LOCUS       sequence-317532-       11762 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Z. mobilis CRISPRi, promoter E, empty guide, sfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11762)
  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 11762)
  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: pJMP2371
FEATURES             Location/Qualifiers
     source          1..11762
                     /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..6403
                     /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."
     CDS             join(6777..6779,6780..7493)
                     /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             6777..7490
                     /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(6807..6835)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     7430..7450
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    complement(7635..7682)
                     /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(7759..7764,7765..7781,7782..7787)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     primer_bind     complement(7936..7955)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    complement(8619..8666)
                     /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(8717..8803)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(8906..9000)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(9028..9044)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9029..9045)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(9459..9478)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9578..9600
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9638..9656)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9724..9828
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9829..9897,9898..10689)
                     /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(10047..10066)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10990..11378
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            11647..11755
                     /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 tacaattgtg agcgctcaca attataattc tagtagagac
     6421 caactttggt ctccaccata gcggtcggtc tctgtttaag agctatgctg gaaacagcat
     6481 agcaagttta aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
     6541 ttttttttga attcatgtgg ctgaccgttc tgttgtctct cgctcttccg agtagacgaa
     6601 caataaggcc tccctaacgg ggggcctttt ttattgataa caaaagtcag tgcttccgct
     6661 atttccaaaa taccgggcta atacggttta aacgaaaatt tatcaaaaag agtattgact
     6721 taaagtctaa cctataggat acttacagcc agatctgagc gagaaggagg taaagtatga
     6781 gcaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg
     6841 ttaatgggca caaattttct gtccgtggag agggtgaagg tgatgctaca aacggaaaac
     6901 tcacccttaa atttatttgc actactggaa aactacctgt tccgtggcca acacttgtca
     6961 ctactctgac ctatggtgtt caatgctttt cccgttatcc ggatcacatg aaacggcatg
     7021 actttttcaa gagtgccatg cccgaaggtt atgtacagga acgcactata tctttcaaag
     7081 atgacgggac ctacaagacg cgtgctgaag tcaagtttga aggtgatacc cttgttaatc
     7141 gtatcgagtt aaagggtatt gattttaaag aagatggaaa cattcttgga cacaaactgg
     7201 agtacaactt taactcacac aatgtataca tcacggcaga caaacaaaag aatggaatca
     7261 aagctaactt caaaattcgc cacaacgttg aagatggttc cgttcaacta gcagaccatt
     7321 atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac cattacctgt
     7381 cgacacaatc tgtcctttcg aaagatccca acgaaaagcg tgaccacatg gtccttcttg
     7441 agtttgtaac tgctgctggg attacacatg gcatggatga gctctacaaa taataagttt
     7501 aaacgacctc ctggatttgc tcagacagcc ttttcgtcat tcgtttcagc caaaaaactt
     7561 aagaccgccg gtcttgtcca ctaccttgca gtaatgcggt ggacaggatc ggcggttttc
     7621 ttttctcttc tcaagaagtt cctatacttt ctagagaata ggaacttcgg aataggaact
     7681 tcctcctgaa cggccataag aacgaaggct gtctgttgaa ctctcgagcc gtggaaacgg
     7741 atgaaggcac gaacccagtt gacataagcc tgttcggttc gtaaactgta atgcaagtag
     7801 cgtatgcgct catcactacc gggcgtattt tttgagttat cgagattttc aggagctaag
     7861 gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat
     7921 cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt
     7981 cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg
     8041 gcctttattc acattcttgc ccgcctgatg aatgctcatc cggagttccg tatggcaatg
     8101 aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag
     8161 caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta
     8221 cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg
     8281 tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat
     8341 ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat
     8401 acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat
     8461 ggcttccatg tcggcagaat gcttaatgaa ttacaacagt ttttatgcat gcgcccaata
     8521 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     8581 cccgactgga actcgagcat cagcttcaaa agcgctctga agttcctata ctttctagag
     8641 aataggaact tcggaatagg tacttcaaga tccccaattc gagctcatcg tccgggccgc
     8701 aagctcctag cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     8761 aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctc aagctagaga
     8821 gtcattaccc caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc
     8881 tgccactcat cgcagtctag cttggattct caccaataaa aaacgcccgg cggcaaccga
     8941 gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc
     9001 caagctcagc taattaagct agcttatcga taccgtcgac ctcgaacccc acgcccctct
     9061 ttaatacgac gggcaatttg cacttcagaa aatgaagagt ttgctttagc cataacaaaa
     9121 gtccagtatg ctttttcaca gcataactgg actgatttca gtttacaact attctgtcta
     9181 gtttaagact ttattgtcat agtttagatc tattttgttc agtttaagac tttattgtcc
     9241 gcccacacct gcttacgcag ggcatccatt tattactcaa ccgtaaccga ttttgccagg
     9301 ttacgcggct ggtcctctag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     9361 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     9421 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     9481 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     9541 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     9601 ttttcaccgt cgtcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     9661 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     9721 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     9781 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     9841 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     9901 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     9961 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
    10021 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
    10081 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
    10141 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
    10201 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
    10261 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
    10321 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
    10381 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
    10441 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
    10501 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtcccg cggtatcatt
    10561 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
    10621 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
    10681 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
    10741 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
    10801 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggcagttat
    10861 tggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
    10921 gattgattta aaacttcatt tttaattttt gcggccgcaa gatccggcca cgatgcgtcc
    10981 ggcgtagagg atctgaagat cagcagttca acctgttgat agtacgtact aagctctcat
    11041 gtttcacgta ctaagctctc atgtttaacg tactaagctc tcatgtttaa cgaactaaac
    11101 cctcatggct aacgtactaa gctctcatgg ctaacgtact aagctctcat gtttcacgta
    11161 ctaagctctc atgtttgaac aataaaatta atataaatca gcaacttaaa tagcctctaa
    11221 ggttttaagt tttataagaa aaaaaagaat atataaggct tttaaagctt ttaaggttta
    11281 acggttgtgg acaacaagcc agggatgtaa cgcactgaga agcccttaga gcctctcaaa
    11341 gcaattttga gtgacacagg aacacttaac ggctgacatg ggaattagcc atggcatcac
    11401 agtatcgtga tgacagaggc agggagtgcg gcctttttac ggttcctggc cttttgctgg
    11461 ccttttgctc acatgttctt tcctgcgtta tcaggggatt ccttaaggta tactttccgc
    11521 tgcataaccc tgcttcgggg tcattatagc gattttttcg gtatatccat cctttttcgc
    11581 acgatataca ggattttgcc aaagggttcg tgtagacttt ccttggtgta tccaacggcg
    11641 tcagccgggc aggataggtg aagtaggccc acccgcgagc gggtgttcct tcttcactgt
    11701 cccttattcg cacctggcgg tgctcaacgg gaatcctgct ctgcgaggct ggccgataag
    11761 ct
//
