LOCUS       sequence-445118-       14716 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Coxiella burnetii CRISPR-Cas9 cytosine base editing plasmid version 
            1 without sgRNA construct. Expresses 3xF-HF-BE3..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14716)
  AUTHORS   Steiner S, Roy CR
  TITLE     CRISPR-Cas9-based approaches for genetic analysis and epistatic 
            interaction studies in Coxiella burnetii.
  JOURNAL   mSphere. 2024 Nov 19:e0052324. doi: 10.1128/msphere.00523-24.
  PUBMED    39560384
REFERENCE   2  (bases 1 to 14716)
  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: "mSphere. 
            2024 Nov 19:e0052324. doi: 10.1128/msphere.00523-24."
COMMENT     Sequence Label: pCB-CBEv1
FEATURES             Location/Qualifiers
     source          1..14716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      6..33
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        52..143
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(144..212,213..1004)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(362..381)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             1843..2814
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     CDS             3325..4164
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             4151..5002
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(5285..5533)
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             complement(join(5546..9619,9620..9622,9623..9646))
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTAFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFMA
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(9695..10378)
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG
                     RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS
                     RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE
                     AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL
                     WATGLK"
     CDS             complement(10379..10444)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(10465..10487)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10476..10492
                     /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        complement(join(10500..10506,10507..10522,10523..10528))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     promoter        10758..10835
                     /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             10836..11918
                     /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(10855..10874)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     12192..12209
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             12449..13108
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(12515..12534)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      13495..13889
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     oriT            14231..14318
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
ORIGIN
        1 tattgagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat
       61 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
      121 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
      181 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
      241 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
      301 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
      361 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
      421 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
