LOCUS       sequence-200255-        9105 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express SaCas9 in bacteria with a 6xHis tag for purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9105)
  AUTHORS   Soares Medeiros LC, South L, Peng D, Bustamante JM, Wang W, 
            Bunkofske M, Perumal N, Sanchez-Valdez F, Tarleton RL
  TITLE     Rapid, Selection-Free, High-Efficiency Genome Editing in Protozoan 
            Parasites Using CRISPR-Cas9 Ribonucleoproteins.
  JOURNAL   MBio. 2017 Nov 7;8(6). pii: e01788-17. doi: 10.1128/mBio.01788-17.
  PUBMED    29114029
REFERENCE   2  (bases 1 to 9105)
  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: "MBio."; 
            date: "2017-11-7"; volume: "8(6). pii: e01788-17. doi"; pages: " 
            10.1128/mBio.01788-17"
COMMENT     Sequence Label: p6XHis_NLS-SaCas9
FEATURES             Location/Qualifiers
     source          1..9105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      28..483
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(115..134)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     325..346
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        509..613
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(614..682,683..1474)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(832..851)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1645..2233
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2134..2153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2387..2404
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2419..2558
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2484..2503)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2644..2666
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2660..2851)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3660..4742)
                     /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     4704..4723
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4743..4820)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5026..5045
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5133..5152
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5133..5151
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5152..5176
                     /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             5191..5213
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5221..5547
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             5569..5586
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5596..5613
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5620..5664
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             5680..5694
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             5698..5718
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5728..8889
                     /codon_start=1
                     /product="Cas9 endonuclease from the Staphylococcus aureus 
                     Type II CRISPR/Cas system"
                     /label=SaCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MKRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNE
                     GRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEE
                     FSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGE
                     VRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFG
                     WKDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKF
                     QIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKE
                     IIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKA
                     INLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSI
                     KVINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENA
                     KYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVK
                     QEENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRF
                     SVQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKER
                     NKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEI
                     FITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKD
                     NDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKK
                     DNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKN
                     LDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLL
                     NRIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQ
                     IIKKG"
     CDS             8965..8982
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(9035..9053)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      9049..9096
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
       61 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
      121 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
      181 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
      241 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
      301 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
      361 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
      421 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
      481 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
      541 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
      601 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
      661 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
      721 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
      781 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
      841 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
      901 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
      961 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1021 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1081 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     1141 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     1201 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     1261 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     1321 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     1381 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     1441 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     1501 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1561 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1621 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1681 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1741 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     1801 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1861 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1921 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1981 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2041 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2101 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2161 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2221 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2281 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     2341 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     2401 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2461 acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     2521 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
     2581 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     2641 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     2701 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     2761 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     2821 tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt
