LOCUS       sequence-357940-        9541 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of dCas9-VP64-6xHis in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9541)
  AUTHORS   Zuris JA, Thompson DB, Shu Y, Guilinger JP, Bessen JL, Hu JH, Maeder
            ML, Joung JK, Chen ZY, Liu DR
  TITLE     Cationic lipid-mediated delivery of proteins enables efficient 
            protein-based genome editing in vitro and in vivo.
  JOURNAL   Nat Biotechnol. 2015 Jan;33(1):73-80. doi: 10.1038/nbt.3081. Epub 
            2014 Oct 30.
  PUBMED    25357182
REFERENCE   2  (bases 1 to 9541)
  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: "10.1038/nbt.3081"; 
            journalName: "Nat Biotechnol"; date: "2015-01"; volume: "33"; issue:
            "1"; pages: "73-80"
COMMENT     Sequence Label: pET-dCas9-VP64-6xHis
FEATURES             Location/Qualifiers
     source          1..9541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     921..940
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(960..1037)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1243..1262
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1350..1369
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1350..1368
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1369..1393
                     /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             1408..1430
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(1437..1463,1464..1466,1467..3953,3954..3956,
                     3957..5540)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5547..5567
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5574..5639
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5670..5819
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             5820..5837
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5856..5900
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(5938..5956)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5952..5999
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      6036..6491
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6123..6142)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6333..6354
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(6606..7397,7398..7466))
                     /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     7229..7248
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7467..7559)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7640..8228
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8129..8148
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8382..8399
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8414..8553
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8479..8498)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8639..8661
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8655..8846)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(9418..9541,1..959))
                     /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"
ORIGIN
        1 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
       61 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
      121 gagctgtctt cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg
      181 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
      241 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
      301 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
      361 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
      421 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
      481 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
      541 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
      601 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
      661 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
      721 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
      781 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
      841 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
      901 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
      961 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
     1021 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
     1081 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa
     1141 ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
     1201 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
     1261 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
     1321 gatcgagatc gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc
     1381 ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga tataccatgg
     1441 ataagaaata ctcaataggc ttagctatcg gcacaaatag cgtcggatgg gcggtgatca
