LOCUS       sequence-135055-       10674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains dCas9 from S. pyogenes under aTc inducible promoter. 
            Suicide vector inserts into psba1 site of Synechocystis. Carries 
            spectinomycin resist. Recommend E. coli Copy cutter for 
            propogation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10674)
  AUTHORS   Yao L, Cengic I, Anfelt J, Hudson EP
  TITLE     Multiple Gene Repression in Cyanobacteria Using CRISPRi.
  JOURNAL   ACS Synth Biol. 2015 Dec 28.
  PUBMED    26689101
REFERENCE   2  (bases 1 to 10674)
  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: "ACS Synth 
            Biol. 2015 Dec 28."
COMMENT     Sequence Label: pMD19T-psba1-TetR-PL22-dCas9-SpR
FEATURES             Location/Qualifiers
     source          1..10674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(172..963,964..1032))
                     /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     795..814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1033..1137)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1205..1223
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1261..1283)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1383..1402
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1596..1618
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1610..1627
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1611..1627
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1881..1890
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2515..2536
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        2537..2571
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             2572..2583
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2590..3210
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(2663..2682)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             3211..3243
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag (LVA)
                     /note="mutant LVA variant that confers accelerated 
                     degradation under some conditions (Andersen et al., 1998)"
                     /translation="AANDENYALVA"
     terminator      3291..3362
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3378..3405
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    3420..3438
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3445..3463
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             join(3498..3524,3525..3527,3528..6014,6015..6017,
                     6018..7601)
                     /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             7602..7631
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      7651..7722
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7738..7765
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(7807..8598)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(8599..8690)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    8714..8734
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(9685..9701)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9685..9701)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9698..9720)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9709..9725
                     /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(9733..9739,9740..9757,9758..9763))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9778..9799
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9916..9933)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(10087..10674,1))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10167..10186)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
       61 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
      121 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
      181 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
      241 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
      301 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
      361 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
      421 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
      481 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
