LOCUS       sequence-272930-        9714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains Promoter 1 with bsa1 cut sites for inserting sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9714)
  AUTHORS   Qu J, Prasad NK, Yu MA, Chen S, Lyden A, Herrera N, Silvis MR, 
            Crawford E, Looney MR, Peters JM, Rosenberg OS
  TITLE     Modulating pathogenesis with Mobile-CRISPRi.
  JOURNAL   J Bacteriol. 2019 Sep 3. pii: JB.00304-19. doi: 10.1128/JB.00304-19.
  PUBMED    31481541
REFERENCE   2  (bases 1 to 9714)
  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: "J 
            Bacteriol. 2019 Sep 3. pii: JB.00304-19. doi: 10.1128/JB.00304-19."
COMMENT     Sequence Label: PJQ47
FEATURES             Location/Qualifiers
     source          1..9714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     561..580
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(799..903)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     971..989
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1027..1049)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1149..1168
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1582..1598
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1583..1599
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      1627..1721
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      1824..1910
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    1961..2008
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(2056..2589)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2778..2783,2784..2800,2801..2806))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    2923..2970
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        complement(join(3386..3391,3392..3408,3409..3414))
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     promoter        3847..3881
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             3887..3898
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(3905..3931,3932..3934,3935..6421,6422..6424,
                     6425..8008)
                     /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             3905..8008
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /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             8018..8107
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     mobile_element  8262..8427
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     oriT            8586..8694
                     /note="incP origin of transfer"
     rep_origin      8963..9351
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(join(9652..9714,1..729,730..798))
                     /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"
ORIGIN
        1 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
       61 tgctgcaatg ataccgcggg acccacgctc accggctcca gatttatcag caataaacca
      121 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
      181 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
      241 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
      301 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
      361 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
      421 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
      481 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
      541 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
      601 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
      661 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
      721 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
      781 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
      841 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      901 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
      961 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgacgacgg
     1021 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     1081 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     1141 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     1201 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     1261 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctag aggaccagcc
     1321 gcgtaacctg gcaaaatcgg ttacggttga gtaataaatg gatgccctgc gtaagcaggt
     1381 gtgggcggac aataaagtct taaactgaac aaaatagatc taaactatga caataaagtc
     1441 ttaaactaga cagaatagtt gtaaactgaa atcagtccag ttatgctgtg aaaaagcata
     1501 ctggactttt gttatggcta aagcaaactc ttcattttct gaagtgcaaa ttgcccgtcg
     1561 tattaaagag gggcgtgggg ttcgaggtcg acggtatcga taagctagct taattagctg
     1621 agcttggact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca
     1681 gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagctag actgcgatga
     1741 gtggcagggc ggggcgtaat ttttttaagg cagttattgg tgcccttaaa cgcctggggt
     1801 aatgactctc tagcttgagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     1861 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgctag
     1921 gagcttgcgg cccggacgat gagctcgaat tggggatctt gaagtaccta ttccgaagtt
     1981 cctattctct agaaagtata ggaacttcag agcgcttttg aagctgatgc tcgagatctc
     2041 ggcttgaacg aattgttagg tggcggtact tgggtcgata tcaaagtgca tcacttcttc
     2101 ccgtatgccc aactttgtat agagagccac tgcgggatcg tcaccgtaat ctgcttgcac
     2161 gtagatcaca taagcaccaa gcgcgttggc ctcatgcttg aggagattga tgagcgcggt
     2221 ggcaatgccc tgcctccggt gctctccgga gactgcgaga tcatagatat agatctcact
     2281 acgcggctgc tcaaacttgg gcagaacgta agccgcgaga gcgccaacaa ccgcttcttg
     2341 gtcgaaggca gcaagcgcga tgaatgtctt actacggagc aagttcccga ggtaatcgga
     2401 gtccggctga tgttgggagt aggtggctac gtctccgaac tcacgaccga aaagatcaag
     2461 agcagcccgc atggatttga cttggtcagg gccgagccta catgtgcgaa tgatgcccat
     2521 acttgagcca cctaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct
     2581 gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa cgcgcttgct
     2641 gcttggatgc ccgaggcata gactgtacaa aaaaacagtc ataacaagcc atgaaaaccg
     2701 ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca
     2761 tacgctactt gcattacagt ttacgaaccg aacaggctta tgtcaactgg gttcgtgcct
     2821 tcatccgttt ccacggtgtg cgtccatggg caaatattat acgcaaggcg acaaggctcg
     2881 agagttcaac agacagcctt cgttcttatg gccgttcagg aggaagttcc tattccgaag
     2941 ttcctattct ctagaaagta taggaacttc ttgagaagag aaaagaaaac cgccgatcct
     3001 gtccaccgca ttactgcaag gtagtggaca agaccggcgg tcttaagttt tttggctgaa
     3061 acgaatgacg aaaaggctgt ctgagcaaat ccaggaggtc gtttaaaccg tattagcccg
     3121 gtattttgga aatagcggaa gcactgactt ttgttatcaa taaaaaaggc cccccgttag
     3181 ggaggcctta ttgttcgtct actcggaaga gcgagagaca acagaacggt cagccacatg
     3241 aattcaaaaa aaaagcaccg actcggtgcc actttttcaa gttgataacg gactagcctt
     3301 atttaaactt gctatgctgt ttccagcata gctcttaaac agagaccgac cgctatggtg
     3361 gagaccaaag ttggtctcaa ctagaattat acgagccgga tgattaattg tcaacagctc
     3421 atttcagaat atttgccaga accggaattc tttcagctca gtcgataggt agtaggcaag
     3481 agtactcgca cctttggtcg aaaaaaaaag cccgcactgt caggtgcggg cttttttctg
     3541 tgtttcccca aaagtaaaaa cccgccgaag cgggttttta cgtaaaacag gtgaaactga
     3601 ccagacgaga aggctttgga ggtgataatg gggctcaagg acccgggagg cagataggta
