LOCUS       sequence-314817-       11761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Z. mobilis CRISPRi, promoter C, empty guide, sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11761)
  AUTHORS   Banta AB, Enright AL, Siletti C, Peters JM
  TITLE     A High-Efficacy CRISPR Interference System for Gene Function 
            Discovery in Zymomonas mobilis.
  JOURNAL   Appl Environ Microbiol. 2020 Nov 10;86(23). pii: AEM.01621-20. doi: 
            10.1128/AEM.01621-20. Print 2020 Nov 10.
  PUBMED    32978126
REFERENCE   2  (bases 1 to 11761)
  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: "Appl 
            Environ Microbiol."; date: "2020-11-10"; volume: "86(23). pii: 
            AEM.01621-20. doi"; pages: " 10.1128/AEM.01621-20. Print 2020 Nov 
            10"
COMMENT     Sequence Label: pJMP2367
FEATURES             Location/Qualifiers
     source          1..11761
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..78
                     /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             79..1161
                     /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(98..117)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    1437..1456
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     CDS             join(1830..1832,1833..2546)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1830..2543
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1860..1888)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2483..2503
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    complement(2688..2735)
                     /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        join(2812..2817,2818..2834,2835..2840)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     primer_bind     complement(2989..3008)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    complement(3672..3719)
                     /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      complement(3770..3856)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(3959..4053)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(4081..4097)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4082..4098)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4512..4531)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4631..4653
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4691..4709)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4777..4881
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4882..4950,4951..5742)
                     /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(5100..5119)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6043..6431
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            6700..6808
                     /note="incP origin of transfer"
     mobile_element  6967..7132
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     CDS             complement(7287..7376)
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             complement(join(7386..8969,8970..8972,8973..11459,
                     11460..11462,11463..11489))
                     /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    11512..11528
                     /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    11545..11561
                     /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)."
ORIGIN
        1 gacaccatcg aatggtgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
       61 caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt
      121 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
      181 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa
      241 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
      301 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
      361 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
      421 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
      481 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
      541 cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg
      601 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg
      661 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
      721 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
      781 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
      841 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
      901 gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