      541 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
      601 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
      661 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
      721 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
      781 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
      841 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
      901 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
      961 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     1021 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     1081 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatggggg ttccgcgcac
     1141 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     1201 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     1261 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     1321 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     1381 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     1441 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     1501 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     1561 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     1621 ggccagtgaa ttagaactcg gtacgcgcgg atcttccaga gatttatagc gttgcagtcc
     1681 ctggcgggtg agtctaactc cggcaatctg ctccgctggt accgctggaa tattgatgct
     1741 aaataaggtc ggttccgtca atggcgatcg ccagaggtgt tggacaaagc aattggcaaa
     1801 gtcggcggcg ggctgaaaat cggtggcggt aaaactgacc aaactcaaag caatgctagg
     1861 aatgccctcg atcaaccctt ccatggcggc ggaaactgta ccggaatata aaacatcagt
     1921 gccgaggtta gcaccgtggt taactccgga aaggacaaaa tccggataac ggggcatcac
     1981 cgcatggagg gcaaacttca cacagtcagc gggggtaccg gaacaggacc aagccttgat
     2041 gcgaggatca aaaacatcct ccacaatgcc cgctcgaatg ggtcgatgga gggtcagacc
     2101 atggccggtg gcggatcttt cccggtcagg acagaccact gtgacttcat ggcctgcggc
     2161 gactagggtg ttggccaagg tacgtacccc ttgggaaaaa atgccgtcat cgttactgac
     2221 aaggagattg aggaccgggg atggggtcat aggacaccaa agaatcgaga ttgtgcctgc
     2281 cagccttaac gatatcaggc tatgtcccgc ttaaactctg attcttacca ggtaactcct
     2341 gggctccatg gggccacaac caggcagtat tttgttcctt tggccaatgg ggcgatcggg
     2401 gaaaaatggc ttgatctggc atttacgaga aaaattttta ttttttaatg atttattttt
     2461 tcctattaaa atcttttttt tacctttgga aaccaactgc aatctgagaa accagaattc
     2521 gcggccgctt ctagagttta cagctagctc agtcctaggt attatgctag caaagaggag
     2581 aaatactaga tgtccagatt agataaaagt aaagtgatta acagcgcatt agagctgctt
     2641 aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg cccagaagct aggtgtagag
     2701 cagcctacat tgtattggca tgtaaaaaat aagcgggctt tgctcgacgc cttagccatt
     2761 gagatgttag ataggcacca tactcacttt tgccctttag aaggggaaag ctggcaagat
     2821 tttttacgta ataacgctaa aagttttaga tgtgctttac taagtcatcg cgatggagca
     2881 aaagtacatt taggtacacg gcctacagaa aaacagtatg aaactctcga aaatcaatta
     2941 gcctttttat gccaacaagg tttttcacta gagaatgcat tatatgcact cagcgctgtg
     3001 gggcatttta ctttaggttg cgtattggaa gatcaagagc atcaagtcgc taaagaagaa
     3061 agggaaacac ctactactga tagtatgccg ccattattac gacaagctat cgaattattt
     3121 gatcaccaag gtgcagagcc agccttctta ttcggccttg aattgatcat atgcggatta
     3181 gaaaaacaac ttaaatgtga aagtgggtcc gctgcaaacg acgaaaacta cgctttagta
     3241 gcttaataac actgatagtg ctagtgtaga tcactactag agccaggcat caaataaaac
     3301 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     3361 tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat atactagagt
     3421 ccctatcagt gatagagatt gacatcccta tcagtgatag agatactggg agctactaga
     3481 gtagtggagg ttactagatg gataagaaat actcaatagg cttagctatc ggcacaaata
     3541 gcgtcggatg ggcggtgatc actgatgaat ataaggttcc gtctaaaaag ttcaaggttc
     3601 tgggaaatac agaccgccac agtatcaaaa aaaatcttat aggggctctt ttatttgaca
     3661 gtggagagac agcggaagcg actcgtctca aacggacagc tcgtagaagg tatacacgtc
     3721 ggaagaatcg tatttgttat ctacaggaga ttttttcaaa tgagatggcg aaagtagatg
     3781 atagtttctt tcatcgactt gaagagtctt ttttggtgga agaagacaag aagcatgaac
     3841 gtcatcctat ttttggaaat atagtagatg aagttgctta tcatgagaaa tatccaacta
     3901 tctatcatct gcgaaaaaaa ttggtagatt ctactgataa agcggatttg cgcttaatct
