctataaag agatgaacgc gtttgacggt aaaaccatag
     2461 acaaagaaac aggagaagtg ctgcatcaaa aaacacactt cccacaacct tgggaatttt
     2521 tcgcacaaga agtcatgatt cgcgtcttcg gcaaaccgga cggcaaaccc gaattcgaag
     2581 aagccgatac cctagaaaaa ctgcgcacgt tgcttgccga aaaattatca tctcgccccg
     2641 aagccgtaca cgaatacgtt acgccactgt ttgtttcacg cgcgcccaat cggaagatga
     2701 gcgggcaagg gcatatggag accgtcaaat ccgccaaacg actggacgaa ggcgtcagcg
     2761 tgttgcgcgt accgctgaca cagttaaaac tgaaagactt ggaaaaaatg gtcaatcggg
     2821 agcgcgaacc taagctatac gaagcactga aagcacggct ggaagcacat aaagacgatc
     2881 ctgccaaagc ctttgccgag ccgttttaca aatacgataa agcaggcaac cgcacccaac
     2941 aggtaaaagc cgtacgcgta gagcaagtac agaaaaccgg cgtatgggtg cgcaaccata
     3001 acggtattgc cgacaacgca accatggtgc gcgtagatgt gtttgagaaa ggcgacaagt
     3061 attatctggt accgatttac agttggcagg tagcgaaagg gattttgccg gatagggctg
     3121 ttgtacaagg aaaagatgaa gaagattggc aacttattga tgatagtttc aactttaaat
     3181 tctcattaca ccctaatgat ttagtcgagg ttataacaaa aaaagctaga atgtttggtt
     3241 actttgccag ctgccatcga ggcacaggta atatcaatat acgcattcat gatcttgatc
     3301 ataaaattgg caaaaatgga atactggaag gtatcggcgt caaaaccgcc ctttcattcc
     3361 aaaaatacca aattgacgaa ctgggcaaag aaatcagacc atgccgtctg aaaaaacgcc
     3421 cgcctgtccg tgattataaa gatgatgatg ataaatgagc actcgagcac caccaccacc
     3481 accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     3541 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     3601 tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt
     3661 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3721 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3781 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3841 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3901 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3961 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     4021 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     4081 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4141 tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc
     4201 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     4261 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     4321 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     4381 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     4441 tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     4501 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     4561 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     4621 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     4681 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     4741 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     4801 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     4861 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     4921 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt
     4981 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     5041 tattgttcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5101 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5161 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5221 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     5281 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5341 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5401 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5461 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5521 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5581 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5641 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5701 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5761 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5821 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5881 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     5941 accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     6001 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     6061 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     6121 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     6181 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     6241 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     6301 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
     6361 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
     6421 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
     6481 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
     6541 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
     6601 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
     6661 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
     6721 ttctgctaac cagtaaggca accccgccag cctagcctgg tcctcaacga caggagcacg
     6781 atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     6841 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     6901 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     6961 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
     7021 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     7081 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     7141 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     7201 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     7261 gggcaacagc tgattgccct tcaccgcctg gccctgagat agttgcagca agcggtccac
     7321 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     7381 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
     7441 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     7501 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     7561 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     7621 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     7681 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     7741 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     7801 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     7861 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     7921 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     7981 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     8041 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     8101 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     8161 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     8221 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     8281 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
     8341 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
     8401 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     8461 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtga
//