      481 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
      541 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
      601 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
      661 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
      721 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
      781 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
      841 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
      901 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
      961 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaatacgct ggccatccat
     1021 gccggtggcg gcgagaaccc gcactgccac ctgatgatct ccgagcggat caatgacggc
     1081 atcgagcggc ccgccgctca gtggttcaag cggtacaacg gcaagacccc ggagaagggc
     1141 ggggcacaga agaccgaagc gctcaagccc aaggcatggc ttgagcagac ccgcgaggca
     1201 tgggccgacc atgccaaccg ggcattagag cgggctggcc acgacgcccg cattgaccac
     1261 agaacacttg aggcgcaggg catcgagcgc ctgcccggtg ttcacctggg gccgaacgtg
     1321 gtggagatgg aaggccgggg catccgcacc gaccgggcag acgtggccct gaacatcgac
     1381 accgccaacg cccagatcat cgacttacag gaataccggg aggcaataga ccatgaacgc
     1441 aatcgacaga gtgaagaaat ccagaggcat caacgagtta gcggagcaga tcgaaccgct
     1501 ggcccagagc atggcgacac tggccgacga agcccggcag gtcatgagcc agacccagca
     1561 ggccagcgag gcgcaggcgg cggagtggct gaaagcccag cgccagacag gggcggcatg
     1621 ggtggagctg gccaaagagt tgcgggaggt agccgccgag gtgagcagcg ccgcgcagag
     1681 cgcccggagc gcgtcgcggg ggtggcactg gaagctatgg ctaaccgtga tgctggcttc
     1741 catgatgcct acggtggtgc tgctgatcgc atcgttgctc ttgctcgacc tgacgccact
     1801 gacaaccgag gacggctcga tctggctgcg cttggtggcc cgatgaagaa cgacaggact
     1861 ttgcaggcca taggccgaca gctcaaggcc atgggctgtg agcgcttcga tatcggcgtc
     1921 agggacgcca ccaccggcca gatgatgaac cgggaatggt cagccgccga agtgctccag
     1981 aacacgccat ggctcaagcg gatgaatgcc cagggcaatg acgtgtatat caggcccgcc
     2041 gagcaggagc ggcatggtct ggtgctggtg gacgacctca gcgagtttga cctggatgac
     2101 atgaaagccg agggccggga gcctgccctg gtagtggaaa ccagcccgaa gaactatcag
     2161 gcatgggtca aggtggccga cgccgcaggc ggtgaacttc gggggcagat tgcccggacg
     2221 ctggccagcg agtacgacgc cgacccggcc agcgccgaca gccgccacta tggccgcttg
     2281 gcgggcttca ccaaccgcaa ggacaagcac accacccgcg ccggttatca gccgtgggtg
     2341 ctgctgcgtg aatccaaggg caagaccgcc accgctggcc cggcgctggt gcagcaggct
     2401 ggccagcaga tcgagcaggc ccagcggcag caggagaagg cccgcaggct ggccagcctc
     2461 gaactgcccg agcggcagct tagccgccac cggcgcacgg cgctggacga gtaccgcagc
     2521 gagatggccg ggctggtcaa gcgcttcggt gatgacctca gcaagtgcga ctttatcgcc
     2581 gcgcagaagc tggccagccg gggccgcagt gccgaggaaa tcggcaaggc catggccgag
     2641 gccagcccag cgctggcaga gcgcaagccc ggccacgaag cggattacat cgagcgcacc
     2701 gtcagcaagg tcatgggtct gcccagcgtc cagcttgcgc gggccgagct ggcacgggca
     2761 ccggcacccc gccagcgagg catggacagg ggcgggccag atttcagcat gtagtgcttg
     2821 cgttggtact cacgcctgtt atactatgag tactcacgca cagaaggggg ttttatggaa
     2881 tacgaaaaaa gcgcttcagg gtcggtctac ctgatcaaaa gtgacaaggg ctattggttg
     2941 cccggtggct ttggttatac gtcaaacaag gccgaggctg gccgcttttc agtcgctgat
     3001 atggccagcc ttaaccttga cggctgcacc ttgtccttgt tccgcgaaga caagcctttc
     3061 ggccccggca agtttctcgg tgactgatat gaaagaccaa aaggacaagc agaccggcga
     3121 cctgctggcc agccctgacg ctgtacgcca agcgcgatat gccgagcgca tgaaggccaa
     3181 agggatgcgt cagcgcaagt tctggctgac cgacgacgaa tacgaggcgc tgcgcgagtg
     3241 cctggaagaa ctcagagcgg cgcagggcgg gggtagtgac cccgccagcg cctaaccacc
