LOCUS       sequence-326621-       11278 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Escherichia coli – Staphylococcus aureus shuttle vector for plasmid 
            curing in Gram-positive bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11278)
  AUTHORS   Dorado-Morales P, Garcillan-Barcia MP, Lasa I, Solano C
  TITLE     Fitness Cost Evolution of Natural Plasmids of Staphylococcus aureus.
  JOURNAL   mBio. 2021 Feb 23;12(1). pii: mBio.03094-20. doi: 
            10.1128/mBio.03094-20.
  PUBMED    33622733
REFERENCE   2  (bases 1 to 11278)
  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: "mBio."; 
            date: "2021-02-23"; volume: "12(1). pii: mBio.03094-20. doi"; pages:
            " 10.1128/mBio.03094-20"
COMMENT     Sequence Label: pEMPTY::sgRNA2
FEATURES             Location/Qualifiers
     source          1..11278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1176..1198
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1236..1254)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1322..1426
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1427..1495,1496..2287)
                     /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(1645..1664)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2458..3046
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2947..2966
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_RNA        3260..3335
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      complement(3416..3463)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3459..3477
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3512..7618)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(3515..7618)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     protein_bind    7661..7679
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             7786..8412
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(7862..7881)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(8493..8510)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8493..8509)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8502..8524)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ataggcttgg ttatgccggt actgccgggc ctcttgcggg atcgaggggg cccacctagg
       61 tattatcaag ataagaaaga aaaggatttt tcgctacgct caaatccttt aaaaaaacac
      121 aaaagaccac attttttaat gtggtctttt attcttcaac taaagcaccc attagttcaa
      181 caaacgaaaa ttggataaag tgggatattt ttaaaatata tatttatgtt acagtaatat
      241 tgacttttaa aaaaggattg attctaatga agaaagcaga caagtaagcc tcctaaattc
      301 actttagata aaaatttagg aggcatatca aatgaacttt aataaaattg atttagacaa
      361 ttggaagaga aaagagatat ttaatcatta tttgaaccaa caaacgactt ttagtataac
      421 cacagaaatt gatattagtg ttttataccg aaacataaaa caagaaggat ataaatttta
      481 ccctgcattt attttcttag tgacaagggt gataaactca aatacagctt ttagaactgg
      541 ttacaatagc gacggagagt taggttattg ggataagtta gagccacttt atacaatttt
      601 tgatggtgta tctaaaacat tctctggtat ttggactcct gtaaagaatg acttcaaaga
      661 gttttatgat ttataccttt ctgatgtaga gaaatataat ggttcgggga aattgtttcc
      721 caaaacacct atacctgaaa atgctttttc tctttctatt attccatgga cttcatttac
      781 tgggtttaac ttaaatatca ataataatag taattacctt ctacccatta ttacagcagg
      841 aaaattcatt aataaaggta attcaatata tttaccgcta tctttacagg tacatcattc
      901 tgtttgtgat ggttatcatg caggattgtt tatgaactct attcaggaat tgtcagatag
      961 gcctaatgac tggcttttat aatatgagat aatgccgact gtacttttta cagtcggttt
     1021 tctaatgtca ctaacctgcc ccgttagttg aagaaggccg cggcctcgag cggccgcata
