LOCUS       sequence-210275-        8508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  D16A 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
FEATURES             Location/Qualifiers
     source          1..8508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     93..112
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     196..215
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        196..214
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    215..239
                     /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             254..276
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             283..3528
                     /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
                     RLNEKGYVEIDHALPFSRTWDDSFNNKVLVLGSENQNKGNQTPYEYFNGKDNSREWQEF
                     KARVETSRFPRSKKQRILLQKFDEDGFKERNLNDTRYVNRFLCQFVADRMRLTGKGKKR
                     VFASNGQITNLLRGFWGLRKVRAENDRHHALDAVVVACSTVAMQQKITRFVRYKEMNAF
                     DGKTIDKETGEVLHQKTHFPQPWEFFAQEVMIRVFGKPDGKPEFEEADTLEKLRTLLAE
                     KLSSRPEAVHEYVTPLFVSRAPNRKMSGQGHMETVKSAKRLDEGVSVLRVPLTQLKLKD
                     LEKMVNREREPKLYEALKARLEAHKDDPAKAFAEPFYKYDKAGNRTQQVKAVRVEQVQK
                     TGVWVRNHNGIADNATMVRVDVFEKGDKYYLVPIYSWQVAKGILPDRAVVQGKDEEDWQ
                     LIDDSFNFKFSLHPNDLVEVITKKARMFGYFASCHRGTGNINIRIHDLDHKIGKNGILE
                     GIGVKTALSFQKYQIDELGKEIRPCRLKKRPPVR"
     CDS             3529..3552
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3565..3585
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     primer_bind     complement(3638..3656)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3652..3699
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3736..4191
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3823..3842)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4033..4054
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4283..5098)
                     /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(4283..5098)
                     /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     5006..5025
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5090..5109
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5220..5808
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5709..5728
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5962..5979
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5994..6133
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6059..6078)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(6235..6426)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(7235..8317)
                     /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     8279..8298
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8318..8395)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
       61 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
      121 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc
      181 tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
      241 ctctagaaat aattttgttt aactttaaga aggagatata ccatggctgc cttcaaacct
      301 aattcaatca actacatcct cggcctcgct atcggcatcg catccgtcgg ctgggcgatg
      361 gtagaaattg acgaagaaga aaaccccatc cgcctgattg atttgggcgt gcgcgtattt
      421 gagcgtgccg aagtaccgaa aacaggcgac tcccttgcca tggcaaggcg tttggcgcgc
      481 agtgttcgcc gcctgacccg ccgtcgcgcc caccgcctgc ttcggacccg ccgcctattg
      541 aaacgcgaag gcgtattaca agccgccaat tttgacgaaa acggcttgat taaatcctta
      601 ccgaatacac catggcaact tcgcgcagcc gcattagacc gcaaactgac gcctttagag
      661 tggtcggcag tcttgttgca tttaatcaaa catcgcggct atttatcgca acggaaaaac
      721 gagggcgaaa ctgccgataa ggagcttggc gctttgctta aaggcgtagc cggcaatgcc
      781 catgccttac agacaggcga tttccgcaca ccggccgaat tggctttaaa taaatttgag
      841 aaagaaagcg gccatatccg caatcagcgc agcgattatt cgcatacgtt cagccgcaaa
      901 gatttacagg cggagctgat tttgctgttt gaaaaacaaa aagaatttgg caatccgcat
      961 gtttcaggcg gccttaaaga aggtattgaa accctactga tgacgcaacg ccctgccctg
     1021 tccggcgatg ccgttcaaaa aatgttgggg cattgcacct tcgaaccggc agagccgaaa
     1081 gccgctaaaa acacctacac agccgaacgt ttcatctggc tgaccaagct gaacaacctg
     1141 cgtattttag agcaaggcag cgagcggcca ttgaccgata ccgaacgcgc cacgcttatg
     1201 gacgagccat acagaaaatc caaactgact tacgcacaag cccgtaagct gctgggttta
     1261 gaagataccg cctttttcaa aggcttgcgc tatggtaaag acaatgccga agcctcaaca
     1321 ttgatggaaa tgaaggccta ccatgccatc agccgtgcac tggaaaaaga aggattgaaa
     1381 gacaaaaaat ccccattaaa cctttctccc gaattacaag acgaaatcgg cacggcattc
     1441 tccctgttca aaaccgatga agacattaca ggccgtctga aagaccgtat acagcccgaa
     1501 atcttagaag cgctgttgaa acacatcagc ttcgataagt tcgtccaaat ttccttgaaa
     1561 gcattgcgcc gaattgtgcc tctaatggaa caaggcaaac gttacgatga agcctgcgcc
     1621 gaaatctacg gagaccatta cggcaagaag aatacggaag aaaagattta tctgccgccg
     1681 attcccgccg acgaaatccg caaccccgtc gtcttgcgcg ccttatctca agcacgtaag
     1741 gtcattaacg gcgtggtacg ccgttacggc tccccagctc gtatccatat tgaaactgca
     1801 agggaagtag gtaaatcgtt taaagaccgc aaagaaattg agaaacgcca agaagaaaac
     1861 cgcaaagacc gggaaaaagc cgccgccaaa ttccgagagt atttccccaa ttttgtcgga
     1921 gaacccaaat ccaaagatat tctgaaactg cgcctgtacg agcaacaaca cggcaaatgc
     1981 ctgtattcgg gcaaagaaat caacttaggc cgtctgaacg aaaaaggcta tgtcgaaatc
     2041 gaccatgccc tgccgttctc gcgcacatgg gacgacagtt tcaacaataa agtactggta
     2101 ttgggcagcg aaaaccaaaa caaaggcaat caaacccctt acgaatactt caacggcaaa
     2161 gacaacagcc gcgaatggca ggaatttaaa gcgcgtgtcg aaaccagccg tttcccgcgc
     2221 agtaaaaaac aacggattct gctgcaaaaa ttcgatgaag acggctttaa agaacgcaat
     2281 ctgaacgaca cgcgctacgt caaccgtttc ctgtgtcaat ttgttgccga ccgtatgcgg
     2341 ctgacaggta aaggcaagaa acgtgtcttt gcatccaacg gacaaattac caatctgttg
     2401 cgcggctttt ggggattgcg caaagtgcgt gcggaaaacg accgccatca cgccttggac
     2461 gccgtcgtcg ttgcctgctc gaccgttgcc atgcagcaga aaattacccg ttttgtacgc
     2521 tataaagaga tgaacgcgtt tgacggtaaa accatagaca aagaaacagg agaagtgctg
     2581 catcaaaaaa cacacttccc acaaccttgg gaatttttcg cacaagaagt catgattcgc
     2641 gtcttcggca aaccggacgg caaacccgaa ttcgaagaag ccgataccct agaaaaactg
     2701 cgcacgttgc ttgccgaaaa attatcatct cgccccgaag ccgtacacga atacgttacg
