LOCUS       sequence-163050-        9151 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dCas9 expressed from aTc-inducible promoter in acsA, E RBS: AGGGCG.
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: pCas2E
FEATURES             Location/Qualifiers
     source          1..9151
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(64..83)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     274..295
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        510..614
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(615..683,684..1475)
                     /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(833..852)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1646..2234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2135..2154
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2388..2405
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2522..2543
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2558..2563,2564..2581,2582..2588)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2596..2612
                     /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     2601..2623
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2620..2636
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2620..2636
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2654..2672
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2654..2671
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      complement(3232..3259)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(3275..3346)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3394..3426)
                     /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(3427..4047)
                     /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     3955..3974
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     RBS             4054..4065
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(4074..4108)
                     /label=BBa_J23113
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.01"
     CDS             join(4238..4264,4265..4267,4268..6754,6755..6757,
                     6758..8344)
                     /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(8380..8398)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      8394..8441
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(8945..8964)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8946..8964)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8971..8988)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8971..8987)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8980..9002)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      join(9128..9151,1..432)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
       61 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
      121 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
      181 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
      241 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
      301 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
      361 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
      421 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
      481 catcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
      541 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
      601 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
      661 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
      721 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
      781 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
      841 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
      901 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
      961 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1021 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1081 atgtaactcg ccttgatcgt tgggaaccgg agatgaatga agccatacca aacgacgagc