     3961 atttggcctt agcgcatatg attaagtttc gtggtcattt tttgattgag ggagatttaa
     4021 atcctgataa tagtgatgtg gacaaactat ttatccagtt ggtacaaacc tacaatcaat
     4081 tatttgaaga aaaccctatt aacgcaagtg gagtagatgc taaagcgatt ctttctgcac
     4141 gattgagtaa atcaagacga ttagaaaatc tcattgctca gctccccggt gagaagaaaa
     4201 atggcttatt tgggaatctc attgctttgt cattgggttt gacccctaat tttaaatcaa
     4261 attttgattt ggcagaagat gctaaattac agctttcaaa agatacttac gatgatgatt
     4321 tagataattt attggcgcaa attggagatc aatatgctga tttgtttttg gcagctaaga
     4381 atttatcaga tgctatttta ctttcagata tcctaagagt aaatactgaa ataactaagg
     4441 ctcccctatc agcttcaatg attaaacgct acgatgaaca tcatcaagac ttgactcttt
     4501 taaaagcttt agttcgacaa caacttccag aaaagtataa agaaatcttt tttgatcaat
     4561 caaaaaacgg atatgcaggt tatattgatg ggggagctag ccaagaagaa ttttataaat
     4621 ttatcaaacc aattttagaa aaaatggatg gtactgagga attattggtg aaactaaatc
     4681 gtgaagattt gctgcgcaag caacggacct ttgacaacgg ctctattccc catcaaattc
     4741 acttgggtga gctgcatgct attttgagaa gacaagaaga cttttatcca tttttaaaag
     4801 acaatcgtga gaagattgaa aaaatcttga cttttcgcat tccttattat gttggtccat
     4861 tggcgcgtgg caatagtcgt tttgcatgga tgactcggaa gtctgaagaa acaattaccc
     4921 catggaattt tgaagaagtt gtcgataaag gtgcttcagc tcaatcattt attgaacgca
     4981 tgacaaactt tgataaaaat cttccaaatg aaaaagtact accaaaacat agtttgcttt
     5041 atgagtattt tacggtttat aacgaattga caaaggtcaa atatgttact gaaggaatgc
     5101 gaaaaccagc atttctttca ggtgaacaga agaaagccat tgttgattta ctcttcaaaa
     5161 caaatcgaaa agtaaccgtt aagcaattaa aagaagatta tttcaaaaaa atagaatgtt
     5221 ttgatagtgt tgaaatttca ggagttgaag atagatttaa tgcttcatta ggtacctacc
     5281 atgatttgct aaaaattatt aaagataaag attttttgga taatgaagaa aatgaagata
     5341 tcttagagga tattgtttta acattgacct tatttgaaga tagggagatg attgaggaaa
     5401 gacttaaaac atatgctcac ctctttgatg ataaggtgat gaaacagctt aaacgtcgcc
     5461 gttatactgg ttggggacgt ttgtctcgaa aattgattaa tggtattagg gataagcaat
     5521 ctggcaaaac aatattagat tttttgaaat cagatggttt tgccaatcgc aattttatgc
     5581 agctgatcca tgatgatagt ttgacattta aagaagacat tcaaaaagca caagtgtctg
     5641 gacaaggcga tagtttacat gaacatattg caaatttagc tggtagccct gctattaaaa
     5701 aaggtatttt acagactgta aaagttgttg atgaattggt caaagtaatg gggcggcata
     5761 agccagaaaa tatcgttatt gaaatggcac gtgaaaatca gacaactcaa aagggccaga
     5821 aaaattcgcg agagcgtatg aaacgaatcg aagaaggtat caaagaatta ggaagtcaga
     5881 ttcttaaaga gcatcctgtt gaaaatactc aattgcaaaa tgaaaagctc tatctctatt
     5941 atctccaaaa tggaagagac atgtatgtgg accaagaatt agatattaat cgtttaagtg
     6001 attatgatgt cgatgccatt gttccacaaa gtttccttaa agacgattca atagacaata
     6061 aggtcttaac gcgttctgat aaaaatcgtg gtaaatcgga taacgttcca agtgaagaag
     6121 tagtcaaaaa gatgaaaaac tattggagac aacttctaaa cgccaagtta atcactcaac
     6181 gtaagtttga taatttaacg aaagctgaac gtggaggttt gagtgaactt gataaagctg
     6241 gttttatcaa acgccaattg gttgaaactc gccaaatcac taagcatgtg gcacaaattt
     6301 tggatagtcg catgaatact aaatacgatg aaaatgataa acttattcga gaggttaaag
     6361 tgattacctt aaaatctaaa ttagtttctg acttccgaaa agatttccaa ttctataaag
     6421 tacgtgagat taacaattac catcatgccc atgatgcgta tctaaatgcc gtcgttggaa
     6481 ctgctttgat taagaaatat ccaaaacttg aatcggagtt tgtctatggt gattataaag
     6541 tttatgatgt tcgtaaaatg attgctaagt ctgagcaaga aataggcaaa gcaaccgcaa
     6601 aatatttctt ttactctaat atcatgaact tcttcaaaac agaaattaca cttgcaaatg
     6661 gagagattcg caaacgccct ctaatcgaaa ctaatgggga aactggagaa attgtctggg
     6721 ataaagggcg agattttgcc acagtgcgca aagtattgtc catgccccaa gtcaatattg
     6781 tcaagaaaac agaagtacag acaggcggat tctccaagga gtcaatttta ccaaaaagaa
     6841 attcggacaa gcttattgct cgtaaaaaag actgggatcc aaaaaaatat ggtggttttg
     6901 atagtccaac ggtagcttat tcagtcctag tggttgctaa ggtggaaaaa gggaaatcga
     6961 agaagttaaa atccgttaaa gagttactag ggatcacaat tatggaaaga agttcctttg
     7021 aaaaaaatcc gattgacttt ttagaagcta aaggatataa ggaagttaaa aaagacttaa
     7081 tcattaaact acctaaatat agtctttttg agttagaaaa cggtcgtaaa cggatgctgg
     7141 ctagtgccgg agaattacaa aaaggaaatg agctggctct gccaagcaaa tatgtgaatt
     7201 ttttatattt agctagtcat tatgaaaagt tgaagggtag tccagaagat aacgaacaaa
