LOCUS       sequence-314828-       12005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mobile CRISPRi sfGFP 'test', empty sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12005)
  AUTHORS   Banta AB, Ward RD, Tran JS, Bacon EE, Peters JM
  TITLE     Programmable Gene Knockdown in Diverse Bacteria Using 
            Mobile-CRISPRi.
  JOURNAL   Curr Protoc Microbiol. 2020 Dec;59(1):e130. doi: 10.1002/cpmc.130.
  PUBMED    33332762
REFERENCE   2  (bases 1 to 12005)
  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: "Curr Protoc
            Microbiol."; date: "2020-12"; volume: "59(1):e130. doi"; pages: " 
            10.1002/cpmc.130"
COMMENT     Sequence Label: pJMP2822
FEATURES             Location/Qualifiers
     source          1..12005
                     /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    1446..1462
                     /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)."
     CDS             join(1837..1839,1840..2553)
                     /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             1837..2550
                     /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(1867..1895)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2490..2510
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    complement(2695..2742)
                     /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(2819..2824,2825..2841,2842..2847)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2958..3752
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3012..3031)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3622..3641
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    complement(3804..3851)
                     /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(3902..3988)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(4091..4185)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(4213..4229)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4214..4230)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4644..4663)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4763..4785
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4823..4841)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4909..5013
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5014..5082,5083..5874)
                     /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(5232..5251)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6175..6563
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            6832..6940
                     /note="incP origin of transfer"
     mobile_element  7044..7209
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     CDS             complement(7364..7453)
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             complement(join(7463..9046,9047..9049,9050..11536,
                     11537..11539,11540..11566))
                     /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    11608..11624
                     /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 gatggtacgc 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 atagctcatg 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 aaagaattcc taaagatcta taaatgtgag cggataacat
     1441 tgacattgtg agcggataac aagatacttc tagttgagac caactttggt ctccaccata
     1501 gcggtcggtc tctgtttaag agctatgctg gaaacagcat agcaagttta aataaggcta
     1561 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttttga attcatgtgg
     1621 ctgaccgttc tgttgtctct cgctcttccg agtagacgaa caataaggcc tccctaacgg
     1681 ggggcctttt ttattgataa caaaagtcag tgcttccgct atttccaaaa taccgggcta
     1741 atacggttta aacgaaaatt tatcaaaaag agtattgact taaagtctaa cctataggat
     1801 acttacagcc agatctgagc gagaaggagg taaagtatga gcaaaggaga agaacttttc
     1861 actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct
     1921 gtccgtggag agggtgaagg tgatgctaca aacggaaaac tcacccttaa atttatttgc
     1981 actactggaa aactacctgt tccgtggcca acacttgtca ctactctgac ctatggtgtt
