LOCUS       sequence-314799-       11764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Z. mobilis CRISPRi, promoter A, empty guide, sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11764)
  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 11764)
  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: pJMP2046
FEATURES             Location/Qualifiers
     source          1..11764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..24,25..31)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             join(400..402,403..1116)
                     /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             400..1113
                     /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(430..458)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1053..1073
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    complement(1258..1305)
                     /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(1382..1387,1388..1404,1405..1410)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     primer_bind     complement(1559..1578)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    complement(2242..2289)
                     /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(2340..2426)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(2529..2623)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(2651..2667)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2652..2668)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3082..3101)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3201..3223
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3261..3279)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3347..3451
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3452..3520,3521..4312)
                     /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(3670..3689)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4613..5001
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            5270..5378
                     /note="incP origin of transfer"
     mobile_element  5537..5702
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     CDS             complement(5857..5946)
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             complement(join(5956..7539,7540..7542,7543..10029,
                     10030..10032,10033..10059))
                     /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    10082..10098
                     /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    10115..10131
                     /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        10332..10409
                     /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             10410..11492
                     /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(10429..10448)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tttacacttt atgcttccgg ctcgtataat gtctagttga gaccaacttt ggtctccacc
       61 atagcggtcg gtctctgttt aagagctatg ctggaaacag catagcaagt ttaaataagg
      121 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttt tgaattcatg
      181 tggctgaccg ttctgttgtc tctcgctctt ccgagtagac gaacaataag gcctccctaa
      241 cggggggcct tttttattga taacaaaagt cagtgcttcc gctatttcca aaataccggg
      301 ctaatacggt ttaaacgaaa atttatcaaa aagagtattg acttaaagtc taacctatag
      361 gatacttaca gccagatctg agcgagaagg aggtaaagta tgagcaaagg agaagaactt
      421 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
      481 tctgtccgtg gagagggtga aggtgatgct acaaacggaa aactcaccct taaatttatt
      541 tgcactactg gaaaactacc tgttccgtgg ccaacacttg tcactactct gacctatggt
      601 gttcaatgct tttcccgtta tccggatcac atgaaacggc atgacttttt caagagtgcc
      661 atgcccgaag gttatgtaca ggaacgcact atatctttca aagatgacgg gacctacaag
      721 acgcgtgctg aagtcaagtt tgaaggtgat acccttgtta atcgtatcga gttaaagggt
      781 attgatttta aagaagatgg aaacattctt ggacacaaac tggagtacaa ctttaactca
      841 cacaatgtat acatcacggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
      901 cgccacaacg ttgaagatgg ttccgttcaa ctagcagacc attatcaaca aaatactcca
      961 attggcgatg gccctgtcct tttaccagac aaccattacc tgtcgacaca atctgtcctt
     1021 tcgaaagatc ccaacgaaaa gcgtgaccac atggtccttc ttgagtttgt aactgctgct
     1081 gggattacac atggcatgga tgagctctac aaataataag tttaaacgac ctcctggatt