     2881 tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     2941 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     3001 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg
     3061 ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg
     3121 ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg
     3181 ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt
     3241 cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca
     3301 cgatcatgcg cacccgtggg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
     3361 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg
     3421 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc
     3481 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
     3541 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     3601 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     3661 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3721 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     3781 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     3841 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     3901 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc
     3961 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     4021 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     4081 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     4141 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     4201 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     4261 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     4321 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     4381 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     4441 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     4501 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     4561 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     4621 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     4681 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     4741 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     4801 ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag
     4861 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg
     4921 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
     4981 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
     5041 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
     5101 gaggatcgag atcgatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg
     5161 agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg agatatacat
     5221 atgagcgata aaattattca cctgactgac gacagttttg acacggatgt actcaaagcg
     5281 gacggggcga tcctcgtcga tttctgggca gagtggtgcg gtccgtgcaa aatgatcgcc
     5341 ccgattctgg atgaaatcgc tgacgaatat cagggcaaac tgaccgttgc aaaactgaac
     5401 atcgatcaaa accctggcac tgcgccgaaa tatggcatcc gtggtatccc gactctgctg
     5461 ctgttcaaaa acggtgaagt ggcggcaacc aaagtgggtg cactgtctaa aggtcagttg
     5521 aaagagttcc tcgacgctaa cctggccggt tctggttctg gccatatgca ccatcatcat
     5581 catcattctt ctggtctggt gccacgcggt tctggtatga aagaaaccgc tgctgctaaa
     5641 ttcgaacgcc agcacatgga cagcccagat ctgggtaccg atgacgacga caagatgccg
     5701 aagaaaaagc gcaaggtcga agcgtccatg aaaaggaact acattctggg gctggacatc
     5761 gggattacaa gcgtggggta tgggattatt gactatgaaa caagggacgt gatcgacgca
     5821 ggcgtcagac tgttcaagga ggccaacgtg gaaaacaatg agggacggag aagcaagagg
     5881 ggagccaggc gcctgaaacg acggagaagg cacagaatcc agagggtgaa gaaactgctg
     5941 ttcgattaca acctgctgac cgaccattct gagctgagtg gaattaatcc ttatgaagcc
     6001 agggtgaaag gcctgagtca gaagctgtca gaggaagagt tttccgcagc tctgctgcac
     6061 ctggctaagc gccgaggagt gcataacgtc aatgaggtgg aagaggacac cggcaacgag
     6121 ctgtctacaa aggaacagat ctcacgcaat agcaaagctc tggaagagaa gtatgtcgca
     6181 gagctacagc tggaacggct gaagaaagat ggcgaggtga gagggtcaat taataggttc
     6241 aagacaagcg actacgtcaa agaagccaag cagctgctga aagtgcagaa ggcttaccac
     6301 cagctggatc agagcttcat cgatacttat atcgacctgc tggagactcg gagaacctac
     6361 tatgagggac caggagaagg gagccccttc ggatggaaag acatcaagga atggtacgag
     6421 atgctgatgg gacattgcac ctattttcca gaagagctga gaagcgtcaa gtacgcttat
     6481 aacgcagatc tgtacaacgc cctgaatgac ctgaacaacc tggtcatcac cagggatgaa
     6541 aacgagaaac tggaatacta tgagaagttc cagatcatcg aaaacgtgtt taagcagaag
     6601 aaaaagccta cactgaaaca gattgctaag gagatcctgg tcaacgaaga ggacatcaag
     6661 ggctaccggg tgacaagcac tggaaaacca gagttcacca atctgaaagt gtatcacgat
     6721 attaaggaca tcacagcacg gaaagaaatc attgagaacg ccgaactgct ggatcagatt
     6781 gctaagatcc tgactatcta ccagagttcc gaggacatcc aggaagagct gactaacctg
     6841 aacagcgagc tgacccagga agagatcgaa cagattagta atctgaaggg gtacaccgga
     6901 acacacaacc tgtccctgaa agctatcaat ctgattctgg atgagctgtg gcatacaaac
     6961 gacaatcaga ttgcaatctt taaccggctg aagctggtac caaaaaaggt ggacctgagt
     7021 cagcagaaag agatcccaac cacactggtg gacgatttca ttctgtcacc cgtggtcaag
     7081 cggagcttca tccagagcat caaagtgatc aacgccatca tcaagaagta cggcctgccc
     7141 aatgatatca ttatcgagct ggctagggag aagaacagca aggacgcaca gaagatgatc
     7201 aatgagatgc agaaacgaaa ccggcagacc aatgaacgca ttgaagagat tatccgaact
     7261 accgggaaag agaacgcaaa gtacctgatt gaaaaaatca agctgcacga tatgcaggag
     7321 ggaaagtgtc tgtattctct ggaggccatc cccctggagg acctgctgaa caatccattc
     7381 aactacgagg tcgatcatat tatccccaga agcgtgtcct tcgacaattc ctttaacaac
     7441 aaggtgctgg tcaagcagga agagaactct aaaaagggca ataggactcc tttccagtac
     7501 ctgtctagtt cagattccaa gatctcttac gaaaccttta aaaagcacat tctgaatctg
     7561 gccaaaggaa agggccgcat cagcaagacc aaaaaggagt acctgctgga agagcgggac
     7621 atcaacagat tctccgtcca gaaggatttt attaaccgga atctggtgga cacaagatac
     7681 gctactcgcg gcctgatgaa tctgctgcga tcctatttcc gggtgaacaa tctggatgtg
     7741 aaagtcaagt ccatcaacgg cgggttcaca tcttttctga ggcgcaaatg gaagtttaaa
     7801 aaggagcgca acaaagggta caagcaccat gccgaagatg ctctgattat cgcaaatgcc
     7861 gacttcatct ttaaggagtg gaaaaagctg gacaaagcca agaaagtgat ggagaaccag
     7921 atgttcgaag agaagcaggc cgaatctatg cccgaaatcg agacagaaca ggagtacaag
     7981 gagattttca tcactcctca ccagatcaag catatcaagg atttcaagga ctacaagtac
     8041 tctcaccggg tggataaaaa gcccaacaga gagctgatca atgacaccct gtatagtaca
     8101 agaaaagacg ataaggggaa taccctgatt gtgaacaatc tgaacggact gtacgacaaa
     8161 gataatgaca agctgaaaaa gctgatcaac aaaagtcccg agaagctgct gatgtaccac
     8221 catgatcctc agacatatca gaaactgaag ctgattatgg agcagtacgg cgacgagaag
     8281 aacccactgt ataagtacta tgaagagact gggaactacc tgaccaagta tagcaaaaag
     8341 gataatggcc ccgtgatcaa gaagatcaag tactatggga acaagctgaa tgcccatctg
     8401 gacatcacag acgattaccc taacagtcgc aacaaggtgg tcaagctgtc actgaagcca
     8461 tacagattcg atgtctatct ggacaacggc gtgtataaat ttgtgactgt caagaatctg
     8521 gatgtcatca aaaaggagaa ctactatgaa gtgaatagca agtgctacga agaggctaaa
     8581 aagctgaaaa agattagcaa ccaggcagag ttcatcgcct ccttttacaa caacgacctg
     8641 attaagatca atggcgaact gtatagggtc atcggggtga acaatgatct gctgaaccgc
     8701 attgaagtga atatgattga catcacttac cgagagtatc tggaaaacat gaatgataag
     8761 cgcccccctc gaattatcaa aacaatcgcc tctaagactc agagtatcaa aaagtactca
     8821 accgacattc tgggaaacct gtatgaggtg aagagcaaaa agcaccctca gattatcaaa
     8881 aagggctaac cgggcttctc ctcaaccatg gcgatatcgg atccgaattc gagctccgtc
     8941 gacaagcttg cggccgcact cgagcaccac caccaccacc actgagatcc ggctgctaac
     9001 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
     9061 cttggggcct ctaaacgggt cttgaggggt tttttgctga aagga
//