      961 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
     1021 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
     1081 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     1141 cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc
     1201 accccacccg ggtccttgag ccccattatc acctccaaag ccttctcgtc tggtcagttt
     1261 cacctgtttt acgtaaaaac ccgcttcggc gggtttttac ttttggggaa acacagaaaa
     1321 aagcccgcac ctgacagtgc gggctttttt tttcgaccaa aggtgcgact actcttgcct
     1381 actacctatc gactgagctg aaagaattcg gaaaattttt tttcaaaagt acttgaaatt
     1441 gtgagcgctc acaattataa ttctagtaga gaccaacttt ggtctccacc atagcggtcg
     1501 gtctctgttt aagagctatg ctggaaacag catagcaagt ttaaataagg ctagtccgtt
     1561 atcaacttga aaaagtggca ccgagtcggt gctttttttt tgaattcatg tggctgaccg
     1621 ttctgttgtc tctcgctctt ccgagtagac gaacaataag gcctccctaa cggggggcct
     1681 tttttattga taacaaaagt cagtgcttcc gctatttcca aaataccggg ctaatacggt
     1741 ttaaacgaaa atttatcaaa aagagtattg acttaaagtc taacctatag gatacttaca
     1801 gccagatctg agcgagaagg aggtaaagta tgagcaaagg agaagaactt ttcactggag
     1861 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtccgtg
     1921 gagagggtga aggtgatgct acaaacggaa aactcaccct taaatttatt tgcactactg
     1981 gaaaactacc tgttccgtgg ccaacacttg tcactactct gacctatggt gttcaatgct
     2041 tttcccgtta tccggatcac atgaaacggc atgacttttt caagagtgcc atgcccgaag
     2101 gttatgtaca ggaacgcact atatctttca aagatgacgg gacctacaag acgcgtgctg
     2161 aagtcaagtt tgaaggtgat acccttgtta atcgtatcga gttaaagggt attgatttta
     2221 aagaagatgg aaacattctt ggacacaaac tggagtacaa ctttaactca cacaatgtat
     2281 acatcacggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt cgccacaacg
     2341 ttgaagatgg ttccgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg
     2401 gccctgtcct tttaccagac aaccattacc tgtcgacaca atctgtcctt tcgaaagatc
     2461 ccaacgaaaa gcgtgaccac atggtccttc ttgagtttgt aactgctgct gggattacac
     2521 atggcatgga tgagctctac aaataataag tttaaacgac ctcctggatt tgctcagaca
     2581 gccttttcgt cattcgtttc agccaaaaaa cttaagaccg ccggtcttgt ccactacctt
     2641 gcagtaatgc ggtggacagg atcggcggtt ttcttttctc ttctcaagaa gttcctatac
     2701 tttctagaga ataggaactt cggaatagga acttcctcct gaacggccat aagaacgaag
     2761 gctgtctgtt gaactctcga gccgtggaaa cggatgaagg cacgaaccca gttgacataa
     2821 gcctgttcgg ttcgtaaact gtaatgcaag tagcgtatgc gctcatcact accgggcgta
     2881 ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact
     2941 ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag
     3001 tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag
     3061 accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg
     3121 atgaatgctc atccggagtt ccgtatggca atgaaagacg gtgagctggt gatatgggat
     3181 agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg
     3241 agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt
     3301 tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca
     3361 gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc
     3421 ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg
     3481 ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat
     3541 gaattacaac agtttttatg catgcgccca atacgcaaac cgcctctccc cgcgcgttgg
     3601 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaactcgag catcagcttc
     3661 aaaagcgctc tgaagttcct atactttcta gagaatagga acttcggaat aggtacttca
     3721 agatccccaa ttcgagctca tcgtccgggc cgcaagctcc tagcggcgga tttgtcctac
     3781 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct ttcgactgag
     3841 cctttcgttt tatttgatgc ctcaagctag agagtcatta ccccaggcgt ttaagggcac
     3901 caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcagtc tagcttggat
     3961 tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt
     4021 ctgaggtcat tactggatct atcaacagga gtccaagctc agctaattaa gctagcttat
     4081 cgataccgtc gacctcgaac cccacgcccc tctttaatac gacgggcaat ttgcacttca
     4141 gaaaatgaag agtttgcttt agccataaca aaagtccagt atgctttttc acagcataac
     4201 tggactgatt tcagtttaca actattctgt ctagtttaag actttattgt catagtttag
     4261 atctattttg ttcagtttaa gactttattg tccgcccaca cctgcttacg cagggcatcc
     4321 atttattact caaccgtaac cgattttgcc aggttacgcg gctggtcctc tagctggcgt
     4381 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     4441 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     4501 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4561 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4621 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcgtcacc gaaacgcgcg