     1081 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     1141 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     1201 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     1261 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     1321 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1381 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1441 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1501 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1561 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1621 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     1681 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     1741 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     1801 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     1861 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     1921 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     1981 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2041 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2101 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     2161 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     2221 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2281 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     2341 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     2401 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2461 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2521 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2581 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     2641 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2701 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2761 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2821 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2881 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2941 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3001 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3061 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     3121 atcccctgat tctgtggata accgtattac cgcctttgag tgagctggcg gccgctgcat
     3181 gcctgcaggg cgcgccttga caaattgcag taggcatgac aaaatggact cacaagtttt
     3241 gggattgtta agggttccgg ttttagagct agaaatagca agttaaaata aggctagtcc
     3301 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttcgcaaaaa accccgcttc
     3361 ggcggggttt tttcgctagt cggtgtgcgc aaccttaagc ccgggtacca gatctcaaaa
     3421 aacccctcaa gacccgttta gaggccccaa ggggttatgc tagttattgc tcagcggtgg
     3481 cagcagccta ggataccggt aacatgggat ctcagtcacc tcctagctga ctcaaatcaa
     3541 tgcgtgtttc ataaagacca gtgatggatt gatggataag agtggcatct aaaacttctt
     3601 ttgtagacgt atatcgttta cgatcaattg ttgtatcaaa atatttaaaa gcagcgggag
     3661 ctccaagatt cgtcaacgta aataaatgaa taatattttc tgcttgttca cgtattggtt
     3721 tgtctctatg tttgttatat gcactaagaa ctttatctaa attggcatct gctaaaataa
     3781 cacgcttaga aaattcactg atttgctcaa taatctcatc taaataatgc ttatgctgct
     3841 ccacaaacaa ttgtttttgt tcgttatctt ctggactacc cttcaacttt tcataatgac
     3901 tagctaaata taaaaaattc acatatttgc ttggcagagc cagctcattt cctttttgta
     3961 attctccggc actagccagc atccgtttac gaccgttttc taactcaaaa agactatatt
     4021 taggtagttt aatgattaag tcttttttaa cttccttata tcctttagct tctaaaaagt
     4081 caatcggatt tttttcaaag gaacttcttt ccataattgt gatccctagt aactctttaa
     4141 cggattttaa cttcttcgat ttcccttttt ccaccttagc aaccactagg actgaataag
     4201 ctaccgttgg actatcaaaa ccaccatatt tttttggatc ccagtctttt ttacgagcaa
     4261 taagcttgtc cgaatttctt tttggtaaaa ttgactcctt ggagaatccg cctgtctgta
     4321 cttctgtttt cttgacaata ttgacttggg gcatggacaa tactttgcgc actgtggcaa
     4381 aatctcgccc tttatcccag acaatttctc cagtttcccc attagtttcg attagagggc
     4441 gtttgcgaat ctctccattt gcaagtgtaa tttctgtttt gaagaagttc atgatattag
     4501 agtaaaagaa atattttgcg gttgctttgc ctatttcttg ctcagactta gcaatcattt
     4561 tacgaacatc ataaacttta taatcaccat agacaaactc cgattcaagt tttggatatt
     4621 tcttaatcaa agcagttcca acgacggcat ttagatacgc atcatgggca tgatggtaat
     4681 tgttaatctc acgtacttta tagaattgga aatcttttcg gaagtcagaa actaatttag
     4741 attttaaggt aatcacttta acctctcgaa taagtttatc attttcatcg tatttagtat
     4801 tcatgcgact atccaaaatt tgtgccacat gcttagtgat ttggcgagtt tcaaccaatt
     4861 ggcgtttgat aaaaccagct ttatcaagtt cactcaaacc tccacgttca gctttcgtta
     4921 aattatcaaa cttacgttga gtgattaact tggcgtttag aagttgtctc caatagtttt