     1501 ctgatgaata taaggttccg tctaaaaagt tcaaggttct gggaaataca gaccgccaca
     1561 gtatcaaaaa aaatcttata ggggctcttt tatttgacag tggagagaca gcggaagcga
     1621 ctcgtctcaa acggacagct cgtagaaggt atacacgtcg gaagaatcgt atttgttatc
     1681 tacaggagat tttttcaaat gagatggcga aagtagatga tagtttcttt catcgacttg
     1741 aagagtcttt tttggtggaa gaagacaaga agcatgaacg tcatcctatt tttggaaata
     1801 tagtagatga agttgcttat catgagaaat atccaactat ctatcatctg cgaaaaaaat
     1861 tggtagattc tactgataaa gcggatttgc gcttaatcta tttggcctta gcgcatatga
     1921 ttaagtttcg tggtcatttt ttgattgagg gagatttaaa tcctgataat agtgatgtgg
     1981 acaaactatt tatccagttg gtacaaacct acaatcaatt atttgaagaa aaccctatta
     2041 acgcaagtgg agtagatgct aaagcgattc tttctgcacg attgagtaaa tcaagacgat
     2101 tagaaaatct cattgctcag ctccccggtg agaagaaaaa tggcttattt gggaatctca
     2161 ttgctttgtc attgggtttg acccctaatt ttaaatcaaa ttttgatttg gcagaagatg
     2221 ctaaattaca gctttcaaaa gatacttacg atgatgattt agataattta ttggcgcaaa
     2281 ttggagatca atatgctgat ttgtttttgg cagctaagaa tttatcagat gctattttac
     2341 tttcagatat cctaagagta aatactgaaa taactaaggc tcccctatca gcttcaatga
     2401 ttaaacgcta cgatgaacat catcaagact tgactctttt aaaagcttta gttcgacaac
     2461 aacttccaga aaagtataaa gaaatctttt ttgatcaatc aaaaaacgga tatgcaggtt
     2521 atattgatgg gggagctagc caagaagaat tttataaatt tatcaaacca attttagaaa
     2581 aaatggatgg tactgaggaa ttattggtga aactaaatcg tgaagatttg ctgcgcaagc
     2641 aacggacctt tgacaacggc tctattcccc atcaaattca cttgggtgag ctgcatgcta
     2701 ttttgagaag acaagaagac ttttatccat ttttaaaaga caatcgtgag aagattgaaa
     2761 aaatcttgac ttttcgaatt ccttattatg ttggtccatt ggcgcgtggc aatagtcgtt
     2821 ttgcatggat gactcggaag tctgaagaaa caattacccc atggaatttt gaagaagttg
     2881 tcgataaagg tgcttcagct caatcattta ttgaacgcat gacaaacttt gataaaaatc
     2941 ttccaaatga aaaagtacta ccaaaacata gtttgcttta tgagtatttt acggtttata
     3001 acgaattgac aaaggtcaaa tatgttactg aaggaatgcg aaaaccagca tttctttcag
     3061 gtgaacagaa gaaagccatt gttgatttac tcttcaaaac aaatcgaaaa gtaaccgtta
     3121 agcaattaaa agaagattat ttcaaaaaaa tagaatgttt tgatagtgtt gaaatttcag
     3181 gagttgaaga tagatttaat gcttcattag gtacctacca tgatttgcta aaaattatta
     3241 aagataaaga ttttttggat aatgaagaaa atgaagatat cttagaggat attgttttaa
     3301 cattgacctt atttgaagat agggagatga ttgaggaaag acttaaaaca tatgctcacc
     3361 tctttgatga taaggtgatg aaacagctta aacgtcgccg ttatactggt tggggacgtt
     3421 tgtctcgaaa attgattaat ggtattaggg ataagcaatc tggcaaaaca atattagatt
     3481 ttttgaaatc agatggtttt gccaatcgca attttatgca gctgatccat gatgatagtt
     3541 tgacatttaa agaagacatt caaaaagcac aagtgtctgg acaaggcgat agtttacatg
     3601 aacatattgc aaatttagct ggtagccctg ctattaaaaa aggtatttta cagactgtaa
     3661 aagttgttga tgaattggtc aaagtaatgg ggcggcataa gccagaaaat atcgttattg
     3721 aaatggcacg tgaaaatcag acaactcaaa agggccagaa aaattcgcga gagcgtatga
     3781 aacgaatcga agaaggtatc aaagaattag gaagtcagat tcttaaagag catcctgttg
     3841 aaaatactca attgcaaaat gaaaagctct atctctatta tctccaaaat ggaagagaca
     3901 tgtatgtgga ccaagaatta gatattaatc gtttaagtga ttatgatgtc gatgccattg
     3961 ttccacaaag tttccttaaa gacgattcaa tagacaataa ggtcttaacg cgttctgata
     4021 aaaatcgtgg taaatcggat aacgttccaa gtgaagaagt agtcaaaaag atgaaaaact
     4081 attggagaca acttctaaac gccaagttaa tcactcaacg taagtttgat aatttaacga
     4141 aagctgaacg tggaggtttg agtgaacttg ataaagctgg ttttatcaaa cgccaattgg
     4201 ttgaaactcg ccaaatcact aagcatgtgg cacaaatttt ggatagtcgc atgaatacta
     4261 aatacgatga aaatgataaa cttattcgag aggttaaagt gattacctta aaatctaaat
     4321 tagtttctga cttccgaaaa gatttccaat tctataaagt acgtgagatt aacaattacc
     4381 atcatgccca tgatgcgtat ctaaatgccg tcgttggaac tgctttgatt aagaaatatc
     4441 caaaacttga atcggagttt gtctatggtg attataaagt ttatgatgtt cgtaaaatga
     4501 ttgctaagtc tgagcaagaa ataggcaaag caaccgcaaa atatttcttt tactctaata
     4561 tcatgaactt cttcaaaaca gaaattacac ttgcaaatgg agagattcgc aaacgccctc
     4621 taatcgaaac taatggggaa actggagaaa ttgtctggga taaagggcga gattttgcca
     4681 cagtgcgcaa agtattgtcc atgccccaag tcaatattgt caagaaaaca gaagtacaga
     4741 caggcggatt ctccaaggag tcaattttac caaaaagaaa ttcggacaag cttattgctc
     4801 gtaaaaaaga ctgggatcca aaaaaatatg gtggttttga tagtccaacg gtagcttatt
     4861 cagtcctagt ggttgctaag gtggaaaaag ggaaatcgaa gaagttaaaa tccgttaaag
     4921 agttactagg gatcacaatt atggaaagaa gttcctttga aaaaaatccg attgactttt
     4981 tagaagctaa aggatataag gaagttaaaa aagacttaat cattaaacta cctaaatata
     5041 gtctttttga gttagaaaac ggtcgtaaac ggatgctggc tagtgccgga gaattacaaa
     5101 aaggaaatga gctggctctg ccaagcaaat atgtgaattt tttatattta gctagtcatt
     5161 atgaaaagtt gaagggtagt ccagaagata acgaacaaaa acaattgttt gtggagcagc
