LOCUS       Exported               11362 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  for cloning new sgRNAs.
ACCESSION   .
VERSION     .
KEYWORDS    pJMP1358
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11362)
  AUTHORS   Peters JM, Koo BM, Patino R, Heussler GE, Hearne CC, Qu J, Inclan 
            YF, Hawkins JS, Lu CHS, Silvis MR, Harden MM, Osadnik H, Peters JE, 
            Engel JN, Dutton RJ, Grossman AD, Gross CA, Rosenberg OS
  TITLE     Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi.
  JOURNAL   Nat Microbiol. 2019 Jan 7. pii: 10.1038/s41564-018-0327-z. doi: 
            10.1038/s41564-018-0327-z.
  PUBMED    30617347
REFERENCE   2  (bases 1 to 11362)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11362
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     mobile_element  152..317
                     /mobile_element_type="transposon:Tn7"
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     CDS             complement(472..561)
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             complement(571..4674)
                     /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"
     protein_bind    4716..4732
                     /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)."
     protein_bind    4739..4753
                     /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        5114..5191
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             5192..6274
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5211..5230)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    6538..6552
                     /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)."
     protein_bind    6559..6575
                     /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)."
     protein_bind    complement(6997..7044)
                     /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        7121..7149
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     RBS             7165..7176
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             7402..8190
                     /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"
     protein_bind    complement(8219..8266)
                     /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)."
     terminator      8317..8403
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      8506..8600
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(8628..8644)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8629..8645)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(9059..9078)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9178..9200
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9238..9256)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9324..9428
                     /gene="bla"
                     /label=AmpR promoter
     CDS             9429..10289
                     /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     complement(9647..9666)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10590..10978
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            11247..11355
                     /note="incP origin of transfer"
ORIGIN
        1 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcgg gtcagtgagc gaggaagcgg
       61 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagcg
      121 cccaaacata acaggaagaa aaatgccccg ctgtgggcgg acaaaatagt tgggaactgg
      181 gaggggtgga aatggagttt ttaaggatta tttagggaag agtgacaaaa tagatgggaa
      241 ctgggtgtag cgtcgtaagc taatacgaaa attaaaaatg acaaaatagt ttggaactag
      301 atttcactta tctggttctt gaggcggtta aaagagccgt actcttctcc gatgtcgact
      361 aggccatgat gctcattctg tgggaccaaa acgaaaaaac accctttcgg gtgtcttttc
      421 tggaatttgg taccgaggct gcaagtccca ttaaggaggc gcgccttatt acagatcttc
      481 ctcagagatg agcttctgtt ccagatcttc ttctgaaatc aacttttgtt ccagatcttc
      541 ttcagagatg agtttctgct ccgcggccgc gtcgcctccc agctgagaca ggtcgatccg
      601 tgtctcgtac aggccggtga tgctctggtg gatcagggtg gcgtccagca cctctttggt
      661 gctggtgtac ctcttccggt cgatggtggt gtcaaagtac ttgaaggcgg caggggctcc
      721 cagattggtc agggtaaaca ggtggatgat attctcggcc tgctctctga taggcttgtc
      781 tctgtgcttg ttgtaggcgc tcagcacctt gtccagatta gcgtcggcca ggatcactct