     7261 aacaattgtt tgtggagcag cataagcatt atttagatga gattattgag caaatcagtg
     7321 aattttctaa gcgtgttatt ttagcagatg ccaatttaga taaagttctt agtgcatata
     7381 acaaacatag agacaaacca atacgtgaac aagcagaaaa tattattcat ttatttacgt
     7441 tgacgaatct tggagctccc gctgctttta aatattttga tacaacaatt gatcgtaaac
     7501 gatatacgtc tacaaaagaa gttttagatg ccactcttat ccatcaatcc atcactggtc
     7561 tttatgaaac acgcattgat ttgagtcagc taggaggtga cgaacaaaaa ttgattagtg
     7621 aagaagattt gtaatactag agccaggcat caaataaaac gaaaggctca gtcgaaagac
     7681 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc
     7741 tcaccttcgg gtgggccttt ctgcgtttat atactagagc cctcggcttg aacgaattgt
     7801 tagacattat ttgccgacta ccttggtgat ctcgcctttc acgtagtgga caaattcttc
     7861 caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt ccaagataag cctgtctagc
     7921 ttcaagtatg acgggctgat actgggccgg caggcgctcc attgcccagt cggcagcgac
     7981 atccttcggc gcgattttgc cggttactgc gctgtaccaa atgcgggaca acgtaagcac
     8041 tacatttcgc tcatcgccag cccagtcggg cggcgagttc catagcgtta aggtttcatt
     8101 tagcgcctca aatagatcct gttcaggaac cggatcaaag agttcctccg ccgctggacc
     8161 taccaaggca acgctatgtt ctcttgcttt tgtcagcaag atagccagat caatgtcgat
     8221 cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc tgccattctc caaattgcag
     8281 ttcgcgctta gctggataac gccacggaat gatgtcgtcg tgcacaacaa tggtgacttc
     8341 tacagcgcgg agaatctcgc tctctccagg ggaagccgaa gtttccaaaa ggtcgttgat
     8401 caaagctcgc cgcgttgttt catcaagcct tacggtcacc gtaaccagca aatcaatatc
     8461 actgtgtggc ttcaggccgc catccactgc ggagccgtac aaatgtacgg ccagcaacgt
     8521 cggttcgaga tggcgctcga tgacgccaac tacctctgat agttgagtcg atacttcggc
     8581 gatcaccgct tccctcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8641 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa tagctagctc
     8701 actcggtcgc tactactagt agcggccgct gcagtttagc ttccggtgat gcccaaatgg
     8761 tggccctgaa cgcccccgcc attgaaggct agtgccattg ccataactgc tttcggttag
     8821 acttcgtttc atttggttaa tcaagggcac tctcgcaatg gggtgccttt tatggtccaa
     8881 ggttaaagtt aagccagtaa agtttaagtc tatttctagg gtgaaatgta atgaatcaat
     8941 ttagggactg gggtttttcc actgattggt ggcaaggcaa aaaaggggaa tattgggtac
     9001 ttgggcaaac aattctctcc gttggatttg tactgttacc ggtttatact cctactaatt
     9061 tacagttact agctgaacaa aatcagtgga ttgcctgggc tggtactctg ttttttggca
     9121 tgattgcggc agttttgctg attggggggg gtctgcattt aggggaaaat ctgactccct
     9181 tgccccatcc taaaaaggat agtcaattgg tcaccactgg tatttacagc atggtccgtc
     9241 atcccctcta cggcggcgtg gtttttctgg cgatcgccta tggcttttgg caatggagtt
     9301 tgacccattt aatcggagca gtggtgttat ttatattttt taatctcaaa gctaatcggg
     9361 aagaaatttg gctccaagat aaatttaacg actatggcaa ttatcgagac caggtgaaaa
     9421 agctgattcc ttggctttat taaaggaata tgaagaaaaa tatgggaaaa aactatgtgg
     9481 gatgaaagat ttagccagtc agaatatgtt tacggtacag aaccgaatga ttttttggtc
     9541 agtgtggcca atcaaattcc ccagggaaaa attctttgtt tagcagaagg agaaggtcgt
     9601 aatgcttgtt ttttagcaag tttgggttat gaagtgaccg cggtaatcgt cgaacggcag
     9661 gcgtgcaaac ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     9721 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     9781 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     9841 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     9901 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     9961 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
    10021 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
    10081 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
    10141 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
    10201 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
    10261 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
    10321 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
    10381 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
    10441 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
    10501 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
    10561 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
    10621 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctca
//
