LOCUS       sequence-162353-        9151 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dCas9 expressed from aTc-inducible promoter in acsA, B RBS: TCGAGA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9151)
  AUTHORS   Gordon GC, Korosh TC, Cameron JC, Markley AL, Begemann MB, Pfleger 
            BF
  TITLE     CRISPR interference as a titratable, trans-acting regulatory tool 
            for metabolic engineering in the cyanobacterium Synechococcus sp. 
            strain PCC 7002.
  JOURNAL   Metab Eng. 2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 
            10.1016/j.ymben.2016.07.007.
  PUBMED    27481676
REFERENCE   2  (bases 1 to 9151)
  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: "Metab Eng. 
            2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 
            10.1016/j.ymben.2016.07.007."
COMMENT     Sequence Label: pCas2B
FEATURES             Location/Qualifiers
     source          1..9151
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      145..733
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     634..653
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     887..904
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1021..1042
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1057..1062,1063..1080,1081..1087)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1095..1111
                     /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)."
     primer_bind     1100..1122
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1119..1135
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1119..1135
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1153..1171
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1153..1170
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      complement(1731..1758)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(1774..1845)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1893..1925)
                     /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"
     CDS             complement(1926..2546)
                     /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     2454..2473
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     RBS             2553..2564
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(2573..2607)
                     /label=BBa_J23113
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.01"
     CDS             join(2737..2763,2764..2766,2767..5253,5254..5256,
                     5257..6843)
                     /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"
     primer_bind     complement(6879..6897)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6893..6940
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(7444..7463)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7445..7463)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7470..7487)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7470..7486)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7479..7501)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7627..8082
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7714..7733)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7924..7945
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8160..8264
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8265..8333,8334..9125)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPEMNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8483..8502)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
       61 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
      121 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
      181 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
      241 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
      301 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
      361 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
      421 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
      481 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
      541 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
      601 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
      661 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
      721 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
      781 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
      841 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
      901 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
      961 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     1021 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     1081 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     1141 ttacgccaag ctatttaggt gacactatag aatactcaag ctatgaggtg actgccgcac
     1201 tcaatgttca accagatcta gtgattattt cggggttagc agccgatgcg ggcaacctgg