     2041 caatgctttt cccgttatcc ggatcacatg aaacggcatg actttttcaa gagtgccatg
     2101 cccgaaggtt atgtacagga acgcactata tctttcaaag atgacgggac ctacaagacg
     2161 cgtgctgaag tcaagtttga aggtgatacc cttgttaatc gtatcgagtt aaagggtatt
     2221 gattttaaag aagatggaaa cattcttgga cacaaactgg agtacaactt taactcacac
     2281 aatgtataca tcacggcaga caaacaaaag aatggaatca aagctaactt caaaattcgc
     2341 cacaacgttg aagatggttc cgttcaacta gcagaccatt atcaacaaaa tactccaatt
     2401 ggcgatggcc ctgtcctttt accagacaac cattacctgt cgacacaatc tgtcctttcg
     2461 aaagatccca acgaaaagcg tgaccacatg gtccttcttg agtttgtaac tgctgctggg
     2521 attacacatg gcatggatga gctctacaaa taataagttt aaacgacctc ctggatttgc
     2581 tcagacagcc ttttcgtcat tcgtttcagc caaaaaactt aagaccgccg gtcttgtcca
     2641 ctaccttgca gtaatgcggt ggacaggatc ggcggttttc ttttctcttc tcaagaagtt
     2701 cctatacttt ctagagaata ggaacttcgg aataggaact tcctcctgaa cggccataag
     2761 aacgaaggct gtctgttgaa ctctcgagcc gtggaaacgg atgaaggcac gaacccagtt
     2821 gacataagcc tgttcggttc gtaaactgta atgcaagtag cgtatgcgct caaaactgga
     2881 tggctttctt gccgccaagg atctgatggc gcaggggatc aagatctgat caagagacag
     2941 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     3001 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     3061 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     3121 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     3181 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     3241 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     3301 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3361 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3421 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3481 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3541 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3601 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3661 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3721 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgactcg
     3781 agcatcagct tcaaaagcgc tctgaagttc ctatactttc tagagaatag gaacttcgga
     3841 ataggtactt caagatcccc aattcgagct catcgtccgg gccgcaagct cctagcggcg
     3901 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
     3961 ctttcgactg agcctttcgt tttatttgat gcctcaagct agagagtcat taccccaggc
     4021 gtttaagggc accaataact gccttaaaaa aattacgccc cgccctgcca ctcatcgcag
     4081 tctagcttgg attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat
     4141 ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagc tcagctaatt
     4201 aagctagctt atcgataccg tcgacctcga accccacgcc cctctttaat acgacgggca
     4261 atttgcactt cagaaaatga agagtttgct ttagccataa caaaagtcca gtatgctttt
     4321 tcacagcata actggactga tttcagttta caactattct gtctagttta agactttatt
     4381 gtcatagttt agatctattt tgttcagttt aagactttat tgtccgccca cacctgctta
     4441 cgcagggcat ccatttatta ctcaaccgta accgattttg ccaggttacg cggctggtcc
     4501 tctagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     4561 ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4621 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     4681 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4741 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcgtca
     4801 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4861 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4921 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4981 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5041 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5101 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5161 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5221 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5281 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5341 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5401 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5461 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5521 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5581 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5641 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5701 gctgataaat ctggagccgg tgagcgtggg tcccgcggta tcattgcagc actggggcca