      841 cttggagaac tcgctgatct gctcgatgat ctcgtccagg tagtgtttgt gctgttccac
      901 aaacagctgt ttctgctcat tatcctcggg ggagcccttc agcttctcat agtggctggc
      961 caggtacagg aagttcacat atttggaggg cagggccagt tcgtttccct tctgcagttc
     1021 gccggcagag gccagcattc tcttccggcc gttttccagc tcgaacaggg agtacttagg
     1081 cagcttgatg atcaggtcct ttttcacttc tttgtagccc ttggcttcca gaaagtcgat
     1141 gggattcttc tcgaagctgc ttctttccat gatggtgatc cccagcagct ctttcacact
     1201 cttcagtttc ttggacttgc ccttttccac tttggccacc accagcacag aataggccac
     1261 ggtggggctg tcgaagccgc cgtacttctt agggtcccag tccttctttc tggcgatcag
     1321 cttgtcgctg ttcctcttgg gcaggataga ctctttgctg aagccgcctg tctgcacctc
     1381 ggtctttttc acgatattca cttggggcat agacagcact ttccgcacgg tggcaaagtc
     1441 ccggccctta tcccacacga tctcgcctgt ttcgccgttt gtctcgatca gaggccgctt
     1501 ccggatctcg ccgttggcca gggtaatctc ggtcttgaaa aagttcatga tgttgctgta
     1561 gaagaagtac ttggcggtag ccttgccgat ttcctgctcg ctcttggcga tcatcttccg
     1621 cacgtcgtac accttgtagt cgccgtacac gaactcgctt tccagcttag ggtacttttt
     1681 gatcagggcg gttcccacga cggcgttcag gtaggcgtcg tgggcgtggt ggtagttgtt
     1741 gatctcgcgc actttgtaaa actggaaatc cttccggaaa tcggacacca gcttggactt
     1801 cagggtgatc actttcactt cccggatcag tttgtcgttc tcgtcgtact tagtgttcat
     1861 ccgggagtcc aggatctgtg ccacgtgctt tgtgatctgc cgggtttcca ccagctgtct
     1921 cttgatgaag ccggccttat ccagttcgct caggccgcct ctctcggcct tggtcagatt
     1981 gtcgaacttc ctctgggtaa tcagcttggc attcagcagc tggcgccagt agttcttcat
     2041 cttcttcacg acctcttcgg agggcacgtt gtcgctcttg ccccggttct tgtcgctccg
     2101 agtcagcact ttgttatcga tggagtcgtc cttcagaaag ctctgaggca cgatagcgtc
     2161 cacatcgtag tcggacagcc ggttgatgtc cagttcctgg tccacgtaca tatcccgccc
     2221 attctgcagg tagtacaggt acagcttctc gttctgcagc tgggtgtttt ccacggggtg
     2281 ttctttcagg atctggctgc ccagctcttt gatgccctct tcgatccgct tcattctctc
     2341 gcggctgttc ttctgtccct tctgggtggt ctggttctct ctggccattt cgatcacgat
     2401 gttctcgggc ttgtgccggc ccatcacttt cacgagctcg tccaccacct tcactgtctg
     2461 caggatgccc ttcttaatgg cggggctgcc ggccagattg gcaatgtgct cgtgcaggct
     2521 atcgccctgg ccggacacct gggctttctg gatgtcctct ttaaaggtca ggctgtcgtc
     2581 gtggatcagc tgcatgaagt ttctgttggc gaagccgtcg gacttcagga aatccaggat
     2641 tgtcttgccg gactgcttgt cccggatgcc gttgatcagc ttccggctca gcctgcccca
     2701 gccggtgtat ctccgccgct tcagctgctt catcactttg tcgtcgaaca ggtgggcata
     2761 ggttttcagc cgttcctcga tcatctctct gtcctcaaac agtgtcaggg tcagcacgat
     2821 atcttccaga atgtcctcgt tttcctcatt gtccaggaag tccttgtcct tgataatttt
     2881 cagcagatcg tggtatgtgc ccagggaggc gttgaaccga tcttccacgc cggagatttc
     2941 cacggagtcg aagcactcga ttttcttgaa gtagtcctct ttcagctgct tcacggtcac
     3001 tttccggttg gtcttgaaca gcaggtccac gatggctttt ttctgctcgc cgctcaggaa
     3061 ggcgggcttt ctcattccct cggtcacgta tttcactttg gtcagctcgt tgtacacggt
     3121 gaagtactcg tacagcaggc tgtgcttggg cagcaccttc tcgttgggca ggttcttatc
     3181 gaagttggtc atccgctcga tgaagctctg ggcgctggcg cccttgtcca ccacttcctc
     3241 gaagttccag ggggtgatgg tttcctcgct ctttctggtc atccaggcga atctgctgtt
     3301 tcccctggcc agagggccca cgtagtaggg gatgcggaag gtcaggatct tctcgatctt
     3361 ttcccggttg tccttcagga atgggtaaaa atcttcctgc cgccgcagaa tggcgtgcag
     3421 ctctcccagg tggatctggt gggggatgct gccgttgtcg aaggtccgct gcttccgcag
     3481 caggtcctct ctgttcagct tcacgagcag ttcctcggtg ccgtccatct tttccaggat
     3541 gggcttgatg aacttgtaga actcttcctg gctggctccg ccatcgatgt agccggcgta
     3601 gccgttcttg ctctggtcga agaaaatctc tttgtacttc tcaggcagct gctgccgcac
     3661 gagagctttc agcagggtca ggtcctggtg gtgctcgtcg tatctcttga tcatagaggc
     3721 gctcaggggg gccttggtga tctcggtgtt cactctcagg atgtcgctca gcaggatggc
     3781 gtcggacagg ttcttggcgg ccagaaacag gtcggcgtac tggtcgccga tctgggccag
     3841 caggttgtcc aggtcgtcgt cgtaggtgtc cttgctcagc tgcagtttgg catcctcggc
     3901 caggtcgaag ttgctcttga agttgggggt caggcccagg ctcagggcaa tcaggttgcc
     3961 gaacaggcca ttcttcttct cgccgggcag ctgggcgatc agattttcca gccgtctgct
     4021 cttgctcagt ctggcagaca ggatggcctt ggcgtccacg ccgctggcgt tgatggggtt
     4081 ttcctcgaac agctggttgt aggtctgcac cagctggatg aacagcttgt ccacgtcgct