     3301 aactgcctgc aaaggaggca atcaatggct acccataagc ctatcaatat tctggaggcg
     3361 ttcgcagcag cgccgccacc gctggactac gttttgccca acatggtggc cggtacggtc
     3421 ggggcgctgg tgtcgcccgg tggtgccggt aaatccatgc tggccctgca actggccgca
     3481 cagattgcag gcgggccgga tctgctggag gtgggcgaac tgcccaccgg cccggtgatc
     3541 tacctgcccg ccgaagaccc gcccaccgcc attcatcacc gcctgcacgc ccttggggcg
     3601 cacctcagcg ccgaggaacg gcaagccgtg gctgacggcc tgctgatcca gccgctgatc
     3661 ggcagcctgc ccaacatcat ggccccggag tggttcgacg gcctcaagcg cgccgccgag
     3721 ggccgccgcc tgatggtgct ggacacgctg cgccggttcc acatcgagga agaaaacgcc
     3781 agcggcccca tggcccaggt catcggtcgc atggaggcca tcgccgccga taccgggtgc
     3841 tctatcgtgt tcctgcacca tgccagcaag ggcgcggcca tgatgggcgc aggcgaccag
     3901 cagcaggcca gccggggcag ctcggtactg gtcgataaca tccgctggca gtcctacctg
     3961 tcgagcatga ccagcgccga ggccgaggaa tggggtgtgg acgacgacca gcgccggttc
     4021 ttcgtccgct tcggtgtgag caaggccaac tatggcgcac cgttcgctga tcggtggttc
     4081 aggcggcatg acggcggggt gctcaagccc gccgtgctgg agaggcagcg caagagcaag
     4141 ggggtgcccc gtggtgaagc ctaagaacaa gcacagcctc agccacgtcc ggcacgaccc
     4201 ggcgcactgt ctggcccccg gcctgttccg tgccctcaag cggggcgagc gcaagcgcag
     4261 caagctggac gtgacgtatg actacggcga cggcaagcgg atcgagttca gcggcccgga
     4321 gccgctgggc gctgatgatc tgcgcatcct gcaagggctg gtggccatgg ctgggcctaa
     4381 tggcctagtg cttggcccgg aacccaagac cgaaggcgga cggcagctcc ggctgttcct
     4441 ggaacccaag tgggaggccg tcaccgctga tgccatggtg gtcaaaggta gctatcgggc
     4501 gctggcaaag gaaatcgggg cagaggtcga tagtggtggg gcgctcaagc acatacagga
     4561 ctgcatcgag cgcctttgga aggtatccat catcgcccag aatggccgca agcggcaggg
     4621 gtttcggctg ctgtcggagt acgccagcga cgaggcggac gggcgcctgt acgtggccct
     4681 gaaccccttg atcgcgcagg ccgtcatggg tggcggccag catgtgcgca tcagcatgga
     4741 cgaggtgcgg gcgctggaca gcgaaaccgc ccgcctgctg caccagcggc tgtgtggctg
     4801 gatcgacccc ggcaaaaccg gcaaggcttc catagatacc ttgtgcggct atgtctggcc
     4861 gtcagaggcc agtggttcga ccatgcgcaa gcgccgccag cgggtgcgcg aggcgttgcc
     4921 ggagctggtc gcgctgggct ggacggtaac cgagttcgcg gcgggcaagt acgacatcac
     4981 ccggcccaag gcggcaggct gacccccccc actctattgt aaacaagaca tttttatctt
     5041 ttatattcaa tggcttattt tcctgctaat tggtaatacc atgaaaaata ccatgctcag
     5101 aaaaggctta acaatatttt gaaaaattgc ctactgagcg ctgccgcaca gctccatagg
     5161 ccgctttcct ggctttgctt ccagatgtat gctcttctgc tcggtctaga aagtatagga
     5221 acttcagagc gcttttgaag ctaattcgat catgcatgag ctcactagtt taagaaccac
     5281 cagagagcat cttaatcttg ttctcaccgt tgctatcctg tatgaccaga gcccaaggct
     5341 tgtattcagg ggcgtcgcta gtcagaagca tgacattctc gtcggtgctc tcgtcgtagg
     5401 cggtgtgcac gagtatatcg ctttccggct tgttcccaat gacttcttcc acctcctctg
     5461 ggagcatgag gatggattcc tggataacca gttgcttacc ggtctccttt tcaataatat
     5521 ctgacagatt agtagaacca ccagagtcac ctcctagctg actcaaatca atgcgtgttt
     5581 cataaagacc agtgatggat tgatggataa gagtggcatc taaaacttct tttgtagacg
     5641 tatatcgttt acgatcaatt gttgtatcaa aatatttaaa agcagcggga gctccaagat
     5701 tcgtcaacgt aaataaatga ataatatttt ctgcttgttc acgtattggt ttgtctctat
     5761 gtttgttata tgcactaaga actttatcta aattggcatc tgctaaaata acacgcttag
     5821 aaaattcact gatttgctca ataatctcat ctaaataatg cttatgctgc tccacaaaca
     5881 attgtttttg ttcgttatct tctggactac ccttcaactt ttcataatga ctagctaaat
     5941 ataaaaaatt cacatatttg cttggcagag ccagctcatt tcctttttgt aattctccgg
     6001 cactagccag catccgttta cgaccgtttt ctaactcaaa aagactatat ttaggtagtt
     6061 taatgattaa gtctttttta acttccttat atcctttagc ttctaaaaag tcaatcggat
