LOCUS       sequence-317536-       11858 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 11858)
  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 11858)
  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: pJMP2754
FEATURES             Location/Qualifiers
     source          1..11858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(135..182)
                     /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(299..304,305..321,322..327)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             516..1049
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    complement(1097..1144)
                     /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(1195..1281)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(1384..1478)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1506..1522)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1507..1523)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1937..1956)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2056..2078
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2116..2134)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2202..2306
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2307..2375,2376..3167)
                     /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(2525..2544)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3468..3856
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            4125..4233
                     /note="incP origin of transfer"
     mobile_element  4337..4502
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     CDS             complement(4657..4746)
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             complement(join(4756..6339,6340..6342,6343..8829,8830..8832,
                     8833..8859))
                     /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    8901..8917
                     /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        9299..9376
                     /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             9377..10459
                     /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(9396..9415)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    10744..10760
                     /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(11135..11137,11138..11851)
                     /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             11135..11848
                     /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(11165..11193)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     11788..11808
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 aaacgacctc ctggatttgc tcagacagcc ttttcgtcat tcgtttcagc caaaaaactt
       61 aagaccgccg gtcttgtcca ctaccttgca gtaatgcggt ggacaggatc ggcggttttc
      121 ttttctcttc tcaagaagtt cctatacttt ctagagaata ggaacttcgg aataggaact
      181 tcctcctgaa cggccataag aacgaaggct gtctgttgaa ctctcgagcc ttgtcgcctt
      241 gcgtataata tttgcccatg gacgcacacc gtggaaacgg atgaaggcac gaacccagtt
      301 gacataagcc tgttcggttc gtaaactgta atgcaagtag cgtatgcgct cacgcaactg
      361 gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg
      421 ttatgactgt ttttttgtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc
      481 cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagc aacgatgtta
      541 cgcagcaggg cagtcgccct aaaacaaagt taggtggctc aagtatgggc atcattcgca
      601 catgtaggct cggccctgac caagtcaaat ccatgcgggc tgctcttgat cttttcggtc
      661 gtgagttcgg agacgtagcc acctactccc aacatcagcc ggactccgat tacctcggga
      721 acttgctccg tagtaagaca ttcatcgcgc ttgctgcctt cgaccaagaa gcggttgttg
      781 gcgctctcgc ggcttacgtt ctgcccaagt ttgagcagcc gcgtagtgag atctatatct
      841 atgatctcgc agtctccgga gagcaccgga ggcagggcat tgccaccgcg ctcatcaatc
      901 tcctcaagca tgaggccaac gcgcttggtg cttatgtgat ctacgtgcaa gcagattacg
      961 gtgacgatcc cgcagtggct ctctatacaa agttgggcat acgggaagaa gtgatgcact
     1021 ttgatatcga cccaagtacc gccacctaac aattcgttca agccgagatc tcgagcatca
     1081 gcttcaaaag cgctctgaag ttcctatact ttctagagaa taggaacttc ggaataggta
     1141 cttcaagatc cccaattcga gctcatcgtc cgggccgcaa gctcctagcg gcggatttgt
     1201 cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga
     1261 ctgagccttt cgttttattt gatgcctcaa gctagagagt cattacccca ggcgtttaag
     1321 ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg cagtctagct
     1381 tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
     1441 gagttctgag gtcattactg gatctatcaa caggagtcca agctcagcta attaagctag
     1501 cttatcgata ccgtcgacct cgaaccccac gcccctcttt aatacgacgg gcaatttgca
     1561 cttcagaaaa tgaagagttt gctttagcca taacaaaagt ccagtatgct ttttcacagc
     1621 ataactggac tgatttcagt ttacaactat tctgtctagt ttaagacttt attgtcatag
     1681 tttagatcta ttttgttcag tttaagactt tattgtccgc ccacacctgc ttacgcaggg
     1741 catccattta ttactcaacc gtaaccgatt ttgccaggtt acgcggctgg tcctctagct
     1801 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     1861 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     1921 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     1981 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     2041 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtcg tcaccgaaac
     2101 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     2161 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2221 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2281 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2341 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2401 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2461 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     2521 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     2581 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2641 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2701 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2761 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2821 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     2881 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2941 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3001 aatctggagc cggtgagcgt gggtcccgcg gtatcattgc agcactgggg ccagatggta
     3061 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3121 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3181 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3241 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3301 gagcgtcaga ccccgtagaa aagatcaaag gcagttattg gtgcctcact gattaagcat
     3361 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3421 taatttttgc ggccgcaaga tccggccacg atgcgtccgg cgtagaggat ctgaagatca
     3481 gcagttcaac ctgttgatag tacgtactaa gctctcatgt ttcacgtact aagctctcat
     3541 gtttaacgta ctaagctctc atgtttaacg aactaaaccc tcatggctaa cgtactaagc
     3601 tctcatggct aacgtactaa gctctcatgt ttcacgtact aagctctcat gtttgaacaa
     3661 taaaattaat ataaatcagc aacttaaata gcctctaagg ttttaagttt tataagaaaa
     3721 aaaagaatat ataaggcttt taaagctttt aaggtttaac ggttgtggac aacaagccag
     3781 ggatgtaacg cactgagaag cccttagagc ctctcaaagc aattttgagt gacacaggaa
     3841 cacttaacgg ctgacatggg aattagccat ggcatcacag tatcgtgatg acagaggcag
     3901 ggagtgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     3961 ctgcgttatc aggggattcc ttaaggtata ctttccgctg cataaccctg cttcggggtc
     4021 attatagcga ttttttcggt atatccatcc tttttcgcac gatatacagg attttgccaa
     4081 agggttcgtg tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa
     4141 gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg
     4201 ctcaacggga atcctgctct gcgaggctgg ccgataagct ctgataccgc tcgccgcagc
     4261 cgaacgaccg agcgcagcgg gtcagtgagc gaggaagcgg aagagcccaa acataacagg
     4321 aagaaaaatg ccccgctgtg ggcggacaaa atagttggga actgggaggg gtggaaatgg
     4381 agtttttaag gattatttag ggaagagtga caaaatagat gggaactggg tgtagcgtcg
     4441 taagctaata cgaaaattaa aaatgacaaa atagtttgga actagatttc acttatctgg
     4501 ttcttgaggc ggttaaaaga gccgtactct tctccgatgt cgactaggcc atgatgctca
     4561 ttctgtggga ccaaaacgaa aaaacaccct ttcgggtgtc ttttctggaa tttggtaccg
     4621 aggctgcaag tcccattaag gaggcgcgcc ttattacaga tcttcctcag agatgagctt
     4681 ctgttccaga tcttcttctg aaatcaactt ttgttccaga tcttcttcag agatgagttt
     4741 ctgctccgcg gccgcgtcac ctcctagctg actcaaatca atgcgtgttt cataaagacc
     4801 agtgatggat tgatggataa gagtggcatc taaaacttct tttgtagacg tatatcgttt
     4861 acgatcaatt gttgtatcaa aatatttaaa agcagcggga gctccaagat tcgtcaacgt
     4921 aaataaatga ataatatttt ctgcttgttc acgtattggt ttgtctctat gtttgttata