     3661 ggcatggccc ccattttcaa tacaagcaac gcatgagaaa gcccccggaa gatcaccttc
     3721 cgggggcttt tttattgcgc tccggcaatt aaaaaagcgg ctaaccacgc cgcttttttt
     3781 acgtctgcag actagtgcga attgatctgg tttgacagct tatcaccaag ccagttacct
     3841 cggttcttga cagctagctc agtcctaggg attgtgctag cgatctaaag aggagaaaac
     3901 tagtatggac aagaagtaca gcatcggcct ggccatcggc accaactctg tgggctgggc
     3961 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
     4021 ccggcacagc atcaagaaga acctgatcgg cgccctgctg ttcgacagcg gagaaacagc
     4081 cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat
     4141 ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca
     4201 cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt
     4261 cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag
     4321 aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc
     4381 ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag
     4441 cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa
     4501 ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag
     4561 cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg
     4621 caacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc
     4681 cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct
     4741 ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc
     4801 catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc
     4861 ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt
     4921 gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta
     4981 cgccggctac atcgatggcg gagccagcca ggaagagttc tacaagttca tcaagcccat
     5041 cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct
     5101 gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct
     5161 gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa
     5221 gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa
     5281 cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga
     5341 ggaagtggtg gacaagggcg ccagcgccca gagcttcatc gagcggatga ccaacttcga
     5401 taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac
     5461 cgtgtacaac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt
     5521 cctgagcggc gagcagaaaa aagccatcgt ggacctgctg ttcaagacca accggaaagt
     5581 gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga
     5641 aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa
     5701 aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat
     5761 cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta
     5821 tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg
     5881 gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat
     5941 cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga
     6001 cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag
     6061 cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca
     6121 gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat
     6181 cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga
     6241 gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca
     6301 ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg
     6361 gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga
     6421 cgctatcgtg cctcagagct ttctgaagga cgactccatc gataacaaag tgctgactcg
     6481 gagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat
     6541 gaagaactac tggcgccagc tgctgaatgc caagctgatt acccagagga agttcgacaa
     6601 tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag
     6661 acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat
     6721 gaacactaag tacgacgaga acgacaaact gatccgggaa gtgaaagtga tcaccctgaa
     6781 gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa
     6841 caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa
     6901 aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg
     6961 gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta
     7021 cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa
     7081 gcggcctctg atcgagacaa acggcgaaac aggcgagatc gtgtgggata agggccggga
     7141 ctttgccacc gtgcggaaag tgctgtctat gccccaagtg aatatcgtga aaaagaccga
     7201 ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgacaagct
     7261 gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt
     7321 ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag
     7381 tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat
     7441 cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc
     7501 taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga
     7561 actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc
     7621 cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt
     7681 ggaacagcac aaacactacc tggacgagat catcgagcag atcagcgagt tctccaagag
     7741 agtgatcctg gccgacgcta atctggacaa ggtgctgagc gcctacaaca agcacagaga
     7801 caagcctatc agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg
     7861 agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac
     7921 caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg
     7981 gatcgacctg tctcagctgg gaggcgacgc ggccgcggag cagaaactca tctctgaaga
     8041 agatctggaa caaaagttga tttcagaaga agatctggaa cagaagctca tctctgagga
     8101 agatctgtaa taaggcgcgc ctccttaatg ggacttgcag cctcggtacc aaattccaga
     8161 aaagacaccc gaaagggtgt tttttcgttt tggtcccaca gaatgagcat catggcctag
     8221 tcgacatcgg agaagagtac ggctctttta accgcctcaa gaaccagata agtgaaatct
     8281 agttccaaac tattttgtca tttttaattt tcgtattagc ttacgacgct acacccagtt
     8341 cccatctatt ttgtcactct tccctaaata atccttaaaa actccatttc cacccctccc
     8401 agttcccaac tattttgtcc gcccacagcg gggcattttt cttcctgtta tgtttgggcg
     8461 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     8521 gcttcctcgc tcactgaccc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagag
     8581 cttatcggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg cgaataaggg
     8641 acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct gcccggctga
     8701 cgccgttgga tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt
     8761 gcgaaaaagg atggatatac cgaaaaaatc gctataatga ccccgaagca gggttatgca
     8821 gcggaaagta taccttaagg aatcccctga taacgcagga aagaacatgt gagcaaaagg
     8881 ccagcaaaag gccaggaacc gtaaaaaggc cgcactccct gcctctgtca tcacgatact
     8941 gtgatgccat ggctaattcc catgtcagcc gttaagtgtt cctgtgtcac tcaaaattgc
     9001 tttgagaggc tctaagggct tctcagtgcg ttacatccct ggcttgttgt ccacaaccgt
     9061 taaaccttaa aagctttaaa agccttatat attctttttt ttcttataaa acttaaaacc
     9121 ttagaggcta tttaagttgc tgatttatat taattttatt gttcaaacat gagagcttag
     9181 tacgtgaaac atgagagctt agtacgttag ccatgagagc ttagtacgtt agccatgagg
     9241 gtttagttcg ttaaacatga gagcttagta cgttaaacat gagagcttag tacgtgaaac
     9301 atgagagctt agtacgtact atcaacaggt tgaactgctg atcttcagat cctctacgcc
     9361 ggacgcatcg tggccggatc ttgcggccgc aaaaattaaa aatgaagttt taaatcaatc
     9421 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacca
     9481 ataactgcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     9541 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     9601 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     9661 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagt
//
