LOCUS       sequence-317527-       11761 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Z. mobilis CRISPRi, promoter D, empty guide, sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11761)
  AUTHORS   Banta AB, Enright AL, Siletti C, Peters JM
  TITLE     A High-Efficacy CRISPR Interference System for Gene Function 
            Discovery in Zymomonas mobilis.
  JOURNAL   Appl Environ Microbiol. 2020 Nov 10;86(23). pii: AEM.01621-20. doi: 
            10.1128/AEM.01621-20. Print 2020 Nov 10.
  PUBMED    32978126
REFERENCE   2  (bases 1 to 11761)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl 
            Environ Microbiol."; date: "2020-11-10"; volume: "86(23). pii: 
            AEM.01621-20. doi"; pages: " 10.1128/AEM.01621-20. Print 2020 Nov 
            10"
COMMENT     Sequence Label: pJMP2369
FEATURES             Location/Qualifiers
     source          1..11761
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  152..317
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     CDS             complement(472..561)
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     CDS             complement(join(571..2154,2155..2157,2158..4644,4645..4647,
                     4648..4674))
                     /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    4697..4713
                     /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    4730..4746
                     /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        4947..5024
                     /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             5025..6107
                     /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(5044..5063)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    6384..6400
                     /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(6776..6778,6779..7492)
                     /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             6776..7489
                     /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(6806..6834)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     7429..7449
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    complement(7634..7681)
                     /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(7758..7763,7764..7780,7781..7786)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     primer_bind     complement(7935..7954)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    complement(8618..8665)
                     /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(8716..8802)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(8905..8999)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(9027..9043)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9028..9044)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(9458..9477)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9577..9599
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9637..9655)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9723..9827
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9828..9896,9897..10688)
                     /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(10046..10065)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10989..11377
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            11646..11754
                     /note="incP origin of transfer"
ORIGIN
        1 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcgg gtcagtgagc gaggaagcgg
       61 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagcg
      121 cccaaacata acaggaagaa aaatgccccg ctgtgggcgg acaaaatagt tgggaactgg
      181 gaggggtgga aatggagttt ttaaggatta tttagggaag agtgacaaaa tagatgggaa
      241 ctgggtgtag cgtcgtaagc taatacgaaa attaaaaatg acaaaatagt ttggaactag
      301 atttcactta tctggttctt gaggcggtta aaagagccgt actcttctcc gatgtcgact
      361 aggccatgat gctcattctg tgggaccaaa acgaaaaaac accctttcgg gtgtcttttc
      421 tggaatttgg taccgaggct gcaagtccca ttaaggaggc gcgccttatt acagatcttc
      481 ctcagagatg agcttctgtt ccagatcttc ttctgaaatc aacttttgtt ccagatcttc
      541 ttcagagatg agtttctgct ccgcggccgc gtcacctcct agctgactca aatcaatgcg
      601 tgtttcataa agaccagtga tggattgatg gataagagtg gcatctaaaa cttcttttgt
      661 agacgtatat cgtttacgat caattgttgt atcaaaatat ttaaaagcag cgggagctcc
      721 aagattcgtc aacgtaaata aatgaataat attttctgct tgttcacgta ttggtttgtc
      781 tctatgtttg ttatatgcac taagaacttt atctaaattg gcatctgcta aaataacacg