     4981 tgcactaaga actttatcta aattggcatc tgctaaaata acacgcttag aaaattcact
     5041 gatttgctca ataatctcat ctaaataatg cttatgctgc tccacaaaca attgtttttg
     5101 ttcgttatct tctggactac ccttcaactt ttcataatga ctagctaaat ataaaaaatt
     5161 cacatatttg cttggcagag ccagctcatt tcctttttgt aattctccgg cactagccag
     5221 catccgttta cgaccgtttt ctaactcaaa aagactatat ttaggtagtt taatgattaa
     5281 gtctttttta acttccttat atcctttagc ttctaaaaag tcaatcggat ttttttcaaa
     5341 ggaacttctt tccataattg tgatccctag taactcttta acggatttta acttcttcga
     5401 tttccctttt tccaccttag caaccactag gactgaataa gctaccgttg gactatcaaa
     5461 accaccatat ttttttggat cccagtcttt tttacgagca ataagcttgt ccgaatttct
     5521 ttttggtaaa attgactcct tggagaatcc gcctgtctgt acttctgttt tcttgacaat
     5581 attgacttgg ggcatggaca atactttgcg cactgtggca aaatctcgcc ctttatccca
     5641 gacaatttct ccagtttccc cattagtttc gattagaggg cgtttgcgaa tctctccatt
     5701 tgcaagtgta atttctgttt tgaagaagtt catgatatta gagtaaaaga aatattttgc
     5761 ggttgctttg cctatttctt gctcagactt agcaatcatt ttacgaacat cataaacttt
     5821 ataatcacca tagacaaact ccgattcaag ttttggatat ttcttaatca aagcagttcc
     5881 aacgacggca tttagatacg catcatgggc atgatggtaa ttgttaatct cacgtacttt
     5941 atagaattgg aaatcttttc ggaagtcaga aactaattta gattttaagg taatcacttt
     6001 aacctctcga ataagtttat cattttcatc gtatttagta ttcatgcgac tatccaaaat
     6061 ttgtgccaca tgcttagtga tttggcgagt ttcaaccaat tggcgtttga taaaaccagc
     6121 tttatcaagt tcactcaaac ctccacgttc agctttcgtt aaattatcaa acttacgttg
     6181 agtgattaac ttggcgttta gaagttgtct ccaatagttt ttcatctttt tgactacttc
     6241 ttcacttgga acgttatccg atttaccacg atttttatca gaacgcgtta agaccttatt
     6301 gtctattgaa tcgtctttaa ggaaactttg tggaacaatg gcatcgacat cataatcact
     6361 taaacgatta atatctaatt cttggtccac atacatgtct cttccatttt ggagataata
     6421 gagatagagc ttttcatttt gcaattgagt attttcaaca ggatgctctt taagaatctg
     6481 acttcctaat tctttgatac cttcttcgat tcgtttcata cgctctcgcg aatttttctg
     6541 gcccttttga gttgtctgat tttcacgtgc catttcaata acgatatttt ctggcttatg
     6601 ccgccccatt actttgacca attcatcaac aacttttaca gtctgtaaaa tacctttttt
     6661 aatagcaggg ctaccagcta aatttgcaat atgttcatgt aaactatcgc cttgtccaga
     6721 cacttgtgct ttttgaatat cttctttaaa tgtcaaacta tcatcatgga tcagctgcat
     6781 aaaattgcga ttggcaaaac catctgattt caaaaaatct aatattgttt tgccagattg
     6841 cttatcccta ataccattaa tcaattttcg agacaaacgt ccccaaccag tataacggcg
     6901 acgtttaagc tgtttcatca ccttatcatc aaagaggtga gcatatgttt taagtctttc
     6961 ctcaatcatc tccctatctt caaataaggt caatgttaaa acaatatcct ctaagatatc
     7021 ttcattttct tcattatcca aaaaatcttt atctttaata atttttagca aatcatggta
     7081 ggtacctaat gaagcattaa atctatcttc aactcctgaa atttcaacac tatcaaaaca
     7141 ttctattttt ttgaaataat cttcttttaa ttgcttaacg gttacttttc gatttgtttt
     7201 gaagagtaaa tcaacaatgg ctttcttctg ttcacctgaa agaaatgctg gttttcgcat
     7261 tccttcagta acatatttga cctttgtcaa ttcgttataa accgtaaaat actcataaag
     7321 caaactatgt tttggtagta ctttttcatt tggaagattt ttatcaaagt ttgtcatgcg
     7381 ttcaataaat gattgagctg aagcaccttt atcgacaact tcttcaaaat tccatggggt
     7441 aattgtttct tcagacttcc gagtcatcca tgcaaaacga ctattgccac gcgccaatgg
     7501 accaacataa taagggattc gaaaagtcaa gattttttca atcttctcac gattgtcttt
     7561 taaaaatgga taaaaatctt cttgtcgtct caaaatagca tgcagctcac ccaagtgaat
     7621 ttgatgggga atagagccgt tgtcaaaggt ccgttgcttg cgcagcaaat cttcacgatt
     7681 tagtttcacc aataattcct cagtaccatc cattttttct aaaattggtt tgataaattt
     7741 ataaaattct tcttggctag ctcccccatc aatataacct gcatatccgt tttttgattg
     7801 atcaaaaaag atttctttat acttttctgg aagttgttgt cgaactaaag cttttaaaag
     7861 agtcaagtct tgatgatgtt catcgtagcg tttaatcatt gaagctgata ggggagcctt
     7921 agttatttca gtatttactc ttaggatatc tgaaagtaaa atagcatctg ataaattctt
     7981 agctgccaaa aacaaatcag catattgatc tccaatttgc gccaataaat tatctaaatc
     8041 atcatcgtaa gtatcttttg aaagctgtaa tttagcatct tctgccaaat caaaatttga
     8101 tttaaaatta ggggtcaaac ccaatgacaa agcaatgaga ttcccaaata agccattttt
     8161 cttctcaccg gggagctgag caatgagatt ttctaatcgt cttgatttac tcaatcgtgc
     8221 agaaagaatc gctttagcat ctactccact tgcgttaata gggttttctt caaataattg
     8281 attgtaggtt tgtaccaact ggataaatag tttgtccaca tcactattat caggatttaa