     1141 tgctcagaca gccttttcgt cattcgtttc agccaaaaaa cttaagaccg ccggtcttgt
     1201 ccactacctt gcagtaatgc ggtggacagg atcggcggtt ttcttttctc ttctcaagaa
     1261 gttcctatac tttctagaga ataggaactt cggaatagga acttcctcct gaacggccat
     1321 aagaacgaag gctgtctgtt gaactctcga gccgtggaaa cggatgaagg cacgaaccca
     1381 gttgacataa gcctgttcgg ttcgtaaact gtaatgcaag tagcgtatgc gctcatcact
     1441 accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatggagaa
     1501 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
     1561 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
     1621 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
     1681 tgcccgcctg atgaatgctc atccggagtt ccgtatggca atgaaagacg gtgagctggt
     1741 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
     1801 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga
     1861 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
     1921 tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat
     1981 ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag gcgacaaggt
     2041 gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag
     2101 aatgcttaat gaattacaac agtttttatg catgcgccca atacgcaaac cgcctctccc
     2161 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaactcgag
     2221 catcagcttc aaaagcgctc tgaagttcct atactttcta gagaatagga acttcggaat
     2281 aggtacttca agatccccaa ttcgagctca tcgtccgggc cgcaagctcc tagcggcgga
     2341 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     2401 ttcgactgag cctttcgttt tatttgatgc ctcaagctag agagtcatta ccccaggcgt
     2461 ttaagggcac caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcagtc
     2521 tagcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
     2581 agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagctc agctaattaa
     2641 gctagcttat cgataccgtc gacctcgaac cccacgcccc tctttaatac gacgggcaat
     2701 ttgcacttca gaaaatgaag agtttgcttt agccataaca aaagtccagt atgctttttc
     2761 acagcataac tggactgatt tcagtttaca actattctgt ctagtttaag actttattgt
     2821 catagtttag atctattttg ttcagtttaa gactttattg tccgcccaca cctgcttacg
     2881 cagggcatcc atttattact caaccgtaac cgattttgcc aggttacgcg gctggtcctc
     2941 tagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     3001 gaatggcgaa tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3061 ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     3121 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     3181 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcgtcacc
     3241 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     3301 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3361 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3421 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     3481 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     3541 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     3601 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     3661 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     3721 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     3781 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     3841 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     3901 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     3961 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     4021 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     4081 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     4141 tgataaatct ggagccggtg agcgtgggtc ccgcggtatc attgcagcac tggggccaga
     4201 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     4261 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     4321 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     4381 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     4441 ccactgagcg tcagaccccg tagaaaagat caaaggcagt tattggtgcc tcactgatta
     4501 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4561 atttttaatt tttgcggccg caagatccgg ccacgatgcg tccggcgtag aggatctgaa
     4621 gatcagcagt tcaacctgtt gatagtacgt actaagctct catgtttcac gtactaagct
     4681 ctcatgttta acgtactaag ctctcatgtt taacgaacta aaccctcatg gctaacgtac
     4741 taagctctca tggctaacgt actaagctct catgtttcac gtactaagct ctcatgtttg
     4801 aacaataaaa ttaatataaa tcagcaactt aaatagcctc taaggtttta agttttataa