      841 cttagaaaat tcactgattt gctcaataat ctcatctaaa taatgcttat gctgctccac
      901 aaacaattgt ttttgttcgt tatcttctgg actacccttc aacttttcat aatgactagc
      961 taaatataaa aaattcacat atttgcttgg cagagccagc tcatttcctt tttgtaattc
     1021 tccggcacta gccagcatcc gtttacgacc gttttctaac tcaaaaagac tatatttagg
     1081 tagtttaatg attaagtctt ttttaacttc cttatatcct ttagcttcta aaaagtcaat
     1141 cggatttttt tcaaaggaac ttctttccat aattgtgatc cctagtaact ctttaacgga
     1201 ttttaacttc ttcgatttcc ctttttccac cttagcaacc actaggactg aataagctac
     1261 cgttggacta tcaaaaccac catatttttt tggatcccag tcttttttac gagcaataag
     1321 cttgtccgaa tttctttttg gtaaaattga ctccttggag aatccgcctg tctgtacttc
     1381 tgttttcttg acaatattga cttggggcat ggacaatact ttgcgcactg tggcaaaatc
     1441 tcgcccttta tcccagacaa tttctccagt ttccccatta gtttcgatta gagggcgttt
     1501 gcgaatctct ccatttgcaa gtgtaatttc tgttttgaag aagttcatga tattagagta
     1561 aaagaaatat tttgcggttg ctttgcctat ttcttgctca gacttagcaa tcattttacg
     1621 aacatcataa actttataat caccatagac aaactccgat tcaagttttg gatatttctt
     1681 aatcaaagca gttccaacga cggcatttag atacgcatca tgggcatgat ggtaattgtt
     1741 aatctcacgt actttataga attggaaatc ttttcggaag tcagaaacta atttagattt
     1801 taaggtaatc actttaacct ctcgaataag tttatcattt tcatcgtatt tagtattcat
     1861 gcgactatcc aaaatttgtg ccacatgctt agtgatttgg cgagtttcaa ccaattggcg
     1921 tttgataaaa ccagctttat caagttcact caaacctcca cgttcagctt tcgttaaatt
     1981 atcaaactta cgttgagtga ttaacttggc gtttagaagt tgtctccaat agtttttcat
     2041 ctttttgact acttcttcac ttggaacgtt atccgattta ccacgatttt tatcagaacg
     2101 cgttaagacc ttattgtcta ttgaatcgtc tttaaggaaa ctttgtggaa caatggcatc
     2161 gacatcataa tcacttaaac gattaatatc taattcttgg tccacataca tgtctcttcc
     2221 attttggaga taatagagat agagcttttc attttgcaat tgagtatttt caacaggatg
     2281 ctctttaaga atctgacttc ctaattcttt gataccttct tcgattcgtt tcatacgctc
     2341 tcgcgaattt ttctggccct tttgagttgt ctgattttca cgtgccattt caataacgat
     2401 attttctggc ttatgccgcc ccattacttt gaccaattca tcaacaactt ttacagtctg
     2461 taaaatacct tttttaatag cagggctacc agctaaattt gcaatatgtt catgtaaact
     2521 atcgccttgt ccagacactt gtgctttttg aatgtcttct ttaaatgtca aactatcatc
     2581 atggatcagc tgcataaaat tgcgattggc aaaaccatct gatttcaaaa aatctaatat
     2641 tgttttgcca gattgcttat ccctaatacc attaatcaat tttcgagaca aacgtcccca
     2701 accagtataa cggcgacgtt taagctgttt catcacctta tcatcaaaga ggtgagcata
     2761 tgttttaagt ctttcctcaa tcatctccct atcttcaaat aaggtcaatg ttaaaacaat
     2821 atcctctaag atatcttcat tttcttcatt atccaaaaaa tctttatctt taataatttt
     2881 tagcaaatca tggtaggtac ctaatgaagc attaaatcta tcttcaactc ctgaaatttc
     2941 aacactatca aaacattcta tttttttgaa ataatcttct tttaattgct taacggttac
     3001 ttttcgattt gttttgaaga gtaaatcaac aatggctttc ttctgttcac ctgaaagaaa
     3061 tgctggtttt cgcattcctt cagtaacata tttgaccttt gtcaattcgt tataaaccgt
     3121 aaaatactca taaagcaaac tatgttttgg tagtactttt tcatttggaa gatttttatc
     3181 aaagtttgtc atgcgttcaa taaatgattg agctgaagca cctttatcga caacttcttc
     3241 aaaattccat ggggtaattg tttcttcaga cttccgagtc atccatgcaa aacgactatt
     3301 gccacgcgcc aatggaccaa cataataagg gattcgaaaa gtcaagattt tttcaatctt
     3361 ctcacgattg tcttttaaaa atggataaaa gtcttcttgt cttctcaaaa tagcatgcag
     3421 ctcacccaag tgaatttgat ggggaataga gccgttgtca aaggtccgtt gcttgcgcag
     3481 caaatcttca cgatttagtt tcaccaataa ttcctcagta ccatccattt tttctaaaat
     3541 tggtttgata aatttataaa attcttcttg gctagctccc ccatcaatat aacctgcata
     3601 tccgtttttt gattgatcaa aaaagatttc tttatacttt tctggaagtt gttgtcgaac
     3661 taaagctttt aaaagagtca agtcttgatg atgttcatcg tagcgtttaa tcattgaagc
     3721 tgatagggga gccttagtta tttcagtatt tactcttagg atatctgaaa gtaaaatagc
     3781 atctgataaa ttcttagctg ccaaaaacaa atcagcatat tgatctccaa tttgcgccaa
     3841 taaattatct aaatcatcat cgtaagtatc ttttgaaagc tgtaatttag catcttctgc
     3901 caaatcaaaa tttgatttaa aattaggggt caaacccaat gacaaagcaa tgagattccc
     3961 aaataagcca tttttcttct caccggggag ctgagcaatg agattttcta atcgtcttga
     4021 tttactcaat cgtgcagaaa gaatcgcttt agcatctact ccacttgcgt taatagggtt
     4081 ttcttcaaat aattgattgt aggtttgtac caactggata aatagtttgt ccacatcact
     4141 attatcagga tttaaatctc cctcaatcaa aaaatgacca cgaaacttaa tcatatgcgc
     4201 taaggccaaa tagattaagc gcaaatccgc tttatcagta gaatctacca atttttttcg
     4261 cagatgatag atagttggat atttctcatg ataagcaact tcatctacta tatttccaaa
