LOCUS       sequence-314830-       11809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mobile CRISPRi sfGFP 'test', gmc6 sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11809)
  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 11809)
  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: pJMP2832
FEATURES             Location/Qualifiers
     source          1..11809
                     /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(1818..1820,1821..2534)
                     /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             1818..2531
                     /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(1848..1876)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2471..2491
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    complement(2676..2723)
                     /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(2800..2805,2806..2822,2823..2828)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2911..3570
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2977..2996)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    complement(3608..3655)
                     /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(3706..3792)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(3895..3989)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(4017..4033)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4018..4034)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4448..4467)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4567..4589
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4627..4645)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4713..4817
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4818..4886,4887..5678)
                     /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(5036..5055)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5979..6367
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            6636..6744
                     /note="incP origin of transfer"
     mobile_element  6848..7013
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     CDS             complement(7168..7257)
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             complement(join(7267..8850,8851..8853,8854..11340,
                     11341..11343,11344..11370))
                     /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    11412..11428
                     /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 tagtcatcta attcaacaag aattgtttaa
     1501 gagctatgct ggaaacagca tagcaagttt aaataaggct agtccgttat caacttgaaa
     1561 aagtggcacc gagtcggtgc tttttttttg aattcatgtg gctgaccgtt ctgttgtctc
     1621 tcgctcttcc gagtagacga acaataaggc ctccctaacg gggggccttt tttattgata
     1681 acaaaagtca gtgcttccgc tatttccaaa ataccgggct aatacggttt aaacgaaaat
     1741 ttatcaaaaa gagtattgac ttaaagtcta acctatagga tacttacagc cagatctgag
     1801 cgagaaggag gtaaagtatg agcaaaggag aagaactttt cactggagtt gtcccaattc
     1861 ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtccgtgga gagggtgaag
     1921 gtgatgctac aaacggaaaa ctcaccctta aatttatttg cactactgga aaactacctg
     1981 ttccgtggcc aacacttgtc actactctga cctatggtgt tcaatgcttt tcccgttatc
     2041 cggatcacat gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg
     2101 aacgcactat atctttcaaa gatgacggga cctacaagac gcgtgctgaa gtcaagtttg
     2161 aaggtgatac ccttgttaat cgtatcgagt taaagggtat tgattttaaa gaagatggaa
     2221 acattcttgg acacaaactg gagtacaact ttaactcaca caatgtatac atcacggcag
     2281 acaaacaaaa gaatggaatc aaagctaact tcaaaattcg ccacaacgtt gaagatggtt
     2341 ccgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt
     2401 taccagacaa ccattacctg tcgacacaat ctgtcctttc gaaagatccc aacgaaaagc