     2761 ccactgtttg tttcacgcgc gcccaatcgg aagatgagcg ggcaagggca tatggagacc
     2821 gtcaaatccg ccaaacgact ggacgaaggc gtcagcgtgt tgcgcgtacc gctgacacag
     2881 ttaaaactga aagacttgga aaaaatggtc aatcgggagc gcgaacctaa gctatacgaa
     2941 gcactgaaag cacggctgga agcacataaa gacgatcctg ccaaagcctt tgccgagccg
     3001 ttttacaaat acgataaagc aggcaaccgc acccaacagg taaaagccgt acgcgtagag
     3061 caagtacaga aaaccggcgt atgggtgcgc aaccataacg gtattgccga caacgcaacc
     3121 atggtgcgcg tagatgtgtt tgagaaaggc gacaagtatt atctggtacc gatttacagt
     3181 tggcaggtag cgaaagggat tttgccggat agggctgttg tacaaggaaa agatgaagaa
     3241 gattggcaac ttattgatga tagtttcaac tttaaattct cattacaccc taatgattta
     3301 gtcgaggtta taacaaaaaa agctagaatg tttggttact ttgccagctg ccatcgaggc
     3361 acaggtaata tcaatatacg cattcatgat cttgatcata aaattggcaa aaatggaata
     3421 ctggaaggta tcggcgtcaa aaccgccctt tcattccaaa aataccaaat tgacgaactg
     3481 ggcaaagaaa tcagaccatg ccgtctgaaa aaacgcccgc ctgtccgtga ttataaagat
     3541 gatgatgata aatgagcact cgagcaccac caccaccacc accactgaga tccggctgct
     3601 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     3661 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     3721 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3781 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3841 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3901 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3961 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4021 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     4081 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     4141 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     4201 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4261 gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt
     4321 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     4381 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     4441 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     4501 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     4561 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     4621 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     4681 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     4741 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     4801 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     4861 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     4921 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     4981 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     5041 ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa
     5101 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct
     5161 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5221 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5281 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5341 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     5401 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5461 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5521 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5581 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5641 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5701 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5761 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5821 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5881 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     5941 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     6001 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     6061 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     6121 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     6181 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggatctgca tgtgtcagag
     6241 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     6301 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     6361 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     6421 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     6481 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     6541 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     6601 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     6661 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     6721 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     6781 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     6841 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
     6901 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
     6961 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
     7021 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
     7081 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
     7141 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
     7201 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7261 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7321 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
     7381 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
     7441 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
     7501 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
     7561 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
     7621 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
     7681 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
     7741 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
     7801 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
     7861 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
     7921 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
     7981 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
     8041 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
     8101 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
     8161 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     8221 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     8281 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     8341 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     8401 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     8461 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcg
//