     6121 ttttttcaaa ggaacttctt tccataattg tgatccctag taactcttta acggatttta
     6181 acttcttcga tttccctttt tccaccttag caacaactag gactgaataa gctaccgttg
     6241 gactatcaaa accaccatat ttttttggat cccagtcttt tttacgagca ataagcttgt
     6301 ccgaatttct ttttggtaaa attgactcct tggagaatcc gcctgtctgt acttctgttt
     6361 tcttgacaat attgacttgg ggcatggaca atactttgcg cactgtggca aaatctcgcc
     6421 ctttatccca gacaatttct ccagtttccc cattagtttc gattagaggg cgtttgcgaa
     6481 tctctccatt tgcaagtgta atttctgttt tgaagaagtt catgatatta gagtaaaaga
     6541 aatattttgc ggttgctttg cctatttctt gctcagactt agcaatcatt ttacgaacat
     6601 cataaacttt ataatcacca tagacaaact ccgattcaag ttttggatat ttcttaatca
     6661 aagcagttcc aacgacggca tttagatacg catcatgggc atgatggtaa ttgttaatct
     6721 cacgtacttt atagaattgg aaatcttttc ggaagtcaga aactaattta gattttaagg
     6781 taatcacttt aacctctcga ataagtttat cattttcatc gtatttagta ttcatgcgac
     6841 tatccaaaat ttgtgccaca tgcttagtga ttgcgcgagt ttcaaccaat tggcgtttga
     6901 taaaaccagc tttatcaagt tcactcaaac ctccacgttc agctttcgtt aaattatcaa
     6961 acttacgttg agtgattaac ttggcgttta gaagttgtct ccaatagttt ttcatctttt
     7021 tgactacttc ttcacttgga acgttatccg atttaccacg atttttatca gaacgcgtta
     7081 agaccttatt gtctattgaa tcgtctttaa ggaaactttg tggaacaatg tgatcgacat
     7141 cataatcact taaacgatta atatctaatt cttggtccac atacatgtct cttccatttt
     7201 ggagataata gagatagagc ttttcatttt gcaattgagt attttcaaca ggatgctctt
     7261 taagaatctg acttcctaat tctttgatac cttcttcgat tcgtttcata cgctctcgcg
     7321 aatttttctg gcccttttga gttgtctgat tttcacgtgc catttcaata acgatatttt
     7381 ctggcttatg ccgccccatt actttgacca attcatcaac aacttttaca gtctgtaaaa
     7441 tacctttttt aatagcaggg ctaccagcta aatttgcaat atgttcatgt aaactatcgc
     7501 cttgtccaga cacttgtgct ttttgaatgt cttctttaaa tgtcaaagag tcgtcgtgta
     7561 ttaacgccat aaaattgcga ttggcaaaac catctgattt caaaaaatct aatattgttt
     7621 tgccagattg cttatcccta ataccattaa tcaattttcg agacaaagct ccccaaccag
     7681 tataacggcg acgtttaagc tgtttcatca ccttatcatc aaagaggtga gcatatgttt
     7741 taagtctttc ctcaatcatc tccctatctt caaataaggt caatgttaaa acaatatcct
     7801 ctaagatatc ttcattttct tcattatcca aaaaatcttt atctttaata atttttagca
     7861 aatcatggta ggtacctaat gaagcattaa atctatcttc aactcctgaa atttcaacac
     7921 tatcaaaaca ttctattttt ttgaaataat cttcttttaa ttgcttaacg gttacttttc
     7981 gatttgtttt gaagagtaaa tcaacaatgg ctttcttctg ttcacctgaa agaaatgctg
     8041 gttttcgcat tccttcagta acatatttga cctttgtcaa ttcgttataa accgtaaaat
     8101 actcataaag caaactatgt tttggtagta ctttttcatt tggaagattt ttatcaaagg
     8161 ctgtcatgcg ttcaataaat gattgagctg aagcaccttt atcgacaact tcttcaaaat
     8221 tccatggggt aattgtttct tcagacttcc gagtcatcca tgcaaaacga ctattgccac
     8281 gcgccaatgg accaacataa taaggaattc gaaaagtcaa gattttttca atcttctcac
     8341 gattgtcttt taaaaatgga taaaagtctt cttgtcttct caaaatagca tgcagctcac
     8401 ccaagtgaat ttgatgggga atagagccgt tgtcaaaggt ccgttgcttg cgcagcaaat
     8461 cttcacgatt tagtttcacc aataattcct cagtaccatc cattttttct aaaattggtt
     8521 tgataaattt ataaaattct tcttggctag ctcccccatc aatataacct gcatatccgt
     8581 tttttgattg atcaaaaaag atttctttat acttttctgg aagttgttgt cgaactaaag
     8641 cttttaaaag agtcaagtct tgatgatgtt catcgtagcg tttaatcatt gaagctgata
     8701 ggggagcctt agttatttca gtatttactc ttaggatatc tgaaagtaaa atagcatctg
     8761 ataaattctt agctgccaaa aacaaatcag catattgatc tccaatttgc gccaataaat
     8821 tatctaaatc atcatcgtaa gtatcttttg aaagctgtaa tttagcatct tctgccaaat
     8881 caaaatttga tttaaaatta ggggtcaaac ccaatgacaa agcaatgaga ttcccaaata