     4981 tcatcttttt gactacttct tcacttggaa cgttatccga tttaccacga tttttatcag
     5041 aacgcgttaa gaccttattg tctattgaat cgtctttaag gaaactttgt ggaacaatgt
     5101 gatcgacatc ataatcactt aaacgattaa tatctaattc ttggtccaca tacatgtctc
     5161 ttccattttg gagataatag agatagagct tttcattttg caattgagta ttttcaacag
     5221 gatgctcttt aagaatctga cttcctaatt ctttgatacc ttcttcgatt cgtttcatac
     5281 gctctcgcga atttttctgg cccttttgag ttgtctgatt ttcacgtgcc atttcaataa
     5341 cgatattttc tggcttatgc cgccccatta ctttgaccaa ttcatcaaca acttttacag
     5401 tctgtaaaat acctttttta atagcagggc taccagctaa atttgcaata tgttcatgta
     5461 aactatcgcc ttgtccagac acttgtgctt tttgaatgtc ttctttaaat gtcaaactat
     5521 catcatggat cagctgcata aaattgcgat tggcaaaacc atctgatttc aaaaaatcta
     5581 atattgtttt gccagattgc ttatccctaa taccattaat caattttcga gacaaacgtc
     5641 cccaaccagt ataacggcga cgtttaagct gtttcatcac cttatcatca aagaggtgag
     5701 catatgtttt aagtctttcc tcaatcatct ccctatcttc aaataaggtc aatgttaaaa
     5761 caatatcctc taagatatct tcattttctt cattatccaa aaaatcttta tctttaataa
     5821 tttttagcaa atcatggtag gtacctaatg aagcattaaa tctatcttca actcctgaaa
     5881 tttcaacact atcaaaacat tctatttttt tgaaataatc ttcttttaat tgcttaacgg
     5941 ttacttttcg atttgttttg aagagtaaat caacaatggc tttcttctgt tcacctgaaa
     6001 gaaatgctgg ttttcgcatt ccttcagtaa catatttgac ctttgtcaat tcgttataaa
     6061 ccgtaaaata ctcataaagc aaactatgtt ttggtagtac tttttcattt ggaagatttt
     6121 tatcaaagtt tgtcatgcgt tcaataaatg attgagctga agcaccttta tcgacaactt
     6181 cttcaaaatt ccatggggta attgtttctt cagacttccg agtcatccat gcaaaacgac
     6241 tattgccacg cgccaatgga ccaacataat aaggaattcg aaaagtcaag attttttcaa
     6301 tcttctcacg attgtctttt aaaaatggat aaaagtcttc ttgtcttctc aaaatagcat
     6361 gcagctcacc caagtgaatt tgatggggaa tagagccgtt gtcaaaggtc cgttgcttgc
     6421 gcagcaaatc ttcacgattt agtttcacca ataattcctc agtaccatcc attttttcta
     6481 aaattggttt gataaattta taaaattctt cttggctagc tcccccatca atataacctg
     6541 catatccgtt ttttgattga tcaaaaaaga tttctttata cttttctgga agttgttgtc
     6601 gaactaaagc ttttaaaaga gtcaagtctt gatgatgttc atcgtagcgt ttaatcattg
     6661 aagctgatag gggagcctta gttatttcag tatttactct taggatatct gaaagtaaaa
     6721 tagcatctga taaattctta gctgccaaaa acaaatcagc atattgatct ccaatttgcg
     6781 ccaataaatt atctaaatca tcatcgtaag tatcttttga aagctgtaat ttagcatctt
     6841 ctgccaaatc aaaatttgat ttaaaattag gggtcaaacc caatgacaaa gcaatgagat
     6901 tcccaaataa gccatttttc ttctcaccgg ggagctgagc aatgagattt tctaatcgtc
     6961 ttgatttact caatcgtgca gaaagaatcg ctttagcatc tactccactt gcgttaatag
     7021 ggttttcttc aaataattga ttgtaggttt gtaccaactg gataaatagt ttgtccacat
     7081 cactattatc aggatttaaa tctccctcaa tcaaaaaatg accacgaaac ttaatcatat
     7141 gcgctaaggc caaatagatt aagcgcaaat ccgctttatc agtagaatct accaattttt
     7201 ttcgcagatg atagatagtt ggatatttct catgataagc aacttcatct actatatttc
     7261 caaaaatagg atgacgttca tgcttcttgt cttcttccac caaaaaagac tcttcaagtc
     7321 gatgaaagaa actatcatct actttcgcca tctcatttga aaaaatctcc tgtagataac
     7381 aaatacgatt cttccgacgt gtataccttc tacgagctgt ccgtttgaga cgagtcgctt
     7441 ccgctgtctc tccactgtca aataaaagag cccctataag attttttttg atactgtggc
     7501 ggtctgtatt tcccagaacc ttgaactttt tagacggaac cttatattca tcagtgatca
     7561 ccgcccatcc gacgctattt gtgccgatat ctaagcctat tgagtatttc ttatccataa
     7621 ataatcatcc tcctaaagat ctcaagctta ttttaattat actctatcaa tgatagagtg
     7681 tcaatatttt ttttagtttt tcatgaactc gaggggatcc aaataaaaaa ctagtttgac
     7741 aaataactct atcaatgata taatgtcaac aaaaaggagg aattaatgat gtctagatta
     7801 gataaaagta aagtgattaa cagcgcatta gagctgctta atgaggtcgg aatcgaaggt
     7861 ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc agcctacatt gtattggcat
     7921 gtaaaaaata agcgggcttt gctcgacgcc ttagccattg agatgttaga taggcaccat
     7981 actcactttt gccctttaga aggggaaagc tggcaagatt ttttacgtaa taacgctaaa
     8041 agttttagat gtgctttact aagtcatcgc gatggagcaa aagtacattt aggtacacgg