     1261 tgaagcaact gcgagaatta ggttacaacg gcattattgt tggggggaac ggcttaaata
     1321 cttctaatat tttccccgtc tgccaagcaa aatgtgatgg ggtgttggtg gcccaagcct
     1381 acagtgccga gttagataat gagattaacc gcgcgtttcg ggacgcctat tttcaacaaa
     1441 accaaaaaga gccgccccaa tttagtgccc aggcttttac ggcgatccaa gtttttgttg
     1501 aagccctcag cagcctcgat gaaaaaacgc ccttagaaac tcttgctcta ccggacttgc
     1561 gacgacaact gcgggacgaa atttttgcag gtacctacgt cacgcctttg ggtgaaattt
     1621 ccttcacaga ggaaggggaa attgtccaga aggaattttt tgtggcccaa attgaaatgg
     1681 atgaatcggg tcaacagggg cgtttcgcct tcattgaaac gaactataaa cgcagaaagg
     1741 cccacccgaa ggtgagccag tgtgactcta gtagagagcg ttcaccgaca aacaacagat
     1801 aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tggctctagt
     1861 agtgatctac actagcacta tcagtgttat taagctacta aagcgtagtt ttcgtcgttt
     1921 gcagcggacc cactttcaca tttaagttgt ttttctaatc cgcatatgat caattcaagg
     1981 ccgaataaga aggctggctc tgcaccttgg tgatcaaata attcgatagc ttgtcgtaat
     2041 aatggcggca tactatcagt agtaggtgtt tccctttctt ctttagcgac ttgatgctct
     2101 tgatcttcca atacgcaacc taaagtaaaa tgccccacag cgctgagtgc atataatgca
     2161 ttctctagtg aaaaaccttg ttggcataaa aaggctaatt gattttcgag agtttcatac
     2221 tgtttttctg taggccgtgt acctaaatgt acttttgctc catcgcgatg acttagtaaa
     2281 gcacatctaa aacttttagc gttattacgt aaaaaatctt gccagctttc cccttctaaa
     2341 gggcaaaagt gagtatggtg cctatctaac atctcaatgg ctaaggcgtc gagcaaagcc
     2401 cgcttatttt ttacatgcca atacaatgta ggctgctcta cacctagctt ctgggcgagt
     2461 ttacgggttg ttaaaccttc gattccgacc tcattaagca gctctaatgc gctgttaatc
     2521 actttacttt tatctaatct ggacatctag tatttctcct ctttctctag tagctagcat
     2581 aatccctagg actgagctag ccatcaggtt aacaaaaaag caggaatatc cctatcagtg
     2641 atagacagac atccctatca gtgatagagt ataaagttaa ctgtgggatt gcaaaaggag
     2701 ctcccaactc ataaagtcaa gttcgacatt aattccatgg ataagaaata ctcaataggc
     2761 ttagctatcg gcacaaatag cgtcggatgg gcggtgatca ctgatgaata taaggttccg
     2821 tctaaaaagt tcaaggttct gggaaataca gaccgccaca gtatcaaaaa aaatcttata
     2881 ggggctcttt tatttgacag tggagagaca gcggaagcga ctcgtctcaa acggacagct
     2941 cgtagaaggt atacacgtcg gaagaatcgt atttgttatc tacaggagat tttttcaaat
     3001 gagatggcga aagtagatga tagtttcttt catcgacttg aagagtcttt tttggtggaa
     3061 gaagacaaga agcatgaacg tcatcctatt tttggaaata tagtagatga agttgcttat
     3121 catgagaaat atccaactat ctatcatctg cgaaaaaaat tggtagattc tactgataaa
     3181 gcggatttgc gcttaatcta tttggcctta gcgcatatga ttaagtttcg tggtcatttt
     3241 ttgattgagg gagatttaaa tcctgataat agtgatgtgg acaaactatt tatccagttg
     3301 gtacaaacct acaatcaatt atttgaagaa aaccctatta acgcaagtgg agtagatgct
     3361 aaagcgattc tttctgcacg attgagtaaa tcaagacgat tagaaaatct cattgctcag
     3421 ctccccggtg agaagaaaaa tggattattt gggaatctca ttgctttgtc attgggtttg
     3481 acccctaatt ttaaatcaaa ttttgatttg gcagaagatg ctaaattaca gctttcaaaa
     3541 gatacttacg atgatgattt agataattta ttggcgcaaa ttggagatca atatgctgat
     3601 ttgtttttgg cagctaagaa tttatcagat gctattttac tttcagatat cctaagagta
     3661 aatactgaaa taactaaggc tcccctatca gcttcaatga ttaaacgcta cgatgaacat
     3721 catcaagact tgactctttt aaaagcttta gttcgacaac aacttccaga aaagtataaa
     3781 gaaatctttt ttgatcaatc aaaaaacgga tatgcaggtt atattgatgg gggagctagc
     3841 caagaagaat tttataaatt tatcaaacca attttagaaa aaatggatgg tactgaggaa
     3901 ttattggtga aactaaatcg tgaagatttg ctgcgcaagc aacggacctt tgacaacggc
     3961 tctattcccc atcaaattca cttgggtgag ctgcatgcta ttttgagaag acaagaagac
     4021 ttttatccat ttttaaaaga caatcgtgag aagattgaaa aaatcttgac ttttcgaatt
     4081 ccttattatg ttggtccatt ggcgcgtggc aatagtcgtt ttgcatggat gactcggaag
     4141 tctgaagaaa caattacccc atggaatttt gaagaagttg tcgataaagg tgcttcagct
     4201 caatcattta ttgaacgcat gacaaacttt gataaaaatc ttccaaatga aaaagtacta
     4261 ccaaaacata gtttgcttta tgagtatttt acggtttata acgaattgac aaaggtcaaa
     4321 tatgttactg aaggaatgcg aaaaccagca tttctttcag gtgaacagaa gaaagccatt
     4381 gttgatttac tcttcaaaac aaatcgaaaa gtaaccgtta agcaattaaa agaagattat
     4441 ttcaaaaaaa tagaatgttt tgatagtgtt gaaatttcag gagttgaaga tagatttaat
     4501 gcttcattag gtacctacca tgatttgcta aaaattatta aagataaaga ttttttggat
     4561 aatgaagaaa atgaagatat cttagaggat attgttttaa cattgacctt atttgaagat
     4621 agggagatga ttgaggaaag acttaaaaca tatgctcacc tctttgatga taaggtgatg
     4681 aaacagctta aacgtcgccg ttatactggt tggggacgtt tgtctcgaaa attgattaat
     4741 ggtattaggg ataagcaatc tggcaaaaca atattagatt ttttgaaatc agatggtttt
     4801 gccaatcgca attttatgca gctgatccat gatgatagtt tgacatttaa agaagacatt
     4861 caaaaagcac aagtgtctgg acaaggcgat agtttacatg aacatattgc aaatttagct
     4921 ggtagccctg ctattaaaaa aggtatttta cagactgtaa aagttgttga tgaattggtc
     4981 aaagtaatgg ggcggcataa gccagaaaat atcgttattg aaatggcacg tgaaaatcag
     5041 acaactcaaa agggccagaa aaattcgcga gagcgtatga aacgaatcga agaaggtatc