     4321 aataggatga cgttcatgct tcttgtcttc ttccaccaaa aaagactctt caagtcgatg
     4381 aaagaaacta tcatctactt tcgccatctc atttgaaaaa atctcctgta gataacaaat
     4441 acgattcttc cgacgtgtat accttctacg agctgtccgt ttgagacgag tcgcttccgc
     4501 tgtctctcca ctgtcaaata aaagagcccc tataagattt tttttgatac tgtggcggtc
     4561 tgtatttccc agaaccttga actttttaga cggaacctta tattcatcag tgatcaccgc
     4621 ccatccgacg ctatttgtgc cgatagctaa gcctattgag tatttcttat ccatactagt
     4681 cctccttctc gctcaattgt tatccgctca caattgaatc taagtatcat tgttatccgc
     4741 tcacaagtca acactctttt tgataaattt tactagtctg cagacgtaaa aaaagcggcg
     4801 tggttagccg cttttttaat tgccggagcg caataaaaaa gcccccggaa ggtgatcttc
     4861 cgggggcttt ctcatgcgtt gcttgtattg aaaatggggg ccatgcctac ctatctgcct
     4921 cccgggaagc ggcatgcatt tacgttgaca ccatcgaatg gtgcaaaacc tttcgcggta
     4981 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     5041 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     5101 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     5161 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     5221 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     5281 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     5341 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     5401 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     5461 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
     5521 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     5581 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     5641 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     5701 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     5761 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     5821 cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
     5881 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     5941 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     6001 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6061 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     6121 aatgtaagtt agctcactca ttaggcaccc cacccgggtc cttgagcccc attatcacct
     6181 ccaaagcctt ctcgtctggt cagtttcacc tgttttacgt aaaaacccgc ttcggcgggt
     6241 ttttactttt ggggaaacac agaaaaaagc ccgcacctga cagtgcgggc tttttttttc
     6301 gaccaaaggt gcgactactc ttgcctacta cctatcgact gagctgaaag aattcggaaa
     6361 attttttttc aaaagtactt gaattgtgag cggataacaa ttataattct agtagagacc
     6421 aactttggtc tccaccatag cggtcggtct ctgtttaaga gctatgctgg aaacagcata
     6481 gcaagtttaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     6541 tttttttgaa ttcatgtggc tgaccgttct gttgtctctc gctcttccga gtagacgaac
     6601 aataaggcct ccctaacggg gggccttttt tattgataac aaaagtcagt gcttccgcta
     6661 tttccaaaat accgggctaa tacggtttaa acgaaaattt atcaaaaaga gtattgactt
     6721 aaagtctaac ctataggata cttacagcca gatctgagcg agaaggaggt aaagtatgag
     6781 caaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt
     6841 taatgggcac aaattttctg tccgtggaga gggtgaaggt gatgctacaa acggaaaact
     6901 cacccttaaa tttatttgca ctactggaaa actacctgtt ccgtggccaa cacttgtcac
     6961 tactctgacc tatggtgttc aatgcttttc ccgttatccg gatcacatga aacggcatga
     7021 ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa cgcactatat ctttcaaaga
     7081 tgacgggacc tacaagacgc gtgctgaagt caagtttgaa ggtgataccc ttgttaatcg
     7141 tatcgagtta aagggtattg attttaaaga agatggaaac attcttggac acaaactgga
     7201 gtacaacttt aactcacaca atgtatacat cacggcagac aaacaaaaga atggaatcaa
     7261 agctaacttc aaaattcgcc acaacgttga agatggttcc gttcaactag cagaccatta
     7321 tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc
     7381 gacacaatct gtcctttcga aagatcccaa cgaaaagcgt gaccacatgg tccttcttga
     7441 gtttgtaact gctgctggga ttacacatgg catggatgag ctctacaaat aataagttta
     7501 aacgacctcc tggatttgct cagacagcct tttcgtcatt cgtttcagcc aaaaaactta
     7561 agaccgccgg tcttgtccac taccttgcag taatgcggtg gacaggatcg gcggttttct
     7621 tttctcttct caagaagttc ctatactttc tagagaatag gaacttcgga ataggaactt
     7681 cctcctgaac ggccataaga acgaaggctg tctgttgaac tctcgagccg tggaaacgga
     7741 tgaaggcacg aacccagttg acataagcct gttcggttcg taaactgtaa tgcaagtagc
     7801 gtatgcgctc atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg
     7861 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
     7921 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