     5761 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5821 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5881 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5941 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6001 ttccactgag cgtcagaccc cgtagaaaag atcaaaggca gttattggtg cctcactgat
     6061 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6121 tcatttttaa tttttgcggc cgcaagatcc ggccacgatg cgtccggcgt agaggatctg
     6181 aagatcagca gttcaacctg ttgatagtac gtactaagct ctcatgtttc acgtactaag
     6241 ctctcatgtt taacgtacta agctctcatg tttaacgaac taaaccctca tggctaacgt
     6301 actaagctct catggctaac gtactaagct ctcatgtttc acgtactaag ctctcatgtt
     6361 tgaacaataa aattaatata aatcagcaac ttaaatagcc tctaaggttt taagttttat
     6421 aagaaaaaaa agaatatata aggcttttaa agcttttaag gtttaacggt tgtggacaac
     6481 aagccaggga tgtaacgcac tgagaagccc ttagagcctc tcaaagcaat tttgagtgac
     6541 acaggaacac ttaacggctg acatgggaat tagccatggc atcacagtat cgtgatgaca
     6601 gaggcaggga gtgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6661 ttctttcctg cgttatcagg ggattcctta aggtatactt tccgctgcat aaccctgctt
     6721 cggggtcatt atagcgattt tttcggtata tccatccttt ttcgcacgat atacaggatt
     6781 ttgccaaagg gttcgtgtag actttccttg gtgtatccaa cggcgtcagc cgggcaggat
     6841 aggtgaagta ggcccacccg cgagcgggtg ttccttcttc actgtccctt attcgcacct
     6901 ggcggtgctc aacgggaatc ctgctctgcg aggctggccg ataagctctg ataccgctcg
     6961 ccgcagccga acgaccgagc gcagcgggtc agtgagcgag gaagcggaag agcccaaaca
     7021 taacaggaag aaaaatgccc cgctgtgggc ggacaaaata gttgggaact gggaggggtg
     7081 gaaatggagt ttttaaggat tatttaggga agagtgacaa aatagatggg aactgggtgt
     7141 agcgtcgtaa gctaatacga aaattaaaaa tgacaaaata gtttggaact agatttcact
     7201 tatctggttc ttgaggcggt taaaagagcc gtactcttct ccgatgtcga ctaggccatg
     7261 atgctcattc tgtgggacca aaacgaaaaa acaccctttc gggtgtcttt tctggaattt
     7321 ggtaccgagg ctgcaagtcc cattaaggag gcgcgcctta ttacagatct tcctcagaga
     7381 tgagcttctg ttccagatct tcttctgaaa tcaacttttg ttccagatct tcttcagaga
     7441 tgagtttctg ctccgcggcc gcgtcacctc ctagctgact caaatcaatg cgtgtttcat
     7501 aaagaccagt gatggattga tggataagag tggcatctaa aacttctttt gtagacgtat
     7561 atcgtttacg atcaattgtt gtatcaaaat atttaaaagc agcgggagct ccaagattcg
     7621 tcaacgtaaa taaatgaata atattttctg cttgttcacg tattggtttg tctctatgtt
     7681 tgttatatgc actaagaact ttatctaaat tggcatctgc taaaataaca cgcttagaaa
     7741 attcactgat ttgctcaata atctcatcta aataatgctt atgctgctcc acaaacaatt
     7801 gtttttgttc gttatcttct ggactaccct tcaacttttc ataatgacta gctaaatata
     7861 aaaaattcac atatttgctt ggcagagcca gctcatttcc tttttgtaat tctccggcac
     7921 tagccagcat ccgtttacga ccgttttcta actcaaaaag actatattta ggtagtttaa
     7981 tgattaagtc ttttttaact tccttatatc ctttagcttc taaaaagtca atcggatttt
     8041 tttcaaagga acttctttcc ataattgtga tccctagtaa ctctttaacg gattttaact
     8101 tcttcgattt ccctttttcc accttagcaa ccactaggac tgaataagct accgttggac
     8161 tatcaaaacc accatatttt tttggatccc agtctttttt acgagcaata agcttgtccg
     8221 aatttctttt tggtaaaatt gactccttgg agaatccgcc tgtctgtact tctgttttct
     8281 tgacaatatt gacttggggc atggacaata ctttgcgcac tgtggcaaaa tctcgccctt
     8341 tatcccagac aatttctcca gtttccccat tagtttcgat tagagggcgt ttgcgaatct
     8401 ctccatttgc aagtgtaatt tctgttttga agaagttcat gatattagag taaaagaaat
     8461 attttgcggt tgctttgcct atttcttgct cagacttagc aatcatttta cgaacatcat
     8521 aaactttata atcaccatag acaaactccg attcaagttt tggatatttc ttaatcaaag
     8581 cagttccaac gacggcattt agatacgcat catgggcatg atggtaattg ttaatctcac
     8641 gtactttata gaattggaaa tcttttcgga agtcagaaac taatttagat tttaaggtaa
     8701 tcactttaac ctctcgaata agtttatcat tttcatcgta tttagtattc atgcgactat
     8761 ccaaaatttg tgccacatgc ttagtgattt ggcgagtttc aaccaattgg cgtttgataa
     8821 aaccagcttt atcaagttca ctcaaacctc cacgttcagc tttcgttaaa ttatcaaact