     4861 gaaaaaaaag aatatataag gcttttaaag cttttaaggt ttaacggttg tggacaacaa
     4921 gccagggatg taacgcactg agaagccctt agagcctctc aaagcaattt tgagtgacac
     4981 aggaacactt aacggctgac atgggaatta gccatggcat cacagtatcg tgatgacaga
     5041 ggcagggagt gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5101 ctttcctgcg ttatcagggg attccttaag gtatactttc cgctgcataa ccctgcttcg
     5161 gggtcattat agcgattttt tcggtatatc catccttttt cgcacgatat acaggatttt
     5221 gccaaagggt tcgtgtagac tttccttggt gtatccaacg gcgtcagccg ggcaggatag
     5281 gtgaagtagg cccacccgcg agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg
     5341 cggtgctcaa cgggaatcct gctctgcgag gctggccgat aagctctgat accgctcgcc
     5401 gcagccgaac gaccgagcgc agcgggtcag tgagcgagga agcggaagag cgcccaatac
     5461 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagcgcccaa acataacagg
     5521 aagaaaaatg ccccgctgtg ggcggacaaa atagttggga actgggaggg gtggaaatgg
     5581 agtttttaag gattatttag ggaagagtga caaaatagat gggaactggg tgtagcgtcg
     5641 taagctaata cgaaaattaa aaatgacaaa atagtttgga actagatttc acttatctgg
     5701 ttcttgaggc ggttaaaaga gccgtactct tctccgatgt cgactaggcc atgatgctca
     5761 ttctgtggga ccaaaacgaa aaaacaccct ttcgggtgtc ttttctggaa tttggtaccg
     5821 aggctgcaag tcccattaag gaggcgcgcc ttattacaga tcttcctcag agatgagctt
     5881 ctgttccaga tcttcttctg aaatcaactt ttgttccaga tcttcttcag agatgagttt
     5941 ctgctccgcg gccgcgtcac ctcctagctg actcaaatca atgcgtgttt cataaagacc
     6001 agtgatggat tgatggataa gagtggcatc taaaacttct tttgtagacg tatatcgttt
     6061 acgatcaatt gttgtatcaa aatatttaaa agcagcggga gctccaagat tcgtcaacgt
     6121 aaataaatga ataatatttt ctgcttgttc acgtattggt ttgtctctat gtttgttata
     6181 tgcactaaga actttatcta aattggcatc tgctaaaata acacgcttag aaaattcact
     6241 gatttgctca ataatctcat ctaaataatg cttatgctgc tccacaaaca attgtttttg
     6301 ttcgttatct tctggactac ccttcaactt ttcataatga ctagctaaat ataaaaaatt
     6361 cacatatttg cttggcagag ccagctcatt tcctttttgt aattctccgg cactagccag
     6421 catccgttta cgaccgtttt ctaactcaaa aagactatat ttaggtagtt taatgattaa
     6481 gtctttttta acttccttat atcctttagc ttctaaaaag tcaatcggat ttttttcaaa
     6541 ggaacttctt tccataattg tgatccctag taactcttta acggatttta acttcttcga
     6601 tttccctttt tccaccttag caaccactag gactgaataa gctaccgttg gactatcaaa
     6661 accaccatat ttttttggat cccagtcttt tttacgagca ataagcttgt ccgaatttct
     6721 ttttggtaaa attgactcct tggagaatcc gcctgtctgt acttctgttt tcttgacaat
     6781 attgacttgg ggcatggaca atactttgcg cactgtggca aaatctcgcc ctttatccca
     6841 gacaatttct ccagtttccc cattagtttc gattagaggg cgtttgcgaa tctctccatt
     6901 tgcaagtgta atttctgttt tgaagaagtt catgatatta gagtaaaaga aatattttgc
     6961 ggttgctttg cctatttctt gctcagactt agcaatcatt ttacgaacat cataaacttt
     7021 ataatcacca tagacaaact ccgattcaag ttttggatat ttcttaatca aagcagttcc
     7081 aacgacggca tttagatacg catcatgggc atgatggtaa ttgttaatct cacgtacttt
     7141 atagaattgg aaatcttttc ggaagtcaga aactaattta gattttaagg taatcacttt
     7201 aacctctcga ataagtttat cattttcatc gtatttagta ttcatgcgac tatccaaaat
     7261 ttgtgccaca tgcttagtga tttggcgagt ttcaaccaat tggcgtttga taaaaccagc
     7321 tttatcaagt tcactcaaac ctccacgttc agctttcgtt aaattatcaa acttacgttg
     7381 agtgattaac ttggcgttta gaagttgtct ccaatagttt ttcatctttt tgactacttc
     7441 ttcacttgga acgttatccg atttaccacg atttttatca gaacgcgtta agaccttatt
     7501 gtctattgaa tcgtctttaa ggaaactttg tggaacaatg gcatcgacat cataatcact
     7561 taaacgatta atatctaatt cttggtccac atacatgtct cttccatttt ggagataata
     7621 gagatagagc ttttcatttt gcaattgagt attttcaaca ggatgctctt taagaatctg
     7681 acttcctaat tctttgatac cttcttcgat tcgtttcata cgctctcgcg aatttttctg
     7741 gcccttttga gttgtctgat tttcacgtgc catttcaata acgatatttt ctggcttatg
     7801 ccgccccatt actttgacca attcatcaac aacttttaca gtctgtaaaa tacctttttt
     7861 aatagcaggg ctaccagcta aatttgcaat atgttcatgt aaactatcgc cttgtccaga
     7921 cacttgtgct ttttgaatgt cttctttaaa tgtcaaacta tcatcatgga tcagctgcat
     7981 aaaattgcga ttggcaaaac catctgattt caaaaaatct aatattgttt tgccagattg
     8041 cttatcccta ataccattaa tcaattttcg agacaaacgt ccccaaccag tataacggcg
     8101 acgtttaagc tgtttcatca ccttatcatc aaagaggtga gcatatgttt taagtctttc
     8161 ctcaatcatc tccctatctt caaataaggt caatgttaaa acaatatcct ctaagatatc
     8221 ttcattttct tcattatcca aaaaatcttt atctttaata atttttagca aatcatggta