     5221 ataagcatta tttagatgag attattgagc aaatcagtga attttctaag cgtgttattt
     5281 tagcagatgc caatttagat aaagttctta gtgcatataa caaacataga gacaaaccaa
     5341 tacgtgaaca agcagaaaat attattcatt tatttacgtt gacgaatctt ggagctcccg
     5401 ctgcttttaa atattttgat acaacaattg atcgtaaacg atatacgtct acaaaagaag
     5461 ttttagatgc cactcttatc catcaatcca tcactggtct ttatgaaaca cgcattgatt
     5521 tgagtcagct aggaggtgac ggttctccca agaagaagag gaaagtctcg agcgactaca
     5581 aagaccatga cggtgattat aaagatcatg acatcgatta caaggatgac gatgacaagg
     5641 ctgcaggagg cggtggaagc gggcgcgccg acgcgctgga cgatttcgat ctcgacatgc
     5701 tgggttctga tgccctcgat gactttgacc tggatatgtt gggaagcgac gcattggatg
     5761 actttgatct ggacatgctc ggctccgatg ctctggacga tttcgatctc gatatgttac
     5821 atcaccacca ccatcactaa tgactcgagt ctggtaaaga aaccgctgct gcgaaatttg
     5881 aacgccagca catggactcg tctactagcg cagcttaatt aacctaggct gctgccaccg
     5941 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     6001 tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag
     6061 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     6121 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     6181 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     6241 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     6301 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     6361 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     6421 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     6481 gtttacaatt tctggcggca cgatggcatg agattatcaa aaaggatctt cacctagatc
     6541 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6601 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6661 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6721 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6781 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6841 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6901 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6961 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     7021 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     7081 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     7141 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     7201 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     7261 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     7321 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     7381 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7441 gcgacacgga aatgttgaat actcatactc ttcctttttc aatcatgatt gaagcattta
     7501 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7561 aggtcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     7621 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     7681 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     7741 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     7801 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     7861 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     7921 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     7981 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     8041 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     8101 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     8161 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     8221 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     8281 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     8341 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     8401 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     8461 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     8521 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     8581 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     8641 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     8701 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     8761 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     8821 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     8881 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     8941 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     9001 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     9061 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     9121 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     9181 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     9241 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     9301 atgctagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     9361 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     9421 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     9481 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     9541 g
//