     8341 atctccctca atcaaaaaat gaccacgaaa cttaatcata tgcgctaagg ccaaatagat
     8401 taagcgcaaa tccgctttat cagtagaatc taccaatttt tttcgcagat gatagatagt
     8461 tggatatttc tcatgataag caacttcatc tactatattt ccaaaaatag gatgacgttc
     8521 atgcttctta tcttcttcca ccaaaaaaga ctcttcaagt cgatgaaaga aactatcatc
     8581 tactttcgcc atctcatttg aaaaaatctc ctgtagataa caaatacgat tcttccgacg
     8641 tgtatacctt ctacgagctg tccgtttgag acgagtcgct tccgctgtct ctccactgtc
     8701 aaataaaaga gcccctataa gatttttttt gatactgtgg cggtctgtat ttcccagaac
     8761 cttgaacttt ttagacggaa ccttatattc atcagtgatc accgcccatc cgacgctatt
     8821 tgtgccgata gctaagccta ttgagtattt cttatccata ctagttttcc tgtgtgaaac
     8881 ctgctgatgt gctcagtatc ttgttatccg ctcacaatgt caatgttatc cgctcacatt
     8941 tataatattt tatctgatta ataagatgat cttcttgaga tcgttttggt ctgcgcgtaa
     9001 tctcttgctc tgaaaacgaa aaaaccgcct tgcagggcgg tttttcgaag gttctctgag
     9061 ctaccaactc tttgaaccga ggtaactggc ttggtgataa gctgtcaaac cagatcaatt
     9121 cgcactagtc tgcagacgta aaaaaagcgg cgtggttagc cgctttttta attgccggag
     9181 cgcaataaaa aagcccccgg aaggtgatct tccgggggct ttctcatgcg ttgcttgtat
     9241 tgaaaatggg ggccatgcct acctatctgc ctcccgggaa gcggcatgca tttacgttga
     9301 caccatcgaa tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
     9361 attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt
     9421 ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg
     9481 ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca
     9541 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc
     9601 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt
     9661 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct
     9721 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc
     9781 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc
     9841 catcaacagt attattttct cccatgaaga tggtacgcga ctgggcgtgg agcatctggt
     9901 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg
     9961 tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga
    10021 acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga
    10081 gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg
    10141 cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga
    10201 taccgaagat agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct
    10261 gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg
    10321 caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca
    10381 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
    10441 actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac
    10501 cccacccggg tccttgagcc ccattatcac ctccaaagcc ttctcgtctg gtcagtttca
    10561 cctgttttac gtaaaaaccc gcttcggcgg gtttttactt ttggggaaac acagaaaaaa
    10621 gcccgcacct gacagtgcgg gctttttttt tcgaccaaag gtgcgactac tcttgcctac
    10681 tacctatcga ctgagctgaa agaattccta aagatctata aatgtgagcg gataacattg
    10741 acattgtgag cggataacaa gatacttcta gttgagacca actttggtct ccaccatagc
    10801 ggtcggtctc tgtttaagag ctatgctgga aacagcatag caagtttaaa taaggctagt
    10861 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttttgaat tcatgtggct
    10921 gaccgttctg ttgtctctcg ctcttccgag tagacgaaca ataaggcctc cctaacgggg
    10981 ggcctttttt attgataaca aaagtcagtg cttccgctat ttccaaaata ccgggctaat
    11041 acggtttaaa cgaaaattta tcaaaaagag tattgactta aagtctaacc tataggatac
    11101 ttacagccag atctgagcga gaaggaggta aagtatgagc aaaggagaag aacttttcac
    11161 tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt
    11221 ccgtggagag ggtgaaggtg atgctacaaa cggaaaactc acccttaaat ttatttgcac
    11281 tactggaaaa ctacctgttc cgtggccaac acttgtcact actctgacct atggtgttca
    11341 atgcttttcc cgttatccgg atcacatgaa acggcatgac tttttcaaga gtgccatgcc
    11401 cgaaggttat gtacaggaac gcactatatc tttcaaagat gacgggacct acaagacgcg
    11461 tgctgaagtc aagtttgaag gtgataccct tgttaatcgt atcgagttaa agggtattga
    11521 ttttaaagaa gatggaaaca ttcttggaca caaactggag tacaacttta actcacacaa
    11581 tgtatacatc acggcagaca aacaaaagaa tggaatcaaa gctaacttca aaattcgcca
    11641 caacgttgaa gatggttccg ttcaactagc agaccattat caacaaaata ctccaattgg
    11701 cgatggccct gtccttttac cagacaacca ttacctgtcg acacaatctg tcctttcgaa
    11761 agatcccaac gaaaagcgtg accacatggt ccttcttgag tttgtaactg ctgctgggat
    11821 tacacatggc atggatgagc tctacaaata ataagttt
//