     8881 tacgttgagt gattaacttg gcgtttagaa gttgtctcca atagtttttc atctttttga
     8941 ctacttcttc acttggaacg ttatccgatt taccacgatt tttatcagaa cgcgttaaga
     9001 ccttattgtc tattgaatcg tctttaagga aactttgtgg aacaatggca tcgacatcat
     9061 aatcacttaa acgattaata tctaattctt ggtccacata catgtctctt ccattttgga
     9121 gataatagag atagagcttt tcattttgca attgagtatt ttcaacagga tgctctttaa
     9181 gaatctgact tcctaattct ttgatacctt cttcgattcg tttcatacgc tctcgcgaat
     9241 ttttctggcc cttttgagtt gtctgatttt cacgtgccat ttcaataacg atattttctg
     9301 gcttatgccg ccccattact ttgaccaatt catcaacaac ttttacagtc tgtaaaatac
     9361 cttttttaat agcagggcta ccagctaaat ttgcaatatg ttcatgtaaa ctatcgcctt
     9421 gtccagacac ttgtgctttt tgaatatctt ctttaaatgt caaactatca tcatggatca
     9481 gctgcataaa attgcgattg gcaaaaccat ctgatttcaa aaaatctaat attgttttgc
     9541 cagattgctt atccctaata ccattaatca attttcgaga caaacgtccc caaccagtat
     9601 aacggcgacg tttaagctgt ttcatcacct tatcatcaaa gaggtgagca tatgttttaa
     9661 gtctttcctc aatcatctcc ctatcttcaa ataaggtcaa tgttaaaaca atatcctcta
     9721 agatatcttc attttcttca ttatccaaaa aatctttatc tttaataatt tttagcaaat
     9781 catggtaggt acctaatgaa gcattaaatc tatcttcaac tcctgaaatt tcaacactat
     9841 caaaacattc tatttttttg aaataatctt cttttaattg cttaacggtt acttttcgat
     9901 ttgttttgaa gagtaaatca acaatggctt tcttctgttc acctgaaaga aatgctggtt
     9961 ttcgcattcc ttcagtaaca tatttgacct ttgtcaattc gttataaacc gtaaaatact
    10021 cataaagcaa actatgtttt ggtagtactt tttcatttgg aagattttta tcaaagtttg
    10081 tcatgcgttc aataaatgat tgagctgaag cacctttatc gacaacttct tcaaaattcc
    10141 atggggtaat tgtttcttca gacttccgag tcatccatgc aaaacgacta ttgccacgcg
    10201 ccaatggacc aacataataa gggattcgaa aagtcaagat tttttcaatc ttctcacgat
    10261 tgtcttttaa aaatggataa aaatcttctt gtcgtctcaa aatagcatgc agctcaccca
    10321 agtgaatttg atggggaata gagccgttgt caaaggtccg ttgcttgcgc agcaaatctt
    10381 cacgatttag tttcaccaat aattcctcag taccatccat tttttctaaa attggtttga
    10441 taaatttata aaattcttct tggctagctc ccccatcaat ataacctgca tatccgtttt
    10501 ttgattgatc aaaaaagatt tctttatact tttctggaag ttgttgtcga actaaagctt
    10561 ttaaaagagt caagtcttga tgatgttcat cgtagcgttt aatcattgaa gctgataggg
    10621 gagccttagt tatttcagta tttactctta ggatatctga aagtaaaata gcatctgata
    10681 aattcttagc tgccaaaaac aaatcagcat attgatctcc aatttgcgcc aataaattat
    10741 ctaaatcatc atcgtaagta tcttttgaaa gctgtaattt agcatcttct gccaaatcaa
    10801 aatttgattt aaaattaggg gtcaaaccca atgacaaagc aatgagattc ccaaataagc
    10861 catttttctt ctcaccgggg agctgagcaa tgagattttc taatcgtctt gatttactca
    10921 atcgtgcaga aagaatcgct ttagcatcta ctccacttgc gttaataggg ttttcttcaa
    10981 ataattgatt gtaggtttgt accaactgga taaatagttt gtccacatca ctattatcag
    11041 gatttaaatc tccctcaatc aaaaaatgac cacgaaactt aatcatatgc gctaaggcca
    11101 aatagattaa gcgcaaatcc gctttatcag tagaatctac caattttttt cgcagatgat
    11161 agatagttgg atatttctca tgataagcaa cttcatctac tatatttcca aaaataggat
    11221 gacgttcatg cttcttatct tcttccacca aaaaagactc ttcaagtcga tgaaagaaac
    11281 tatcatctac tttcgccatc tcatttgaaa aaatctcctg tagataacaa atacgattct
    11341 tccgacgtgt ataccttcta cgagctgtcc gtttgagacg agtcgcttcc gctgtctctc
    11401 cactgtcaaa taaaagagcc cctataagat tttttttgat actgtggcgg tctgtatttc
    11461 ccagaacctt gaacttttta gacggaacct tatattcatc agtgatcacc gcccatccga
    11521 cgctatttgt gccgatagct aagcctattg agtatttctt atccatacta gttttcctgt
    11581 gtgaaacctg ctgatgtgct cagtatcttg ttatccgctc acaatgtcaa tgttatccgc
    11641 tcacatttat aatattttat ctgattaata agatgatctt cttgagatcg ttttggtctg
    11701 cgcgtaatct cttgctctga aaacgaaaaa accgccttgc agggcggttt ttcgaaggtt
    11761 ctctgagcta ccaactcttt gaaccgaggt aactggcttg gtgataagct gtcaaaccag
    11821 atcaattcgc actagtctgc agacgtaaaa aaagcggcgt ggttagccgc ttttttaatt
    11881 gccggagcgc aataaaaaag cccccggaag gtgatcttcc gggggctttc tcatgcgttg
    11941 cttgtattga aaatgggggc catgcctacc tatctgcctc ccgggaagcg gcatgcattt
    12001 acgtt
//