     1141 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     1201 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     1261 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     1321 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     1381 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     1441 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     1501 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     1561 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     1621 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     1681 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     1741 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     1801 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     1861 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     1921 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     1981 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2041 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2101 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2161 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2221 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2281 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2341 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2401 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     2461 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     2521 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     2581 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     2641 attacgccaa gctatttagg tgacactata gaatactcaa gctatgaggt gactgccgca
     2701 ctcaatgttc aaccagatct agtgattatt tcggggttag cagccgatgc gggcaacctg
     2761 gtgaagcaac tgcgagaatt aggttacaac ggcattattg ttggggggaa cggcttaaat
     2821 acttctaata ttttccccgt ctgccaagca aaatgtgatg gggtgttggt ggcccaagcc
     2881 tacagtgccg agttagataa tgagattaac cgcgcgtttc gggacgccta ttttcaacaa
     2941 aaccaaaaag agccgcccca atttagtgcc caggctttta cggcgatcca agtttttgtt
     3001 gaagccctca gcagcctcga tgaaaaaacg cccttagaaa ctcttgctct accggacttg
     3061 cgacgacaac tgcgggacga aatttttgca ggtacctacg tcacgccttt gggtgaaatt
     3121 tccttcacag aggaagggga aattgtccag aaggaatttt ttgtggccca aattgaaatg
     3181 gatgaatcgg gtcaacaggg gcgtttcgcc ttcattgaaa cgaactataa acgcagaaag
     3241 gcccacccga aggtgagcca gtgtgactct agtagagagc gttcaccgac aaacaacaga
     3301 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggctctag
     3361 tagtgatcta cactagcact atcagtgtta ttaagctact aaagcgtagt tttcgtcgtt
     3421 tgcagcggac ccactttcac atttaagttg tttttctaat ccgcatatga tcaattcaag
     3481 gccgaataag aaggctggct ctgcaccttg gtgatcaaat aattcgatag cttgtcgtaa
     3541 taatggcggc atactatcag tagtaggtgt ttccctttct tctttagcga cttgatgctc
     3601 ttgatcttcc aatacgcaac ctaaagtaaa atgccccaca gcgctgagtg catataatgc
     3661 attctctagt gaaaaacctt gttggcataa aaaggctaat tgattttcga gagtttcata
     3721 ctgtttttct gtaggccgtg tacctaaatg tacttttgct ccatcgcgat gacttagtaa
     3781 agcacatcta aaacttttag cgttattacg taaaaaatct tgccagcttt ccccttctaa
     3841 agggcaaaag tgagtatggt gcctatctaa catctcaatg gctaaggcgt cgagcaaagc
     3901 ccgcttattt tttacatgcc aatacaatgt aggctgctct acacctagct tctgggcgag
     3961 tttacgggtt gttaaacctt cgattccgac ctcattaagc agctctaatg cgctgttaat
     4021 cactttactt ttatctaatc tggacatcta gtatttctcc tctttctcta gtagctagca
     4081 taatccctag gactgagcta gccatcaggt taacaaaaaa gcaggaatat ccctatcagt
     4141 gatagacaga catccctatc agtgatagag tataaagtta actgtgggat tgcaaaagga
     4201 gctcccaact cataaagtca agtagggcgt taattccatg gataagaaat actcaatagg
     4261 cttagctatc ggcacaaata gcgtcggatg ggcggtgatc actgatgaat ataaggttcc
     4321 gtctaaaaag ttcaaggttc tgggaaatac agaccgccac agtatcaaaa aaaatcttat
     4381 aggggctctt ttatttgaca gtggagagac agcggaagcg actcgtctca aacggacagc
     4441 tcgtagaagg tatacacgtc ggaagaatcg tatttgttat ctacaggaga ttttttcaaa
     4501 tgagatggcg aaagtagatg atagtttctt tcatcgactt gaagagtctt ttttggtgga
     4561 agaagacaag aagcatgaac gtcatcctat ttttggaaat atagtagatg aagttgctta
     4621 tcatgagaaa tatccaacta tctatcatct gcgaaaaaaa ttggtagatt ctactgataa
     4681 agcggatttg cgcttaatct atttggcctt agcgcatatg attaagtttc gtggtcattt
     4741 tttgattgag ggagatttaa atcctgataa tagtgatgtg gacaaactat ttatccagtt
     4801 ggtacaaacc tacaatcaat tatttgaaga aaaccctatt aacgcaagtg gagtagatgc
     4861 taaagcgatt ctttctgcac gattgagtaa atcaagacga ttagaaaatc tcattgctca
     4921 gctccccggt gagaagaaaa atggattatt tgggaatctc attgctttgt cattgggttt
     4981 gacccctaat tttaaatcaa attttgattt ggcagaagat gctaaattac agctttcaaa
     5041 agatacttac gatgatgatt tagataattt attggcgcaa attggagatc aatatgctga
     5101 tttgtttttg gcagctaaga atttatcaga tgctatttta ctttcagata tcctaagagt
     5161 aaatactgaa ataactaagg ctcccctatc agcttcaatg attaaacgct acgatgaaca
     5221 tcatcaagac ttgactcttt taaaagcttt agttcgacaa caacttccag aaaagtataa
     5281 agaaatcttt tttgatcaat caaaaaacgg atatgcaggt tatattgatg ggggagctag
     5341 ccaagaagaa ttttataaat ttatcaaacc aattttagaa aaaatggatg gtactgagga
     5401 attattggtg aaactaaatc gtgaagattt gctgcgcaag caacggacct ttgacaacgg
     5461 ctctattccc catcaaattc acttgggtga gctgcatgct attttgagaa gacaagaaga