     4681 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4741 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4801 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4861 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4921 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4981 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5041 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5101 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5161 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5221 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5281 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5341 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5401 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5461 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5521 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5581 ggagccggtg agcgtgggtc ccgcggtatc attgcagcac tggggccaga tggtaagccc
     5641 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5701 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5761 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5821 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5881 tcagaccccg tagaaaagat caaaggcagt tattggtgcc tcactgatta agcattggta
     5941 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     6001 tttgcggccg caagatccgg ccacgatgcg tccggcgtag aggatctgaa gatcagcagt
     6061 tcaacctgtt gatagtacgt actaagctct catgtttcac gtactaagct ctcatgttta
     6121 acgtactaag ctctcatgtt taacgaacta aaccctcatg gctaacgtac taagctctca
     6181 tggctaacgt actaagctct catgtttcac gtactaagct ctcatgtttg aacaataaaa
     6241 ttaatataaa tcagcaactt aaatagcctc taaggtttta agttttataa gaaaaaaaag
     6301 aatatataag gcttttaaag cttttaaggt ttaacggttg tggacaacaa gccagggatg
     6361 taacgcactg agaagccctt agagcctctc aaagcaattt tgagtgacac aggaacactt
     6421 aacggctgac atgggaatta gccatggcat cacagtatcg tgatgacaga ggcagggagt
     6481 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6541 ttatcagggg attccttaag gtatactttc cgctgcataa ccctgcttcg gggtcattat
     6601 agcgattttt tcggtatatc catccttttt cgcacgatat acaggatttt gccaaagggt
     6661 tcgtgtagac tttccttggt gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg
     6721 cccacccgcg agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa
     6781 cgggaatcct gctctgcgag gctggccgat aagctctgat accgctcgcc gcagccgaac
     6841 gaccgagcgc agcgggtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     6901 tctccccgcg cgttggccga ttcattaatg cagcgcccaa acataacagg aagaaaaatg
     6961 ccccgctgtg ggcggacaaa atagttggga actgggaggg gtggaaatgg agtttttaag
     7021 gattatttag ggaagagtga caaaatagat gggaactggg tgtagcgtcg taagctaata
     7081 cgaaaattaa aaatgacaaa atagtttgga actagatttc acttatctgg ttcttgaggc
     7141 ggttaaaaga gccgtactct tctccgatgt cgactaggcc atgatgctca ttctgtggga
     7201 ccaaaacgaa aaaacaccct ttcgggtgtc ttttctggaa tttggtaccg aggctgcaag
     7261 tcccattaag gaggcgcgcc ttattacaga tcttcctcag agatgagctt ctgttccaga
     7321 tcttcttctg aaatcaactt ttgttccaga tcttcttcag agatgagttt ctgctccgcg
     7381 gccgcgtcac ctcctagctg actcaaatca atgcgtgttt cataaagacc agtgatggat
     7441 tgatggataa gagtggcatc taaaacttct tttgtagacg tatatcgttt acgatcaatt
     7501 gttgtatcaa aatatttaaa agcagcggga gctccaagat tcgtcaacgt aaataaatga
     7561 ataatatttt ctgcttgttc acgtattggt ttgtctctat gtttgttata tgcactaaga
     7621 actttatcta aattggcatc tgctaaaata acacgcttag aaaattcact gatttgctca
     7681 ataatctcat ctaaataatg cttatgctgc tccacaaaca attgtttttg ttcgttatct
     7741 tctggactac ccttcaactt ttcataatga ctagctaaat ataaaaaatt cacatatttg
     7801 cttggcagag ccagctcatt tcctttttgt aattctccgg cactagccag catccgttta
     7861 cgaccgtttt ctaactcaaa aagactatat ttaggtagtt taatgattaa gtctttttta
     7921 acttccttat atcctttagc ttctaaaaag tcaatcggat ttttttcaaa ggaacttctt
     7981 tccataattg tgatccctag taactcttta acggatttta acttcttcga tttccctttt
     8041 tccaccttag caaccactag gactgaataa gctaccgttg gactatcaaa accaccatat
     8101 ttttttggat cccagtcttt tttacgagca ataagcttgt ccgaatttct ttttggtaaa
     8161 attgactcct tggagaatcc gcctgtctgt acttctgttt tcttgacaat attgacttgg
     8221 ggcatggaca atactttgcg cactgtggca aaatctcgcc ctttatccca gacaatttct