     2461 gtgaccacat ggtccttctt gagtttgtaa ctgctgctgg gattacacat ggcatggatg
     2521 agctctacaa ataataagtt taaacgacct cctggatttg ctcagacagc cttttcgtca
     2581 ttcgtttcag ccaaaaaact taagaccgcc ggtcttgtcc actaccttgc agtaatgcgg
     2641 tggacaggat cggcggtttt cttttctctt ctcaagaagt tcctatactt tctagagaat
     2701 aggaacttcg gaataggaac ttcctcctga acggccataa gaacgaaggc tgtctgttga
     2761 actctcgagc cgtggaaacg gatgaaggca cgaacccagt tgacataagc ctgttcggtt
     2821 cgtaaactgt aatgcaagta gcgtatgcgc tcatcactac cgggcgtatt ttttgagtta
     2881 tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc
     2941 gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa
     3001 tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa
     3061 aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat
     3121 ccggagttcc gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct
     3181 tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac
     3241 gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac
     3301 ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg
     3361 gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt
     3421 ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag
     3481 gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag
     3541 tactgcgatg agtggcaggg cggggcgtaa tcgactggaa ctcgagcatc agcttcaaaa
     3601 gcgctctgaa gttcctatac tttctagaga ataggaactt cggaataggt acttcaagat
     3661 ccccaattcg agctcatcgt ccgggccgca agctcctagc ggcggatttg tcctactcag
     3721 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtctttcg actgagcctt
     3781 tcgttttatt tgatgcctca agctagagag tcattacccc aggcgtttaa gggcaccaat
     3841 aactgcctta aaaaaattac gccccgccct gccactcatc gcagtctagc ttggattctc
     3901 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
     3961 ggtcattact ggatctatca acaggagtcc aagctcagct aattaagcta gcttatcgat
     4021 accgtcgacc tcgaacccca cgcccctctt taatacgacg ggcaatttgc acttcagaaa
     4081 atgaagagtt tgctttagcc ataacaaaag tccagtatgc tttttcacag cataactgga
     4141 ctgatttcag tttacaacta ttctgtctag tttaagactt tattgtcata gtttagatct
     4201 attttgttca gtttaagact ttattgtccg cccacacctg cttacgcagg gcatccattt
     4261 attactcaac cgtaaccgat tttgccaggt tacgcggctg gtcctctagc tggcgtaata
     4321 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     4381 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
     4441 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     4501 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     4561 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc gtcaccgaaa cgcgcgagac
     4621 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     4681 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4741 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4801 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4861 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4921 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4981 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5041 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5101 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5161 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5221 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5281 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5341 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5401 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5461 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5521 ccggtgagcg tgggtcccgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5581 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5641 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5701 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5761 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5821 accccgtaga aaagatcaaa ggcagttatt ggtgcctcac tgattaagca ttggtaactg