     8941 agccattttt cttctcaccg gggagctgag caatgagatt ttctaatcgt cttgatttac
     9001 tcaatcgtgc agaaagaatc gctttagcat ctactccact tgcgttaata gggttttctt
     9061 caaataattg attgtaggtt tgtaccaact ggataaatag tttgtccaca tcactattat
     9121 caggatttaa atctccctca atcaaaaaat gaccacgaaa cttaatcata tgcgctaagg
     9181 ccaaatagat taagcgcaaa tccgctttat cagtagaatc taccaatttt tttcgcagat
     9241 gatagatagt tggatatttc tcatgataag caacttcatc tactatattt ccaaaaatag
     9301 gatgacgttc atgcttcttg tcttcttcca ccaaaaaaga ctcttcaagt cgatgaaaga
     9361 aactatcatc tactttcgcc atctcatttg aaaaaatctc ctgtagataa caaatacgat
     9421 tcttccgacg tgtatacctt ctacgagctg tccgtttgag acgagtcgct tccgctgtct
     9481 ctccactgtc aaataaaaga gcccctataa gatttttttt gatactgtgg cggtctgtat
     9541 ttcccagaac cttgaacttt ttagacggaa ccttatattc atcagtgatc accgcccatc
     9601 cgacgctatt tgtgccgata gctaagccta ttgagtattt cttatcagac tccggggtgg
     9661 ccgattctga agtccccggt gtttcgctac cggatttcag accggtcgcc cacaggatgt
     9721 gcggcggcag acgctggtag tggcaagact gcagcgcgat ggtgaagaag gtcagctgcg
     9781 gctgtttacg acgcaggatg ttcaggcacg gcggcagacc caggatgatg cagtacagtt
     9841 ccagaacgta cagacgaacc cacaggtgcg ggtaacgcgg ccagtgcgct tcgttagacg
     9901 gagagtagtt aacgaagtta cgccagcagt aaccagattc ctgttcggtc atgatctgga
     9961 tggtaacacc agaagagatc aggtcacgca gaccctgacg gttacgcggg tccgcgtggt
    10021 ggtacagacg cgcgatgtag atgaacaggg taacgtgcgg gtaacgagac aggaattcgg
    10081 tgatcgcacg agagcattca ccgcacggag accaagacag gaaccaggtg atagagcaac
    10141 gggtgttcgg gcagaagtaa cgttcggtgg tgaatttttc gatgaagtta acttcaacgt
    10201 gtttgttggt gttctgagag gtgtgacgcc agatagagtg acgaccaccc cagttgattt
    10261 cgtacagcag gcaggtttct ttacgcagtt cacgcgggtc gaagaaaact tcgaattcgt
    10321 gcggttcgat acgacgacgc agggtcgggt caaccgcaac cggaccggtt tcagaagact
    10381 tgtcatcgtc atccttgtaa tcgatatcat gatctttata atcaccgtca tggtctttgt
    10441 agtccatggt atatctcctt gtttcctgtg tgaaattgtt atccgctcac aattccacac
    10501 attatacgag ccgatgatta attgtcaaca gctcatttca gaatatttgc cagaaccgtt
    10561 atgatgtcgg cgcaaaaaac attatccaga acgggagtgc gccttgagcg acacgaatta
    10621 tgcagtgatt tacgacctgc acagccatac cacagcttcc gatggctgcc tgacgccaga
    10681 agcattggtg caccgtgcag tcgataagcc cggatcagct tgcaattcgc gcgcgaaggc
    10741 gaagcggcat ttacgttgac accatcgaat ggtgcaaaac ctttcgcggt atggcatgat
    10801 agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg
    10861 tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc
    10921 acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc
    10981 ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct
    11041 ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc
    11101 aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag
    11161 cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg
    11221 atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg
    11281 atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac
    11341 tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat
    11401 taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc
    11461 aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa
    11521 ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga
    11581 tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct
    11641 cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca
    11701 tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc
    11761 agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca
    11821 ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
    11881 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaaccccgg gctgcaggaa
    11941 ttcctcgaga ccggcgcgcc cctgtttgac agcttatcat cgcagatccg ggcaacgttg
    12001 ttgccattgg cggccgcaac gagctctagt gtacatcggg atcctggcag ttccctactc
    12061 tcgcatgggg agaccccaca ctaccatcgg cgctacggcg tttcacttct gagttcggca
    12121 tggggtcagg tgggaccacc gcgctactgc cgccaggcaa attctgtttt atcagaccgc
    12181 ttctgcgttc tgatttaatc tgtatcaggc tgaaaatctt ctctcatccg ccaaaacagc
    12241 caagcttatg gcttcgtttc gcagcgaact tggaaaaaaa atttttggcg attcgccgct
    12301 ttcgtataac gaagtaaaaa cctatctcag cgaacaccca ggcggaaccg cagaacgagc
    12361 ctggaatcat atttgtgcgc cttgaaatat ccctctccat accccataaa taaataatga
    12421 cattaatcct tcatgaagga gggcccttat ggagaaaaaa atcactggat ataccaccgt
    12481 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
    12541 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
    12601 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
    12661 ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
    12721 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
    12781 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
    12841 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt
    12901 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
    12961 caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
    13021 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
    13081 ctgcgatgag tggcagggcg gggcgtaaac caggtgctcg gcgtggcaag actcgcagcg
    13141 aacccgtaga aagccatgct ccagccgccc gcattggaga aattcttcaa attcccgttg
    13201 cacatagccc ggcaattcct ttccctgctc tgccataagc gcagcgaatg ccgggtaata
    13261 ctcgtcaacg atctgataga gaagggtttg ctcgggtcgg tggctctggt aacgaccagt
    13321 atcccgatcc cggctggccg tcctggccgc cacatgaggc atgttccgcg tccttgcaat
    13381 actgtgttta catacagtct atcgcttagc ggaaagttct tttaccctca gccgaaatgc
    13441 ctgccgttgc tagacattgc cagccagtgc ccgtcactcc cgtactaact gtcacgaacc
    13501 cctgcaataa ctgtcacgcc cccctgcaat aactgtcacg aacccctgca ataactgtca
    13561 cgcccccaaa cctgcaaacc cagcaggggc gggggctggc ggggtgttgg aaaaatccat
    13621 ccatgattat ctaagaataa tccactaggc gcggttatca gcgcccttgt ggggcgctgc
    13681 tgcccttgcc caatatgccc ggccagaggc cggatagctg gtctattcgc tgcgctaggc
    13741 tacacaccgc cccaccgctg cgcggcaggg ggaaaggcgg gcaaagcccg ctaaacccca
    13801 caccaaaccc cgcagaaata cgctggagcg cttttagccg ctttagcggc ctttccccct
    13861 acccgaaggg tgggggcgcg tgtgcagccc cgcagggcct gtctcggtcg atcattcagc
    13921 ccggctcatc cttctggcgt ggcggcagac cgaacaaggc gcggtcgtgg tcgcgttcaa
    13981 ggtacgcatc cattgccgcc atgagccgat cctccggcca ctcgctgctg ttcaccttgg
    14041 ccaaaatcat ggcccccacc agcaccttgc gccttgtttc gttcttgcgc tcttgctgct
    14101 gttcccttgc ccgcacccgc tgaatttcgg cattgattcg cgctcgttgt tcttcgagct
    14161 tggccagccg atccgccgcc ttgttgctcc ccttaaccat cttgacaccc cattgttaat
    14221 gtgctgtctc gtaggctatc atggaggcac agcggcggca atcccgaccc tactttgtag
    14281 gggagggcgc acttaccggt ttctcttcga gaaactggcc taacggccac ccttcgggcg
    14341 gtgcgctctc cgagggccat tgcatggagc cgaaaagcaa aagcaacagc gaggcagcat
    14401 ggcgatttat caccttacgg cgaaaaccgg cagcaggtcg ggcggccaat cggccagggc
    14461 caaggccgac tacatccagc gcgaaggcaa gtatgcccgc gacatggatg aagtcttgca
    14521 cgccgaatcc gggcacatgc cggagttcgt cgagcggccc gccgactact gggatgctgc
    14581 cgacctgtat gaacgcgcca atgggcggct gttcaaggag gtcgaatttg ccctgccggt
    14641 cgagctgacc ctcgaccagc agaaggcgct ggcgtccgag ttcgcccagc acctgaccgg
    14701 tgccgagcgc ctgccg
//