     8281 ccagtttccc cattagtttc gattagaggg cgtttgcgaa tctctccatt tgcaagtgta
     8341 atttctgttt tgaagaagtt catgatatta gagtaaaaga aatattttgc ggttgctttg
     8401 cctatttctt gctcagactt agcaatcatt ttacgaacat cataaacttt ataatcacca
     8461 tagacaaact ccgattcaag ttttggatat ttcttaatca aagcagttcc aacgacggca
     8521 tttagatacg catcatgggc atgatggtaa ttgttaatct cacgtacttt atagaattgg
     8581 aaatcttttc ggaagtcaga aactaattta gattttaagg taatcacttt aacctctcga
     8641 ataagtttat cattttcatc gtatttagta ttcatgcgac tatccaaaat ttgtgccaca
     8701 tgcttagtga tttggcgagt ttcaaccaat tggcgtttga taaaaccagc tttatcaagt
     8761 tcactcaaac ctccacgttc agctttcgtt aaattatcaa acttacgttg agtgattaac
     8821 ttggcgttta gaagttgtct ccaatagttt ttcatctttt tgactacttc ttcacttgga
     8881 acgttatccg atttaccacg atttttatca gaacgcgtta agaccttatt gtctattgaa
     8941 tcgtctttaa ggaaactttg tggaacaatg gcatcgacat cataatcact taaacgatta
     9001 atatctaatt cttggtccac atacatgtct cttccatttt ggagataata gagatagagc
     9061 ttttcatttt gcaattgagt attttcaaca ggatgctctt taagaatctg acttcctaat
     9121 tctttgatac cttcttcgat tcgtttcata cgctctcgcg aatttttctg gcccttttga
     9181 gttgtctgat tttcacgtgc catttcaata acgatatttt ctggcttatg ccgccccatt
     9241 actttgacca attcatcaac aacttttaca gtctgtaaaa tacctttttt aatagcaggg
     9301 ctaccagcta aatttgcaat atgttcatgt aaactatcgc cttgtccaga cacttgtgct
     9361 ttttgaatgt cttctttaaa tgtcaaacta tcatcatgga tcagctgcat aaaattgcga
     9421 ttggcaaaac catctgattt caaaaaatct aatattgttt tgccagattg cttatcccta
     9481 ataccattaa tcaattttcg agacaaacgt ccccaaccag tataacggcg acgtttaagc
     9541 tgtttcatca ccttatcatc aaagaggtga gcatatgttt taagtctttc ctcaatcatc
     9601 tccctatctt caaataaggt caatgttaaa acaatatcct ctaagatatc ttcattttct
     9661 tcattatcca aaaaatcttt atctttaata atttttagca aatcatggta ggtacctaat
     9721 gaagcattaa atctatcttc aactcctgaa atttcaacac tatcaaaaca ttctattttt
     9781 ttgaaataat cttcttttaa ttgcttaacg gttacttttc gatttgtttt gaagagtaaa
     9841 tcaacaatgg ctttcttctg ttcacctgaa agaaatgctg gttttcgcat tccttcagta
     9901 acatatttga cctttgtcaa ttcgttataa accgtaaaat actcataaag caaactatgt
     9961 tttggtagta ctttttcatt tggaagattt ttatcaaagt ttgtcatgcg ttcaataaat
    10021 gattgagctg aagcaccttt atcgacaact tcttcaaaat tccatggggt aattgtttct
    10081 tcagacttcc gagtcatcca tgcaaaacga ctattgccac gcgccaatgg accaacataa
    10141 taagggattc gaaaagtcaa gattttttca atcttctcac gattgtcttt taaaaatgga
    10201 taaaagtctt cttgtcttct caaaatagca tgcagctcac ccaagtgaat ttgatgggga
    10261 atagagccgt tgtcaaaggt ccgttgcttg cgcagcaaat cttcacgatt tagtttcacc
    10321 aataattcct cagtaccatc cattttttct aaaattggtt tgataaattt ataaaattct
    10381 tcttggctag ctcccccatc aatataacct gcatatccgt tttttgattg atcaaaaaag
    10441 atttctttat acttttctgg aagttgttgt cgaactaaag cttttaaaag agtcaagtct
    10501 tgatgatgtt catcgtagcg tttaatcatt gaagctgata ggggagcctt agttatttca
    10561 gtatttactc ttaggatatc tgaaagtaaa atagcatctg ataaattctt agctgccaaa
    10621 aacaaatcag catattgatc tccaatttgc gccaataaat tatctaaatc atcatcgtaa
    10681 gtatcttttg aaagctgtaa tttagcatct tctgccaaat caaaatttga tttaaaatta
    10741 ggggtcaaac ccaatgacaa agcaatgaga ttcccaaata agccattttt cttctcaccg
    10801 gggagctgag caatgagatt ttctaatcgt cttgatttac tcaatcgtgc agaaagaatc
    10861 gctttagcat ctactccact tgcgttaata gggttttctt caaataattg attgtaggtt
    10921 tgtaccaact ggataaatag tttgtccaca tcactattat caggatttaa atctccctca
    10981 atcaaaaaat gaccacgaaa cttaatcata tgcgctaagg ccaaatagat taagcgcaaa
    11041 tccgctttat cagtagaatc taccaatttt tttcgcagat gatagatagt tggatatttc
    11101 tcatgataag caacttcatc tactatattt ccaaaaatag gatgacgttc atgcttcttg
    11161 tcttcttcca ccaaaaaaga ctcttcaagt cgatgaaaga aactatcatc tactttcgcc
    11221 atctcatttg aaaaaatctc ctgtagataa caaatacgat tcttccgacg tgtatacctt
    11281 ctacgagctg tccgtttgag acgagtcgct tccgctgtct ctccactgtc aaataaaaga
    11341 gcccctataa gatttttttt gatactgtgg cggtctgtat ttcccagaac cttgaacttt
    11401 ttagacggaa ccttatattc atcagtgatc accgcccatc cgacgctatt tgtgccgata
    11461 gctaagccta ttgagtattt cttatccata ctagtcctcc ttctcgctca attgttatcc
    11521 gctcacaatt gaatctaagt atcattgtta tccgctcaca agtcaacact ctttttgata
    11581 aattttacta gtctgcagac gtaaaaaaag cggcgtggtt agccgctttt ttaattgccg
    11641 gagcgcaata aaaaagcccc cggaaggtga tcttccgggg gctttctcat gcgttgcttg
    11701 tattgaaaat gggggccatg cctacctatc tgcctcccgg gaagcggcat gcatttacgt
    11761 t
//