     5881 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttttg
     5941 cggccgcaag atccggccac gatgcgtccg gcgtagagga tctgaagatc agcagttcaa
     6001 cctgttgata gtacgtacta agctctcatg tttcacgtac taagctctca tgtttaacgt
     6061 actaagctct catgtttaac gaactaaacc ctcatggcta acgtactaag ctctcatggc
     6121 taacgtacta agctctcatg tttcacgtac taagctctca tgtttgaaca ataaaattaa
     6181 tataaatcag caacttaaat agcctctaag gttttaagtt ttataagaaa aaaaagaata
     6241 tataaggctt ttaaagcttt taaggtttaa cggttgtgga caacaagcca gggatgtaac
     6301 gcactgagaa gcccttagag cctctcaaag caattttgag tgacacagga acacttaacg
     6361 gctgacatgg gaattagcca tggcatcaca gtatcgtgat gacagaggca gggagtgcgg
     6421 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6481 caggggattc cttaaggtat actttccgct gcataaccct gcttcggggt cattatagcg
     6541 attttttcgg tatatccatc ctttttcgca cgatatacag gattttgcca aagggttcgt
     6601 gtagactttc cttggtgtat ccaacggcgt cagccgggca ggataggtga agtaggccca
     6661 cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc acctggcggt gctcaacggg
     6721 aatcctgctc tgcgaggctg gccgataagc tctgataccg ctcgccgcag ccgaacgacc
     6781 gagcgcagcg ggtcagtgag cgaggaagcg gaagagccca aacataacag gaagaaaaat
     6841 gccccgctgt gggcggacaa aatagttggg aactgggagg ggtggaaatg gagtttttaa
     6901 ggattattta gggaagagtg acaaaataga tgggaactgg gtgtagcgtc gtaagctaat
     6961 acgaaaatta aaaatgacaa aatagtttgg aactagattt cacttatctg gttcttgagg
     7021 cggttaaaag agccgtactc ttctccgatg tcgactaggc catgatgctc attctgtggg
     7081 accaaaacga aaaaacaccc tttcgggtgt cttttctgga atttggtacc gaggctgcaa
     7141 gtcccattaa ggaggcgcgc cttattacag atcttcctca gagatgagct tctgttccag
     7201 atcttcttct gaaatcaact tttgttccag atcttcttca gagatgagtt tctgctccgc
     7261 ggccgcgtca cctcctagct gactcaaatc aatgcgtgtt tcataaagac cagtgatgga
     7321 ttgatggata agagtggcat ctaaaacttc ttttgtagac gtatatcgtt tacgatcaat
     7381 tgttgtatca aaatatttaa aagcagcggg agctccaaga ttcgtcaacg taaataaatg
     7441 aataatattt tctgcttgtt cacgtattgg tttgtctcta tgtttgttat atgcactaag
     7501 aactttatct aaattggcat ctgctaaaat aacacgctta gaaaattcac tgatttgctc
     7561 aataatctca tctaaataat gcttatgctg ctccacaaac aattgttttt gttcgttatc
     7621 ttctggacta cccttcaact tttcataatg actagctaaa tataaaaaat tcacatattt
     7681 gcttggcaga gccagctcat ttcctttttg taattctccg gcactagcca gcatccgttt
     7741 acgaccgttt tctaactcaa aaagactata tttaggtagt ttaatgatta agtctttttt
     7801 aacttcctta tatcctttag cttctaaaaa gtcaatcgga tttttttcaa aggaacttct
     7861 ttccataatt gtgatcccta gtaactcttt aacggatttt aacttcttcg atttcccttt
     7921 ttccacctta gcaaccacta ggactgaata agctaccgtt ggactatcaa aaccaccata
     7981 tttttttgga tcccagtctt ttttacgagc aataagcttg tccgaatttc tttttggtaa
     8041 aattgactcc ttggagaatc cgcctgtctg tacttctgtt ttcttgacaa tattgacttg
     8101 gggcatggac aatactttgc gcactgtggc aaaatctcgc cctttatccc agacaatttc
     8161 tccagtttcc ccattagttt cgattagagg gcgtttgcga atctctccat ttgcaagtgt
     8221 aatttctgtt ttgaagaagt tcatgatatt agagtaaaag aaatattttg cggttgcttt
     8281 gcctatttct tgctcagact tagcaatcat tttacgaaca tcataaactt tataatcacc
     8341 atagacaaac tccgattcaa gttttggata tttcttaatc aaagcagttc caacgacggc
     8401 atttagatac gcatcatggg catgatggta attgttaatc tcacgtactt tatagaattg
     8461 gaaatctttt cggaagtcag aaactaattt agattttaag gtaatcactt taacctctcg
     8521 aataagttta tcattttcat cgtatttagt attcatgcga ctatccaaaa tttgtgccac
     8581 atgcttagtg atttggcgag tttcaaccaa ttggcgtttg ataaaaccag ctttatcaag
     8641 ttcactcaaa cctccacgtt cagctttcgt taaattatca aacttacgtt gagtgattaa
     8701 cttggcgttt agaagttgtc tccaatagtt tttcatcttt ttgactactt cttcacttgg
     8761 aacgttatcc gatttaccac gatttttatc agaacgcgtt aagaccttat tgtctattga