     5101 aaagaattag gaagtcagat tcttaaagag catcctgttg aaaatactca attgcaaaat
     5161 gaaaagctct atctctatta tctccaaaat ggaagagaca tgtatgtgga ccaagaatta
     5221 gatattaatc gtttaagtga ttatgatgtc gatgccattg ttccacaaag tttccttaaa
     5281 gacgattcaa tagacaataa ggtcttaacg cgttctgata aaaatcgtgg taaatcggat
     5341 aacgttccaa gtgaagaagt agtcaaaaag atgaaaaact attggagaca acttctaaac
     5401 gccaagttaa tcactcaacg taagtttgat aatttaacga aagctgaacg tggaggtttg
     5461 agtgaacttg ataaagctgg ttttatcaaa cgccaattgg ttgaaactcg ccaaatcact
     5521 aagcatgtgg cacaaatttt ggatagtcgc atgaatacta aatacgatga aaatgataaa
     5581 cttattcgag aggttaaagt gattacctta aaatctaaat tagtttctga cttccgaaaa
     5641 gatttccaat tctataaagt acgtgagatt aacaattacc atcatgccca tgatgcgtat
     5701 ctaaatgccg tcgttggaac tgctttgatt aagaaatatc caaaacttga atcggagttt
     5761 gtctatggtg attataaagt ttatgatgtt cgtaaaatga ttgctaagtc tgagcaagaa
     5821 ataggcaaag caaccgcaaa atatttcttt tactctaata tcatgaactt cttcaaaaca
     5881 gaaattacac ttgcaaatgg agagattcgc aaacgccctc taatcgaaac taatggggaa
     5941 actggagaaa ttgtctggga taaagggcga gattttgcca cagtgcgcaa agtattgtcc
     6001 atgccccaag tcaatattgt caagaaaaca gaagtacaga caggcggatt ctccaaggag
     6061 tcaattttac caaaaagaaa ttcggacaag cttattgctc gtaaaaaaga ctgggatcca
     6121 aaaaaatatg gtggttttga tagtccaacg gtagcttatt cagtcctagt ggttgctaag
     6181 gtggaaaaag ggaaatcgaa gaagttaaaa tccgttaaag agttactagg gatcacaatt
     6241 atggaaagaa gttcctttga aaaaaatccg attgactttt tagaagctaa aggatataag
     6301 gaagttaaaa aagacttaat cattaaacta cctaaatata gtctttttga gttagaaaac
     6361 ggtcgtaaac ggatgctggc tagtgccgga gaattacaaa aaggaaatga gctggctctg
     6421 ccaagcaaat atgtgaattt tttatattta gctagtcatt atgaaaagtt gaagggtagt
     6481 ccagaagata acgaacaaaa acaattgttt gtggagcagc ataagcatta tttagatgag
     6541 attattgagc aaatcagtga attttctaag cgtgttattt tagcagatgc caatttagat
     6601 aaagttctta gtgcatataa caaacataga gacaaaccaa tacgtgaaca agcagaaaat
     6661 attattcatt tatttacgtt gacgaatctt ggagctcccg ctgcttttaa atattttgat
     6721 acaacaattg atcgtaaacg atatacgtct acaaaagaag ttttagatgc cactcttatc
     6781 catcaatcca tcactggtct ttatgaaaca cgcattgatt tgagtcagct aggaggtgac
     6841 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     6901 accccttggg gcctctaaac gggtcttgac gggttttttg tctagagcgc tcaagttgat
     6961 cgaatgtatg ccccggatat gacctaggga gattcttcta aagcgagcag ggtttgccaa
     7021 gtggcgatcg cctcagggaa ccataaccaa ctttgttcga ggcggcgggg gttgagctgg
     7081 gaacgatcga gatctagggc gatcgcccgg agtttttcgg ctttccctac aagaatttcc
     7141 tggtcaacgt cggtgggttg cctttgggcc aaatcgtaca tactaatcgc caggcccgca
     7201 taaatctccg cctggagggc tggggggaaa tcctgggtcg ttgccgccgc taaggcttca
     7261 taccaacgat cattggcggc ttcgtaatct gcattgaggt aatggacaaa acccagggcc
     7321 atcaaaattt cggggtcgtt gggctgttgg gctagggcca tgtcccaatt gtcttgcacc
     7381 gtggcgatcg ccgttgcttg ggttggcgat ggtgggacgg cagccaaagt tgttcagcat
     7441 gcgccctata gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg
     7501 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     7561 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgtgcag cctgaatggc
     7621 gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     7681 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     7741 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     7801 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     7861 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     7921 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     7981 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     8041 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
     8101 ccttacgcat ctgtgcggta tttcacaccg catcaggtgg cacttttcgg ggaaatgtgc
     8161 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     8221 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     8281 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     8341 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     8401 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     8461 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     8521 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     8581 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     8641 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     8701 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     8761 agatgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     8821 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     8881 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     8941 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     9001 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     9061 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     9121 ggtaactgtc agaccaagtt tactcatata t
//