     8101 cctacagaaa aacagtatga aactctcgaa aatcaattag cctttttatg ccaacaaggt
     8161 ttttcactag agaatgcatt atatgcactc agcgctgtgg ggcattttac tttaggttgc
     8221 gtattggaag atcaagagca tcaagtcgct aaagaagaaa gggaaacacc tactactgat
     8281 agtatgccgc cattattacg acaagctatc gaattatttg atcaccaagg tgcagagcca
     8341 gccttcttat tcggccttga attgatcata tgcggattag aaaaacaact taaatgtgaa
     8401 agtgggtctt aaaagcagca taaccttttt ccgtgatggt aacttcacgg taaccaagat
     8461 gtcgagttat cgataccgtc gacctggaat tcactggccg tcgttttaca acgtcgtgac
     8521 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     8581 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     8641 ggcgaatggc gcctgagggt tgccagagtt aaaggatcaa gtaatacaaa cgggatacaa
     8701 agacataatc aaagagagaa taaaaactat aataataaag acataaatca tgaggaaaca
     8761 tataaaaatt atgatttgat taacgcacaa aatataaagt ataaagataa aattgatgaa
     8821 acgattgatg agaattattc agggaaacgt aaaattcggt cagatgcaat tcgacatgtg
     8881 gacggactgg ttacaagtga taaagatttc tttgatgatt taagcggaga agaaatagaa
     8941 cgatttttta aagatagctt ggagtttcta gaaaatgaat acggtaagga aaatatgctg
     9001 tatgcgactg tccatctgga tgaaagagtc ccacatatgc actttggttt tgtcccttta
     9061 acagaggacg ggagattgtc tgcaaaagaa cagttaggca acaagaaaga ctttactcaa
     9121 ttacaagata gatttaatga gtatgtgaat gagaaaggtt atgaacttga aagaggcacg
     9181 tccaaagagg ttacagaacg agaacataaa gcgatggatc agtacaagaa agatactgta
     9241 tttcataaac aggaactgca agaagttaag gatgagttac agaaggcaaa taagcagtta
     9301 cagagtggaa tagagcatat gaggtctacg aaaccctttg attatgaaaa tgagcgtaca
     9361 ggtttgttct ctggacgtga agagactggt agaaagatat taactgctga tgaatttgaa
     9421 cgcctgcaag aaacaatctc ttctgcagaa cggattgttg atgattacga aaatattaag
     9481 agcacagact attacacaga aaatcaagaa ttaaaaaaac gtagagagag tttgaaagaa
     9541 gtagtgaata catggaaaga ggggtatcac gaaaaaagta aagaggttaa taaattaaag
     9601 cgagagaatg atagtttgaa tgagcagttg aatgtatcag agaaatttca agatagtaca
     9661 gtgactttat atcgtgctgc gagggcgaat ttccctgggt ttgagaaagg gtttaatagg
     9721 cttaaagaga aattctttaa tgattccaaa ttcgagcgtg tgggacagtt tatggatgtt
     9781 gtacaggata atgtccagaa ggtcgataga aagcgtgaga aacagcgtac agacgattta
     9841 gagatgtaga ggtactttta tgccgagaaa actttttgcg tgtgacagtc cttaaaatat
     9901 acttagagcg taagcgaaag tagtagcgac agctattaac tttcggttgc aaagctctag
     9961 gatttttaat ggacgcagcg catcacacgc aaaaaggaaa ttggaataaa tgcgaaattt
    10021 gagatgttaa ttaaagacct ttttgaggtc tttttttctt agatttttgg ggttatttag
    10081 gggagaaaac ataggggggt actacgacct cccccctagg tgtccattgt ccattgtcca
    10141 aacaaataaa taaatattgg gtttttaatg ttaaaaggtt gttttttatg ttaaagtgaa
    10201 aaaaacagat gttgggaggt acagtgatgg ttgtagatag aaaagaagag aaaaaagttg
    10261 ctgttacttt aagacttaca acagaagaaa atgagatatt aaatagaatc aaagaaaaat
    10321 ataatattag caaatcagat gcaaccggta ttctaataaa aaaatatgca aaggaggaat
    10381 acggtgcatt ttaaacaaaa aaagatagac agcactggca tgctgcctat ctatgactaa
    10441 attttgttaa atgtattagc accgttatta tatcatgagc gaaaatgtaa taaaagaaac
    10501 tgaaaacaag aaaaattcaa gaggacgtaa ttggacattt gttttatatc cagaatcagc
    10561 aaaagccgag tggttagagt atttaaaaga gttacacatt caatttgtag tgtctccatt
    10621 acatgatagg gatactgata cagaagatag gatgaaaaaa gagcattatc atattctagt
    10681 gatgtatgag ggtaataaat cttatgaaca gataaaaata attacagaag aattgaatgc
    10741 gactattccg cagattgcag gaagtgtgaa aggtcttgtg agatatatgc ttcacatgga
    10801 cgatcctaat aaatttaaat atcaaaaaga agatatgata gtttatggcg gtgtagatgt
    10861 tgatgaatta ttaaagaaaa caacaacaga tagatataaa ttaattaaag aaatgattga
    10921 gtttattgat gaacaaggaa tcgtagaatt taagagttta atggattatg caatgaagtt
    10981 taaatttgat gattggttcc cgcttttatg tgataactcg gcgtatgtta ttcaagaata
    11041 tataaaatca aatcggtata aatctgaccg atagattttg aatttaagag tgtcacaaga
    11101 cactcttttt tcgcaccaac gaaaactggt ttaagccgac tgcgcaaaag acataatcga
    11161 taagctttaa tgcggtagtt tatcacagtt aaattgctaa cgcagtcagg caccgtgtat
    11221 gaaatctaac aatgcgctca tcgtcatcct cggcaccgtc accctggatg ctgtaggc
//
