LOCUS       sequence-162347-        9151 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dCas9 expressed from aTc-inducible promoter in acsA, F RBS: TGGGCG.
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: pCas2F
FEATURES             Location/Qualifiers
     source          1..9151
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(493..1284,1285..1353))
                     /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     1116..1135
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1354..1458)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1536..1991)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1673..1694)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1885..1904
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2117..2139
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2131..2148
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2132..2148
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2155..2174
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2155..2173
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2678..2725)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2721..2739
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(join(2775..4361,4362..4364,4365..6851,6852..6854,
                     6855..6881))
                     /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"
     promoter        7011..7045
                     /label=BBa_J23113
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.01"
     RBS             7054..7065
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             7072..7692
                     /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(7145..7164)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             7693..7725
                     /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      7773..7844
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7860..7887
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        complement(8447..8465)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(8448..8465)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(8483..8499)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8483..8499)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8496..8518)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8507..8523
                     /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(8531..8537,8538..8555,8556..8561))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8576..8597
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8714..8731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(8885..9151,1..322))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8965..8984)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
       61 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
      121 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
      181 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
      241 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
      301 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
      361 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
      421 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
      481 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
      541 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
      601 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
      661 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
      721 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
      781 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
      841 tatggcttca ttcatctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
      901 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
      961 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     1021 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     1081 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     1141 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     1201 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     1261 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     1321 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     1381 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     1441 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgatgcgg tgtgaaatac
     1501 cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt
     1561 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     1621 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     1681 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     1741 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     1801 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     1861 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     1921 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     1981 acagggcgcg tccattcgcc attcaggctg cacaactgtt gggaagggcg atcggtgcgg
     2041 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     2101 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac
     2161 gactcactat agggcgcatg ctgaacaact ttggctgccg tcccaccatc gccaacccaa
     2221 gcaacggcga tcgccacggt gcaagacaat tgggacatgg ccctagccca acagcccaac
     2281 gaccccgaaa ttttgatggc cctgggtttt gtccattacc tcaatgcaga ttacgaagcc
     2341 gccaatgatc gttggtatga agccttagcg gcggcaacga cccaggattt ccccccagcc
     2401 ctccaggcgg agatttatgc gggcctggcg attagtatgt acgatttggc ccaaaggcaa
     2461 cccaccgacg ttgaccagga aattcttgta gggaaagccg aaaaactccg ggcgatcgcc
     2521 ctagatctcg atcgttccca gctcaacccc cgccgcctcg aacaaagttg gttatggttc
     2581 cctgaggcga tcgccacttg gcaaaccctg ctcgctttag aagaatctcc ctaggtcata
     2641 tccggggcat acattcgatc aacttgagcg ctctagacaa aaaacccgtc aagacccgtt
     2701 tagaggcccc aaggggttat gctagttatt gctcagcggt ggcagcagcc aactcagctt
     2761 cctttcgggc tttgttagtc acctcctagc tgactcaaat caatgcgtgt ttcataaaga
     2821 ccagtgatgg attgatggat aagagtggca tctaaaactt cttttgtaga cgtatatcgt
     2881 ttacgatcaa ttgttgtatc aaaatattta aaagcagcgg gagctccaag attcgtcaac
     2941 gtaaataaat gaataatatt ttctgcttgt tcacgtattg gtttgtctct atgtttgtta
     3001 tatgcactaa gaactttatc taaattggca tctgctaaaa taacacgctt agaaaattca
     3061 ctgatttgct caataatctc atctaaataa tgcttatgct gctccacaaa caattgtttt
     3121 tgttcgttat cttctggact acccttcaac ttttcataat gactagctaa atataaaaaa
     3181 ttcacatatt tgcttggcag agccagctca tttccttttt gtaattctcc ggcactagcc
     3241 agcatccgtt tacgaccgtt ttctaactca aaaagactat atttaggtag tttaatgatt
     3301 aagtcttttt taacttcctt atatccttta gcttctaaaa agtcaatcgg atttttttca
     3361 aaggaacttc tttccataat tgtgatccct agtaactctt taacggattt taacttcttc
     3421 gatttccctt tttccacctt agcaaccact aggactgaat aagctaccgt tggactatca
     3481 aaaccaccat atttttttgg atcccagtct tttttacgag caataagctt gtccgaattt
     3541 ctttttggta aaattgactc cttggagaat ccgcctgtct gtacttctgt tttcttgaca
     3601 atattgactt ggggcatgga caatactttg cgcactgtgg caaaatctcg ccctttatcc
     3661 cagacaattt ctccagtttc cccattagtt tcgattagag ggcgtttgcg aatctctcca
     3721 tttgcaagtg taatttctgt tttgaagaag ttcatgatat tagagtaaaa gaaatatttt
     3781 gcggttgctt tgcctatttc ttgctcagac ttagcaatca ttttacgaac atcataaact
     3841 ttataatcac catagacaaa ctccgattca agttttggat atttcttaat caaagcagtt
     3901 ccaacgacgg catttagata cgcatcatgg gcatgatggt aattgttaat ctcacgtact
     3961 ttatagaatt ggaaatcttt tcggaagtca gaaactaatt tagattttaa ggtaatcact
     4021 ttaacctctc gaataagttt atcattttca tcgtatttag tattcatgcg actatccaaa
     4081 atttgtgcca catgcttagt gatttggcga gtttcaacca attggcgttt gataaaacca
     4141 gctttatcaa gttcactcaa acctccacgt tcagctttcg ttaaattatc aaacttacgt
     4201 tgagtgatta acttggcgtt tagaagttgt ctccaatagt ttttcatctt tttgactact
     4261 tcttcacttg gaacgttatc cgatttacca cgatttttat cagaacgcgt taagacctta
     4321 ttgtctattg aatcgtcttt aaggaaactt tgtggaacaa tggcatcgac atcataatca
     4381 cttaaacgat taatatctaa ttcttggtcc acatacatgt ctcttccatt ttggagataa
     4441 tagagataga gcttttcatt ttgcaattga gtattttcaa caggatgctc tttaagaatc
     4501 tgacttccta attctttgat accttcttcg attcgtttca tacgctctcg cgaatttttc
     4561 tggccctttt gagttgtctg attttcacgt gccatttcaa taacgatatt ttctggctta
     4621 tgccgcccca ttactttgac caattcatca acaactttta cagtctgtaa aatacctttt
     4681 ttaatagcag ggctaccagc taaatttgca atatgttcat gtaaactatc gccttgtcca
     4741 gacacttgtg ctttttgaat gtcttcttta aatgtcaaac tatcatcatg gatcagctgc
     4801 ataaaattgc gattggcaaa accatctgat ttcaaaaaat ctaatattgt tttgccagat
     4861 tgcttatccc taataccatt aatcaatttt cgagacaaac gtccccaacc agtataacgg
     4921 cgacgtttaa gctgtttcat caccttatca tcaaagaggt gagcatatgt tttaagtctt
     4981 tcctcaatca tctccctatc ttcaaataag gtcaatgtta aaacaatatc ctctaagata
     5041 tcttcatttt cttcattatc caaaaaatct ttatctttaa taatttttag caaatcatgg
     5101 taggtaccta atgaagcatt aaatctatct tcaactcctg aaatttcaac actatcaaaa
     5161 cattctattt ttttgaaata atcttctttt aattgcttaa cggttacttt tcgatttgtt
     5221 ttgaagagta aatcaacaat ggctttcttc tgttcacctg aaagaaatgc tggttttcgc
     5281 attccttcag taacatattt gacctttgtc aattcgttat aaaccgtaaa atactcataa
     5341 agcaaactat gttttggtag tactttttca tttggaagat ttttatcaaa gtttgtcatg