     7981 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
     8041 cctttattca cattcttgcc cgcctgatga atgctcatcc ggagttccgt atggcaatga
     8101 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
     8161 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
     8221 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
     8281 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
     8341 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
     8401 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
     8461 gcttccatgt cggcagaatg cttaatgaat tacaacagtt tttatgcatg cgcccaatac
     8521 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     8581 ccgactggaa ctcgagcatc agcttcaaaa gcgctctgaa gttcctatac tttctagaga
     8641 ataggaactt cggaataggt acttcaagat ccccaattcg agctcatcgt ccgggccgca
     8701 agctcctagc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
     8761 acgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgcctca agctagagag
     8821 tcattacccc aggcgtttaa gggcaccaat aactgcctta aaaaaattac gccccgccct
     8881 gccactcatc gcagtctagc ttggattctc accaataaaa aacgcccggc ggcaaccgag
     8941 cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc
     9001 aagctcagct aattaagcta gcttatcgat accgtcgacc tcgaacccca cgcccctctt
     9061 taatacgacg ggcaatttgc acttcagaaa atgaagagtt tgctttagcc ataacaaaag
     9121 tccagtatgc tttttcacag cataactgga ctgatttcag tttacaacta ttctgtctag
     9181 tttaagactt tattgtcata gtttagatct attttgttca gtttaagact ttattgtccg
     9241 cccacacctg cttacgcagg gcatccattt attactcaac cgtaaccgat tttgccaggt
     9301 tacgcggctg gtcctctagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     9361 aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc
     9421 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     9481 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     9541 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     9601 tttcaccgtc gtcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     9661 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     9721 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     9781 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     9841 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     9901 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     9961 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
    10021 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
    10081 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
    10141 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
    10201 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
    10261 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
    10321 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
    10381 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
    10441 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
    10501 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtcccgc ggtatcattg
    10561 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
    10621 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
    10681 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
    10741 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
    10801 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggcagttatt
    10861 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
    10921 attgatttaa aacttcattt ttaatttttg cggccgcaag atccggccac gatgcgtccg
    10981 gcgtagagga tctgaagatc agcagttcaa cctgttgata gtacgtacta agctctcatg
    11041 tttcacgtac taagctctca tgtttaacgt actaagctct catgtttaac gaactaaacc
    11101 ctcatggcta acgtactaag ctctcatggc taacgtacta agctctcatg tttcacgtac
    11161 taagctctca tgtttgaaca ataaaattaa tataaatcag caacttaaat agcctctaag
    11221 gttttaagtt ttataagaaa aaaaagaata tataaggctt ttaaagcttt taaggtttaa
    11281 cggttgtgga caacaagcca gggatgtaac gcactgagaa gcccttagag cctctcaaag
    11341 caattttgag tgacacagga acacttaacg gctgacatgg gaattagcca tggcatcaca
    11401 gtatcgtgat gacagaggca gggagtgcgg cctttttacg gttcctggcc ttttgctggc
    11461 cttttgctca catgttcttt cctgcgttat caggggattc cttaaggtat actttccgct
    11521 gcataaccct gcttcggggt cattatagcg attttttcgg tatatccatc ctttttcgca
    11581 cgatatacag gattttgcca aagggttcgt gtagactttc cttggtgtat ccaacggcgt
    11641 cagccgggca ggataggtga agtaggccca cccgcgagcg ggtgttcctt cttcactgtc
    11701 ccttattcgc acctggcggt gctcaacggg aatcctgctc tgcgaggctg gccgataagc
    11761 t
//