     5521 cttttatcca tttttaaaag acaatcgtga gaagattgaa aaaatcttga cttttcgaat
     5581 tccttattat gttggtccat tggcgcgtgg caatagtcgt tttgcatgga tgactcggaa
     5641 gtctgaagaa acaattaccc catggaattt tgaagaagtt gtcgataaag gtgcttcagc
     5701 tcaatcattt attgaacgca tgacaaactt tgataaaaat cttccaaatg aaaaagtact
     5761 accaaaacat agtttgcttt atgagtattt tacggtttat aacgaattga caaaggtcaa
     5821 atatgttact gaaggaatgc gaaaaccagc atttctttca ggtgaacaga agaaagccat
     5881 tgttgattta ctcttcaaaa caaatcgaaa agtaaccgtt aagcaattaa aagaagatta
     5941 tttcaaaaaa atagaatgtt ttgatagtgt tgaaatttca ggagttgaag atagatttaa
     6001 tgcttcatta ggtacctacc atgatttgct aaaaattatt aaagataaag attttttgga
     6061 taatgaagaa aatgaagata tcttagagga tattgtttta acattgacct tatttgaaga
     6121 tagggagatg attgaggaaa gacttaaaac atatgctcac ctctttgatg ataaggtgat
     6181 gaaacagctt aaacgtcgcc gttatactgg ttggggacgt ttgtctcgaa aattgattaa
     6241 tggtattagg gataagcaat ctggcaaaac aatattagat tttttgaaat cagatggttt
     6301 tgccaatcgc aattttatgc agctgatcca tgatgatagt ttgacattta aagaagacat
     6361 tcaaaaagca caagtgtctg gacaaggcga tagtttacat gaacatattg caaatttagc
     6421 tggtagccct gctattaaaa aaggtatttt acagactgta aaagttgttg atgaattggt
     6481 caaagtaatg gggcggcata agccagaaaa tatcgttatt gaaatggcac gtgaaaatca
     6541 gacaactcaa aagggccaga aaaattcgcg agagcgtatg aaacgaatcg aagaaggtat
     6601 caaagaatta ggaagtcaga ttcttaaaga gcatcctgtt gaaaatactc aattgcaaaa
     6661 tgaaaagctc tatctctatt atctccaaaa tggaagagac atgtatgtgg accaagaatt
     6721 agatattaat cgtttaagtg attatgatgt cgatgccatt gttccacaaa gtttccttaa
     6781 agacgattca atagacaata aggtcttaac gcgttctgat aaaaatcgtg gtaaatcgga
     6841 taacgttcca agtgaagaag tagtcaaaaa gatgaaaaac tattggagac aacttctaaa
     6901 cgccaagtta atcactcaac gtaagtttga taatttaacg aaagctgaac gtggaggttt
     6961 gagtgaactt gataaagctg gttttatcaa acgccaattg gttgaaactc gccaaatcac
     7021 taagcatgtg gcacaaattt tggatagtcg catgaatact aaatacgatg aaaatgataa
     7081 acttattcga gaggttaaag tgattacctt aaaatctaaa ttagtttctg acttccgaaa
     7141 agatttccaa ttctataaag tacgtgagat taacaattac catcatgccc atgatgcgta
     7201 tctaaatgcc gtcgttggaa ctgctttgat taagaaatat ccaaaacttg aatcggagtt
     7261 tgtctatggt gattataaag tttatgatgt tcgtaaaatg attgctaagt ctgagcaaga
     7321 aataggcaaa gcaaccgcaa aatatttctt ttactctaat atcatgaact tcttcaaaac
     7381 agaaattaca cttgcaaatg gagagattcg caaacgccct ctaatcgaaa ctaatgggga
     7441 aactggagaa attgtctggg ataaagggcg agattttgcc acagtgcgca aagtattgtc
     7501 catgccccaa gtcaatattg tcaagaaaac agaagtacag acaggcggat tctccaagga
     7561 gtcaatttta ccaaaaagaa attcggacaa gcttattgct cgtaaaaaag actgggatcc
     7621 aaaaaaatat ggtggttttg atagtccaac ggtagcttat tcagtcctag tggttgctaa
     7681 ggtggaaaaa gggaaatcga agaagttaaa atccgttaaa gagttactag ggatcacaat
     7741 tatggaaaga agttcctttg aaaaaaatcc gattgacttt ttagaagcta aaggatataa
     7801 ggaagttaaa aaagacttaa tcattaaact acctaaatat agtctttttg agttagaaaa
     7861 cggtcgtaaa cggatgctgg ctagtgccgg agaattacaa aaaggaaatg agctggctct
     7921 gccaagcaaa tatgtgaatt ttttatattt agctagtcat tatgaaaagt tgaagggtag
     7981 tccagaagat aacgaacaaa aacaattgtt tgtggagcag cataagcatt atttagatga
     8041 gattattgag caaatcagtg aattttctaa gcgtgttatt ttagcagatg ccaatttaga
     8101 taaagttctt agtgcatata acaaacatag agacaaacca atacgtgaac aagcagaaaa
     8161 tattattcat ttatttacgt tgacgaatct tggagctccc gctgctttta aatattttga
     8221 tacaacaatt gatcgtaaac gatatacgtc tacaaaagaa gttttagatg ccactcttat
     8281 ccatcaatcc atcactggtc tttatgaaac acgcattgat ttgagtcagc taggaggtga
     8341 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     8401 aaccccttgg ggcctctaaa cgggtcttga cgggtttttt gtctagagcg ctcaagttga
     8461 tcgaatgtat gccccggata tgacctaggg agattcttct aaagcgagca gggtttgcca
     8521 agtggcgatc gcctcaggga accataacca actttgttcg aggcggcggg ggttgagctg
     8581 ggaacgatcg agatctaggg cgatcgcccg gagtttttcg gctttcccta caagaatttc
     8641 ctggtcaacg tcggtgggtt gcctttgggc caaatcgtac atactaatcg ccaggcccgc
     8701 ataaatctcc gcctggaggg ctggggggaa atcctgggtc gttgccgccg ctaaggcttc
     8761 ataccaacga tcattggcgg cttcgtaatc tgcattgagg taatggacaa aacccagggc
     8821 catcaaaatt tcggggtcgt tgggctgttg ggctagggcc atgtcccaat tgtcttgcac
     8881 cgtggcgatc gccgttgctt gggttggcga tggtgggacg gcagccaaag ttgttcagca
     8941 tgcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg
     9001 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     9061 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgtgca gcctgaatgg
     9121 cgaatggacg cgccctgtag cggcgcatta a
//