     5401 cgttcaataa atgattgagc tgaagcacct ttatcgacaa cttcttcaaa attccatggg
     5461 gtaattgttt cttcagactt ccgagtcatc catgcaaaac gactattgcc acgcgccaat
     5521 ggaccaacat aataaggaat tcgaaaagtc aagatttttt caatcttctc acgattgtct
     5581 tttaaaaatg gataaaagtc ttcttgtctt ctcaaaatag catgcagctc acccaagtga
     5641 atttgatggg gaatagagcc gttgtcaaag gtccgttgct tgcgcagcaa atcttcacga
     5701 tttagtttca ccaataattc ctcagtacca tccatttttt ctaaaattgg tttgataaat
     5761 ttataaaatt cttcttggct agctccccca tcaatataac ctgcatatcc gttttttgat
     5821 tgatcaaaaa agatttcttt atacttttct ggaagttgtt gtcgaactaa agcttttaaa
     5881 agagtcaagt cttgatgatg ttcatcgtag cgtttaatca ttgaagctga taggggagcc
     5941 ttagttattt cagtatttac tcttaggata tctgaaagta aaatagcatc tgataaattc
     6001 ttagctgcca aaaacaaatc agcatattga tctccaattt gcgccaataa attatctaaa
     6061 tcatcatcgt aagtatcttt tgaaagctgt aatttagcat cttctgccaa atcaaaattt
     6121 gatttaaaat taggggtcaa acccaatgac aaagcaatga gattcccaaa taatccattt
     6181 ttcttctcac cggggagctg agcaatgaga ttttctaatc gtcttgattt actcaatcgt
     6241 gcagaaagaa tcgctttagc atctactcca cttgcgttaa tagggttttc ttcaaataat
     6301 tgattgtagg tttgtaccaa ctggataaat agtttgtcca catcactatt atcaggattt
     6361 aaatctccct caatcaaaaa atgaccacga aacttaatca tatgcgctaa ggccaaatag
     6421 attaagcgca aatccgcttt atcagtagaa tctaccaatt tttttcgcag atgatagata
     6481 gttggatatt tctcatgata agcaacttca tctactatat ttccaaaaat aggatgacgt
     6541 tcatgcttct tgtcttcttc caccaaaaaa gactcttcaa gtcgatgaaa gaaactatca
     6601 tctactttcg ccatctcatt tgaaaaaatc tcctgtagat aacaaatacg attcttccga
     6661 cgtgtatacc ttctacgagc tgtccgtttg agacgagtcg cttccgctgt ctctccactg
     6721 tcaaataaaa gagcccctat aagatttttt ttgatactgt ggcggtctgt atttcccaga
     6781 accttgaact ttttagacgg aaccttatat tcatcagtga tcaccgccca tccgacgcta
     6841 tttgtgccga tagctaagcc tattgagtat ttcttatcca tggaattaac gcccaacttg
     6901 actttatgag ttgggagctc cttttgcaat cccacagtta actttatact ctatcactga
     6961 tagggatgtc tgtctatcac tgatagggat attcctgctt ttttgttaac ctgatggcta
     7021 gctcagtcct agggattatg ctagctacta gagaaagagg agaaatacta gatgtccaga
     7081 ttagataaaa gtaaagtgat taacagcgca ttagagctgc ttaatgaggt cggaatcgaa
     7141 ggtttaacaa cccgtaaact cgcccagaag ctaggtgtag agcagcctac attgtattgg
     7201 catgtaaaaa ataagcgggc tttgctcgac gccttagcca ttgagatgtt agataggcac
     7261 catactcact tttgcccttt agaaggggaa agctggcaag attttttacg taataacgct
     7321 aaaagtttta gatgtgcttt actaagtcat cgcgatggag caaaagtaca tttaggtaca
     7381 cggcctacag aaaaacagta tgaaactctc gaaaatcaat tagccttttt atgccaacaa
     7441 ggtttttcac tagagaatgc attatatgca ctcagcgctg tggggcattt tactttaggt
     7501 tgcgtattgg aagatcaaga gcatcaagtc gctaaagaag aaagggaaac acctactact
     7561 gatagtatgc cgccattatt acgacaagct atcgaattat ttgatcacca aggtgcagag
     7621 ccagccttct tattcggcct tgaattgatc atatgcggat tagaaaaaca acttaaatgt
     7681 gaaagtgggt ccgctgcaaa cgacgaaaac tacgctttag tagcttaata acactgatag
     7741 tgctagtgta gatcactact agagccaggc atcaaataaa acgaaaggct cagtcgaaag
     7801 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg
     7861 gctcaccttc gggtgggcct ttctgcgttt atagttcgtt tcaatgaagg cgaaacgccc
     7921 ctgttgaccc gattcatcca tttcaatttg ggccacaaaa aattccttct ggacaatttc
     7981 cccttcctct gtgaaggaaa tttcacccaa aggcgtgacg taggtacctg caaaaatttc
     8041 gtcccgcagt tgtcgtcgca agtccggtag agcaagagtt tctaagggcg ttttttcatc
     8101 gaggctgctg agggcttcaa caaaaacttg gatcgccgta aaagcctggg cactaaattg
     8161 gggcggctct ttttggtttt gttgaaaata ggcgtcccga aacgcgcggt taatctcatt
     8221 atctaactcg gcactgtagg cttgggccac caacacccca tcacattttg cttggcagac
     8281 ggggaaaata ttagaagtat ttaagccgtt ccccccaaca ataatgccgt tgtaacctaa
     8341 ttctcgcagt tgcttcacca ggttgcccgc atcggctgct aaccccgaaa taatcactag
     8401 atctggttga acattgagtg cggcagtcac ctcatagctt gagtattcta tagtgtcacc
     8461 taaatagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     8521 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     8581 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     8641 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     8701 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     8761 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     8821 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     8881 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     8941 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     9001 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     9061 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     9121 gctccaagct gggctgtgtg cacgaacccc c
//