     4141 gttgtcgggg ttcaggtcgc cctcgatcag gaagtggccc cggaacttga tcatgtgggc
     4201 cagggccaga tagatcagcc gcaggtcggc cttgtcggtg ctgtccacca gtttctttct
     4261 caggtggtag atggtggggt acttctcgtg gtaggccacc tcgtccacga tgttgccgaa
     4321 gatggggtgc cgctcgtgct tcttatcctc ttccaccagg aaggactctt ccagtctgtg
     4381 gaagaagctg tcgtccacct tggccatctc gttgctgaag atctcttgca gatagcagat
     4441 ccggttcttc cgtctggtgt atcttcttct ggcggttctc ttcagccggg tggcctcggc
     4501 tgtttctccg ctgtcgaaca gcagggcgcc gatcaggttc ttcttgatgc tgtgccggtc
     4561 ggtgttgccc agcaccttga atttcttgct gggcaccttg tactcgtcgg tgatcacggc
     4621 ccagcccaca gagttggtgc cgatggccag gccgatgctg tacttcttgt ccatactagt
     4681 tttcctgtgt gaaacctgct gatgtgctca gtatcttgtt atccgctcac aatgtcaatg
     4741 ttatccgctc acatttataa tattttatct gattaataag atgatcttct tgagatcgtt
     4801 ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag ggcggttttt
     4861 cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttggt gataagctgt
     4921 caaaccagat caattcgcac tagtctgcag acgtaaaaaa agcggcgtgg ttagccgctt
     4981 ttttaattgc cggagcgcaa taaaaaagcc cccggaaggt gatcttccgg gggctttctc
     5041 atgcgttgct tgtattgaaa atgggggcca tgcctaccta tctgcctccc gggaagcggc
     5101 atgcatttac gttgacacca tcgaatggtg caaaaccttt cgcggtatgg catgatagcg
     5161 cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc
     5221 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
     5281 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
     5341 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
     5401 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
     5461 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
     5521 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
     5581 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
     5641 ctctgaccag acacccatca acagtattat tttctcccat gaagacggta cgcgactggg
     5701 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
     5761 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
     5821 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
     5881 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
     5941 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt
     6001 agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa
     6061 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
     6121 ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct
     6181 ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     6241 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc
     6301 tcactcatta ggcaccccac ccgggtcctt gagccccatt atcacctcca aagccttctc
     6361 gtctggtcag tttcacctgt tttacgtaaa aacccgcttc ggcgggtttt tacttttggg
     6421 gaaacacaga aaaaagcccg cacctgacag tgcgggcttt ttttttcgac caaaggtgcg
     6481 actactcttg cctactacct atcgactgag ctgaaagaat tcctaaagat ctataaatgt
     6541 gagcggataa cattgacatt gtgagcggat aacaagatac ttctagttga gaccaacttt
     6601 ggtctccacc atagcggtcg gtctctgttt aagagctatg ctggaaacag catagcaagt
     6661 ttaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttt
     6721 tgaattcatg tggctgaccg ttctgttgtc tctcgctctt ccgagtagac gaacaataag
     6781 gcctccctaa cggggggcct tttttattga taacaaaagt cagtgcttcc gctatttcca
     6841 aaataccggg ctaatacggt ttaaacgacc tcctggattt gctcagacag ccttttcgtc
     6901 attcgtttca gccaaaaaac ttaagaccgc cggtcttgtc cactaccttg cagtaatgcg
     6961 gtggacagga tcggcggttt tcttttctct tctcaagaag ttcctatact ttctagagaa
     7021 taggaacttc ggaataggaa cttcctcctg aacggccata agaacgaagg ctgtctgttg
     7081 aactctcgag ccgtggaaac ggatgaaggc acgaacccag ttgacataag cctgttcggt
     7141 tcgtaaactg taatgcagtg atctaaagag gagaaaggat ctatgcgctc acgcaactgg
     7201 tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt
     7261 tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac
     7321 gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc
     7381 cctaaaacaa agttaaacat catgagggaa gcggtgatcg ccgaagtatc gactcaacta
     7441 tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg
     7501 tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg
     7561 gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact
     7621 tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac
     7681 gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag
     7741 cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc
     7801 ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc
     7861 tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg
     7921 aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt
     7981 tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg
     8041 gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga
     8101 caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg
     8161 aaaggcgaga tcaccaaggt agtcggcaaa tctcgagcat cagcttcaaa agcgctctga
     8221 agttcctata ctttctagag aataggaact tcggaatagg tacttcaaga tccccaattc
     8281 gagctcatcg tccgggccgc aagctcctag cggcggattt gtcctactca ggagagcgtt
     8341 caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat
     8401 ttgatgcctc aagctagaga gtcattaccc caggcgttta agggcaccaa taactgcctt
     8461 aaaaaaatta cgccccgccc tgccactcat cgcagtctag cttggattct caccaataaa
     8521 aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac
     8581 tggatctatc aacaggagtc caagctcagc taattaagct agcttatcga taccgtcgac
     8641 ctcgaacccc acgcccctct ttaatacgac gggcaatttg cacttcagaa aatgaagagt
     8701 ttgctttagc cataacaaaa gtccagtatg ctttttcaca gcataactgg actgatttca
     8761 gtttacaact attctgtcta gtttaagact ttattgtcat agtttagatc tattttgttc
     8821 agtttaagac tttattgtcc gcccacacct gcttacgcag ggcatccatt tattactcaa
     8881 ccgtaaccga ttttgccagg ttacgcggct ggtcctctag ctggcgtaat agcgaagagg
     8941 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
     9001 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta
     9061 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     9121 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     9181 ggagctgcat gtgtcagagg ttttcaccgt cgtcaccgaa acgcgcgaga cgaaagggcc
     9241 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     9301 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     9361 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     9421 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     9481 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     9541 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     9601 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     9661 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     9721 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     9781 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     9841 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     9901 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     9961 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
    10021 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
    10081 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
    10141 gtgggtcccg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
    10201 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
    10261 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
    10321 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
    10381 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
    10441 aaaagatcaa aggcagttat tggtgcctca ctgattaagc attggtaact gtcagaccaa
    10501 gtttactcat atatacttta gattgattta aaacttcatt tttaattttt gcggccgcaa
    10561 gatccggcca cgatgcgtcc ggcgtagagg atctgaagat cagcagttca acctgttgat
    10621 agtacgtact aagctctcat gtttcacgta ctaagctctc atgtttaacg tactaagctc
    10681 tcatgtttaa cgaactaaac cctcatggct aacgtactaa gctctcatgg ctaacgtact
    10741 aagctctcat gtttcacgta ctaagctctc atgtttgaac aataaaatta atataaatca
    10801 gcaacttaaa tagcctctaa ggttttaagt tttataagaa aaaaaagaat atataaggct
    10861 tttaaagctt ttaaggttta acggttgtgg acaacaagcc agggatgtaa cgcactgaga
    10921 agcccttaga gcctctcaaa gcaattttga gtgacacagg aacacttaac ggctgacatg
    10981 ggaattagcc atggcatcac agtatcgtga tgacagaggc agggagtgcg gcctttttac
    11041 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcaggggatt
    11101 ccttaaggta tactttccgc tgcataaccc tgcttcgggg tcattatagc gattttttcg
    11161 gtatatccat cctttttcgc acgatataca ggattttgcc aaagggttcg tgtagacttt
    11221 ccttggtgta tccaacggcg tcagccgggc aggataggtg aagtaggccc acccgcgagc
    11281 gggtgttcct tcttcactgt cccttattcg cacctggcgg tgctcaacgg gaatcctgct
    11341 ctgcgaggct ggccgataag ct
//