     8821 atcgtcttta aggaaacttt gtggaacaat ggcatcgaca tcataatcac ttaaacgatt
     8881 aatatctaat tcttggtcca catacatgtc tcttccattt tggagataat agagatagag
     8941 cttttcattt tgcaattgag tattttcaac aggatgctct ttaagaatct gacttcctaa
     9001 ttctttgata ccttcttcga ttcgtttcat acgctctcgc gaatttttct ggcccttttg
     9061 agttgtctga ttttcacgtg ccatttcaat aacgatattt tctggcttat gccgccccat
     9121 tactttgacc aattcatcaa caacttttac agtctgtaaa ataccttttt taatagcagg
     9181 gctaccagct aaatttgcaa tatgttcatg taaactatcg ccttgtccag acacttgtgc
     9241 tttttgaata tcttctttaa atgtcaaact atcatcatgg atcagctgca taaaattgcg
     9301 attggcaaaa ccatctgatt tcaaaaaatc taatattgtt ttgccagatt gcttatccct
     9361 aataccatta atcaattttc gagacaaacg tccccaacca gtataacggc gacgtttaag
     9421 ctgtttcatc accttatcat caaagaggtg agcatatgtt ttaagtcttt cctcaatcat
     9481 ctccctatct tcaaataagg tcaatgttaa aacaatatcc tctaagatat cttcattttc
     9541 ttcattatcc aaaaaatctt tatctttaat aatttttagc aaatcatggt aggtacctaa
     9601 tgaagcatta aatctatctt caactcctga aatttcaaca ctatcaaaac attctatttt
     9661 tttgaaataa tcttctttta attgcttaac ggttactttt cgatttgttt tgaagagtaa
     9721 atcaacaatg gctttcttct gttcacctga aagaaatgct ggttttcgca ttccttcagt
     9781 aacatatttg acctttgtca attcgttata aaccgtaaaa tactcataaa gcaaactatg
     9841 ttttggtagt actttttcat ttggaagatt tttatcaaag tttgtcatgc gttcaataaa
     9901 tgattgagct gaagcacctt tatcgacaac ttcttcaaaa ttccatgggg taattgtttc
     9961 ttcagacttc cgagtcatcc atgcaaaacg actattgcca cgcgccaatg gaccaacata
    10021 ataagggatt cgaaaagtca agattttttc aatcttctca cgattgtctt ttaaaaatgg
    10081 ataaaaatct tcttgtcgtc tcaaaatagc atgcagctca cccaagtgaa tttgatgggg
    10141 aatagagccg ttgtcaaagg tccgttgctt gcgcagcaaa tcttcacgat ttagtttcac
    10201 caataattcc tcagtaccat ccattttttc taaaattggt ttgataaatt tataaaattc
    10261 ttcttggcta gctcccccat caatataacc tgcatatccg ttttttgatt gatcaaaaaa
    10321 gatttcttta tacttttctg gaagttgttg tcgaactaaa gcttttaaaa gagtcaagtc
    10381 ttgatgatgt tcatcgtagc gtttaatcat tgaagctgat aggggagcct tagttatttc
    10441 agtatttact cttaggatat ctgaaagtaa aatagcatct gataaattct tagctgccaa
    10501 aaacaaatca gcatattgat ctccaatttg cgccaataaa ttatctaaat catcatcgta
    10561 agtatctttt gaaagctgta atttagcatc ttctgccaaa tcaaaatttg atttaaaatt
    10621 aggggtcaaa cccaatgaca aagcaatgag attcccaaat aagccatttt tcttctcacc
    10681 ggggagctga gcaatgagat tttctaatcg tcttgattta ctcaatcgtg cagaaagaat
    10741 cgctttagca tctactccac ttgcgttaat agggttttct tcaaataatt gattgtaggt
    10801 ttgtaccaac tggataaata gtttgtccac atcactatta tcaggattta aatctccctc
    10861 aatcaaaaaa tgaccacgaa acttaatcat atgcgctaag gccaaataga ttaagcgcaa
    10921 atccgcttta tcagtagaat ctaccaattt ttttcgcaga tgatagatag ttggatattt
    10981 ctcatgataa gcaacttcat ctactatatt tccaaaaata ggatgacgtt catgcttctt
    11041 atcttcttcc accaaaaaag actcttcaag tcgatgaaag aaactatcat ctactttcgc
    11101 catctcattt gaaaaaatct cctgtagata acaaatacga ttcttccgac gtgtatacct
    11161 tctacgagct gtccgtttga gacgagtcgc ttccgctgtc tctccactgt caaataaaag
    11221 agcccctata agattttttt tgatactgtg gcggtctgta tttcccagaa ccttgaactt
    11281 tttagacgga accttatatt catcagtgat caccgcccat ccgacgctat ttgtgccgat
    11341 agctaagcct attgagtatt tcttatccat actagttttc ctgtgtgaaa cctgctgatg
    11401 tgctcagtat cttgttatcc gctcacaatg tcaatgttat ccgctcacat ttataatatt
    11461 ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct
    11521 ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga gctaccaact
    11581 ctttgaaccg aggtaactgg cttggtgata agctgtcaaa ccagatcaat tcgcactagt
    11641 ctgcagacgt aaaaaaagcg gcgtggttag ccgctttttt aattgccgga gcgcaataaa
    11701 aaagcccccg gaaggtgatc ttccgggggc tttctcatgc gttgcttgta ttgaaaatgg
    11761 gggccatgcc tacctatctg cctcccggga agcggcatgc atttacgtt
//
