LOCUS       sequence-317521-       10953 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Z. mobilis CRISPRi, promoter B, empty guide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10953)
  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 10953)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pJMP2132
FEATURES             Location/Qualifiers
     source          1..10953
                     /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    6385..6404
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    complement(6826..6873)
                     /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(6950..6955,6956..6972,6973..6978)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     primer_bind     complement(7127..7146)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    complement(7810..7857)
                     /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(7908..7994)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(8097..8191)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(8219..8235)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8220..8236)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(8650..8669)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8769..8791
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8829..8847)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8915..9019
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9020..9088,9089..9880)
                     /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(9238..9257)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10181..10569
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            10838..10946
                     /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 aattcgctca
     6361 ctcattaggc accccaggct ttacaattgt gagcgctcac aattataatg tctagttgag
     6421 accaactttg gtctccacca tagcggtcgg tctctgttta agagctatgc tggaaacagc
     6481 atagcaagtt taaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
     6541 cttttttttt gaattcatgt ggctgaccgt tctgttgtct ctcgctcttc cgagtagacg
     6601 aacaataagg cctccctaac ggggggcctt ttttattgat aacaaaagtc agtgcttccg
     6661 ctatttccaa aataccgggc taatacggtt taaacgacct cctggatttg ctcagacagc
     6721 cttttcgtca ttcgtttcag ccaaaaaact taagaccgcc ggtcttgtcc actaccttgc
     6781 agtaatgcgg tggacaggat cggcggtttt cttttctctt ctcaagaagt tcctatactt
     6841 tctagagaat aggaacttcg gaataggaac ttcctcctga acggccataa gaacgaaggc
     6901 tgtctgttga actctcgagc cgtggaaacg gatgaaggca cgaacccagt tgacataagc
     6961 ctgttcggtt cgtaaactgt aatgcaagta gcgtatgcgc tcatcactac cgggcgtatt
     7021 ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg
     7081 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
     7141 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
     7201 cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
     7261 gaatgctcat ccggagttcc gtatggcaat gaaagacggt gagctggtga tatgggatag
     7321 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag
     7381 tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta
     7441 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc
     7501 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
     7561 cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
     7621 ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga
     7681 attacaacag tttttatgca tgcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     7741 gattcattaa tgcagctggc acgacaggtt tcccgactgg aactcgagca tcagcttcaa
     7801 aagcgctctg aagttcctat actttctaga gaataggaac ttcggaatag gtacttcaag
     7861 atccccaatt cgagctcatc gtccgggccg caagctccta gcggcggatt tgtcctactc
     7921 aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttt cgactgagcc
     7981 tttcgtttta tttgatgcct caagctagag agtcattacc ccaggcgttt aagggcacca
     8041 ataactgcct taaaaaaatt acgccccgcc ctgccactca tcgcagtcta gcttggattc
     8101 tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct
     8161 gaggtcatta ctggatctat caacaggagt ccaagctcag ctaattaagc tagcttatcg
     8221 ataccgtcga cctcgaaccc cacgcccctc tttaatacga cgggcaattt gcacttcaga
     8281 aaatgaagag tttgctttag ccataacaaa agtccagtat gctttttcac agcataactg
     8341 gactgatttc agtttacaac tattctgtct agtttaagac tttattgtca tagtttagat
     8401 ctattttgtt cagtttaaga ctttattgtc cgcccacacc tgcttacgca gggcatccat
     8461 ttattactca accgtaaccg attttgccag gttacgcggc tggtcctcta gctggcgtaa
     8521 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     8581 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     8641 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     8701 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     8761 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcgtcaccga aacgcgcgag
     8821 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     8881 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     8941 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     9001 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     9061 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     9121 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     9181 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     9241 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     9301 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     9361 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     9421 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     9481 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     9541 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     9601 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     9661 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     9721 agccggtgag cgtgggtccc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     9781 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     9841 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     9901 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     9961 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
    10021 agaccccgta gaaaagatca aaggcagtta ttggtgcctc actgattaag cattggtaac
    10081 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaatttt
    10141 tgcggccgca agatccggcc acgatgcgtc cggcgtagag gatctgaaga tcagcagttc
    10201 aacctgttga tagtacgtac taagctctca tgtttcacgt actaagctct catgtttaac
    10261 gtactaagct ctcatgttta acgaactaaa ccctcatggc taacgtacta agctctcatg
    10321 gctaacgtac taagctctca tgtttcacgt actaagctct catgtttgaa caataaaatt
    10381 aatataaatc agcaacttaa atagcctcta aggttttaag ttttataaga aaaaaaagaa
    10441 tatataaggc ttttaaagct tttaaggttt aacggttgtg gacaacaagc cagggatgta
    10501 acgcactgag aagcccttag agcctctcaa agcaattttg agtgacacag gaacacttaa
    10561 cggctgacat gggaattagc catggcatca cagtatcgtg atgacagagg cagggagtgc
    10621 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
    10681 atcaggggat tccttaaggt atactttccg ctgcataacc ctgcttcggg gtcattatag
    10741 cgattttttc ggtatatcca tcctttttcg cacgatatac aggattttgc caaagggttc
    10801 gtgtagactt tccttggtgt atccaacggc gtcagccggg caggataggt gaagtaggcc
    10861 cacccgcgag cgggtgttcc ttcttcactg tcccttattc gcacctggcg gtgctcaacg
    10921 ggaatcctgc tctgcgaggc tggccgataa gct
//