     8281 ggtacctaat gaagcattaa atctatcttc aactcctgaa atttcaacac tatcaaaaca
     8341 ttctattttt ttgaaataat cttcttttaa ttgcttaacg gttacttttc gatttgtttt
     8401 gaagagtaaa tcaacaatgg ctttcttctg ttcacctgaa agaaatgctg gttttcgcat
     8461 tccttcagta acatatttga cctttgtcaa ttcgttataa accgtaaaat actcataaag
     8521 caaactatgt tttggtagta ctttttcatt tggaagattt ttatcaaagt ttgtcatgcg
     8581 ttcaataaat gattgagctg aagcaccttt atcgacaact tcttcaaaat tccatggggt
     8641 aattgtttct tcagacttcc gagtcatcca tgcaaaacga ctattgccac gcgccaatgg
     8701 accaacataa taagggattc gaaaagtcaa gattttttca atcttctcac gattgtcttt
     8761 taaaaatgga taaaagtctt cttgtcttct caaaatagca tgcagctcac ccaagtgaat
     8821 ttgatgggga atagagccgt tgtcaaaggt ccgttgcttg cgcagcaaat cttcacgatt
     8881 tagtttcacc aataattcct cagtaccatc cattttttct aaaattggtt tgataaattt
     8941 ataaaattct tcttggctag ctcccccatc aatataacct gcatatccgt tttttgattg
     9001 atcaaaaaag atttctttat acttttctgg aagttgttgt cgaactaaag cttttaaaag
     9061 agtcaagtct tgatgatgtt catcgtagcg tttaatcatt gaagctgata ggggagcctt
     9121 agttatttca gtatttactc ttaggatatc tgaaagtaaa atagcatctg ataaattctt
     9181 agctgccaaa aacaaatcag catattgatc tccaatttgc gccaataaat tatctaaatc
     9241 atcatcgtaa gtatcttttg aaagctgtaa tttagcatct tctgccaaat caaaatttga
     9301 tttaaaatta ggggtcaaac ccaatgacaa agcaatgaga ttcccaaata agccattttt
     9361 cttctcaccg gggagctgag caatgagatt ttctaatcgt cttgatttac tcaatcgtgc
     9421 agaaagaatc gctttagcat ctactccact tgcgttaata gggttttctt caaataattg
     9481 attgtaggtt tgtaccaact ggataaatag tttgtccaca tcactattat caggatttaa
     9541 atctccctca atcaaaaaat gaccacgaaa cttaatcata tgcgctaagg ccaaatagat
     9601 taagcgcaaa tccgctttat cagtagaatc taccaatttt tttcgcagat gatagatagt
     9661 tggatatttc tcatgataag caacttcatc tactatattt ccaaaaatag gatgacgttc
     9721 atgcttcttg tcttcttcca ccaaaaaaga ctcttcaagt cgatgaaaga aactatcatc
     9781 tactttcgcc atctcatttg aaaaaatctc ctgtagataa caaatacgat tcttccgacg
     9841 tgtatacctt ctacgagctg tccgtttgag acgagtcgct tccgctgtct ctccactgtc
     9901 aaataaaaga gcccctataa gatttttttt gatactgtgg cggtctgtat ttcccagaac
     9961 cttgaacttt ttagacggaa ccttatattc atcagtgatc accgcccatc cgacgctatt
    10021 tgtgccgata gctaagccta ttgagtattt cttatccata ctagtcctcc ttctcgctca
    10081 attgttatcc gctcacaatt gaatctaagt atcattgtta tccgctcaca agtcaacact
    10141 ctttttgata aattttacta gtctgcagac gtaaaaaaag cggcgtggtt agccgctttt
    10201 ttaattgccg gagcgcaata aaaaagcccc cggaaggtga tcttccgggg gctttctcat
    10261 gcgttgcttg tattgaaaat gggggccatg cctacctatc tgcctcccgg gaagcggcat
    10321 gcatttacgt tgacaccatc gaatggtgca aaacctttcg cggtatggca tgatagcgcc
    10381 cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag
    10441 agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt
    10501 ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc
    10561 gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc
    10621 tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg
    10681 gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg
    10741 tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc
    10801 aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct
    10861 ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg
    10921 tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt
    10981 ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc
    11041 agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc
    11101 aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc
    11161 tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag
    11221 tgggatacga cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac
    11281 aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc
    11341 aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa accaccctgg
    11401 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
    11461 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt aagttagctc
    11521 actcattagg caccccaccc gggtccttga gccccattat cacctccaaa gccttctcgt
    11581 ctggtcagtt tcacctgttt tacgtaaaaa cccgcttcgg cgggttttta cttttgggga
    11641 aacacagaaa aaagcccgca cctgacagtg cgggcttttt ttttcgacca aaggtgcgac
    11701 tactcttgcc tactacctat cgactgagct gaaagaattc gctcactcat taggcacccc
    11761 aggc
//
