LOCUS       sequence-344756-       11273 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For integration of J1-J23117-sfGFP, Sp.pCas9-dCas9, and 
            BBa_J23107-MCP-SoxS(R93A/S101A) with miniTn7T method.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11273)
  AUTHORS   Kiattisewee C, Dong C, Fontana J, Sugianto W, Peralta-Yahya P, 
            Carothers JM, Zalatan JG
  TITLE     Portable bacterial CRISPR transcriptional activation enables 
            metabolic engineering in Pseudomonas putida.
  JOURNAL   Metab Eng. 2021 Apr 27. pii: S1096-7176(21)00063-X. doi: 
            10.1016/j.ymben.2021.04.002.
  PUBMED    33930546
REFERENCE   2  (bases 1 to 11273)
  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: "Metab Eng. 
            2021 Apr 27. pii: S1096-7176(21)00063-X. doi: 
            10.1016/j.ymben.2021.04.002."
COMMENT     Sequence Label: pPPC002
FEATURES             Location/Qualifiers
     source          1..11273
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            55..163
                     /note="incP origin of transfer"
     rep_origin      complement(395..983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(475..494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1154..1945,1946..2014))
                     /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     1777..1796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2015..2119)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2187..2205
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2243..2265)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2365..2384
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2798..2814
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2799..2815
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      2843..2937
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      3040..3126
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    3172..3219
                     /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)."
     CDS             complement(3349..3882)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4071..4076,4077..4093,4094..4099))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    4140..4187
                     /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        4397..4431
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     RBS             4441..4452
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(4459..4461,4462..5175)
                     /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             4459..5172
                     /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(4489..4517)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5112..5132
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      5208..5279
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5295..5322
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        complement(5504..5582)
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     CDS             join(5878..5904,5905..5907,5908..8394,8395..8397,
                     8398..9984)
                     /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"
     terminator      10008..10079
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      10095..10122
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        10175..10209
                     /label=BBa_J23107
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.36"
     RBS             10219..10230
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     mobile_element  11004..11169
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 cactgacccg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagagc ttatcggcca
       61 gcctcgcaga gcaggattcc cgttgagcac cgccaggtgc gaataaggga cagtgaagaa
      121 ggaacacccg ctcgcgggtg ggcctacttc acctatcctg cccggctgac gccgttggat
      181 acaccaagga aagtctacac gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga
      241 tggatatacc gaaaaaatcg ctataatgac cccgaagcag ggttatgcag cggaaagtat
      301 accttaagga atcccctgat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      361 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      421 agcatcacaa aaatcgacgc tcgagtcaga ggtggcgaaa cccgacagga ctataaagat
      481 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      541 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
      601 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      661 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      721 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
      781 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
      841 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
      901 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
      961 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1021 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1081 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1141 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1201 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     1261 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     1321 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1381 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1441 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1501 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1561 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     1621 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     1681 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     1741 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     1801 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     1861 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     1921 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     1981 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     2041 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     2101 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     2161 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     2221 gtttcggtga cgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     2281 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2341 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     2401 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     2461 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     2521 agctagagga ccagccgcgt aacctggcaa aatcggttac ggttgagtaa taaatggatg
     2581 ccctgcgtaa gcaggtgtgg gcggacaata aagtcttaaa ctgaacaaaa tagatctaaa
     2641 ctatgacaat aaagtcttaa actagacaga atagttgtaa actgaaatca gtccagttat
     2701 gctgtgaaaa agcatactgg acttttgtta tggctaaagc aaactcttca ttttctgaag
     2761 tgcaaattgc ccgtcgtatt aaagaggggc gtggggttcg aggtcgacgg tatcgataag
     2821 ctagcttaat tagctgagct tggactcctg ttgatagatc cagtaatgac ctcagaactc
     2881 catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatcca
     2941 agctagactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt tattggtgcc
     3001 cttaaacgcc tggggtaatg actctctagc ttgaggcatc aaataaaacg aaaggctcag
     3061 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     3121 acaaatccgc cgctaggagc ttgcggcccg gacgatcgaa ttggggatct tgaagtacct
     3181 attccgaagt tcctattctc tagaaagtat aggaacttca gagcgctttt gaagctgatg
     3241 tgcttaaaaa cttactcaat ggaattctag atcgaattgg ccgcggcgtt gtgacaattt
     3301 accgaacaac tccgcggccg ggaagccgat ctcggcttga acgaattgtt aggtggcggt
     3361 acttgggtcg atatcaaagt gcatcacttc ttcccgtatg cccaactttg tatagagagc
     3421 cactgcggga tcgtcaccgt aatctgcttg cacgtagatc acataagcac caagcgcgtt
     3481 ggcctcatgc ttgaggagat tgatgagcgc ggtggcaatg ccctgcctcc ggtgctcgcc
     3541 ggagactgcg agatcataga tatagatctc actacgcggc tgctcaaact tgggcagaac
     3601 gtaagccgcg agagcgccaa caaccgcttc ttggtcgaag gcagcaagcg cgatgaatgt
     3661 cttactacgg agcaagttcc cgaggtaatc ggagtccggc tgatgttggg agtaggtggc
     3721 tacgtctccg aactcacgac cgaaaagatc aagagcagcc cgcatggatt tgacttggtc
     3781 agggccgagc ctacatgtgc gaatgatgcc catacttgag ccacctaact ttgttttagg
     3841 gcgactgccc tgctgcgtaa catcgttgct gctgcgtaac atcgttgctg ctccataaca
     3901 tcaaacatcg acccacggcg taacgcgctt gctgcttgga tgcccgaggc atagactgta
     3961 caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt
     4021 cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta cttgcattac agtttacgaa
     4081 ccgaacaggc ttatgtcaat tcgatccatt gctgttgaca aagggaatca ggggatcttg
     4141 aagttcctat tccgaagttc ctattctcta gaaagtatag gaacttcaga gcgcttttga
     4201 agctaattcg atcatgcatg ctgcaggcct acggtatcca ccggagacct atggcagcct
     4261 ccggccgcca taggacacct ttggttgcca agggtgacct atggtgacca tgggccacca
     4321 cgggcgacct caggtatcct gcggtgtcct gcggttacca aaggcgtcct ttgggttcca
     4381 ccggatacct ccggacttga cagctagctc agtcctaggg attgtgctag cgaattcatt
     4441 aaagaggaga aaggtaccat gagcaaagga gaagaacttt tcactggagt tgtcccaatt
     4501 cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtccgtgg agagggtgaa
     4561 ggtgatgcta caaacggaaa actcaccctt aaatttattt gcactactgg aaaactacct
     4621 gttccgtggc caacacttgt cactactctg acctatggtg ttcaatgctt ttcccgttat
     4681 ccggatcaca tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag
     4741 gaacgcacta tatctttcaa agatgacggg acctacaaga cgcgtgctga agtcaagttt
     4801 gaaggtgata cccttgttaa tcgtatcgag ttaaagggta ttgattttaa agaagatgga
     4861 aacattcttg gacacaaact cgagtacaac tttaactcac acaatgtata catcacggca
     4921 gacaaacaaa agaatggaat caaagctaac ttcaaaattc gccacaacgt tgaagatggt
     4981 tccgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt
     5041 ttaccagaca accattacct gtcgacacaa tctgtccttt cgaaagatcc caacgaaaag
     5101 cgtgaccaca tggtccttct tgagtttgta actgctgctg ggattacaca tggcatggat
     5161 gagctctaca aataaggatc caaactcgag taaggatctc caggcatcaa ataaaacgaa
     5221 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct
     5281 actagagtca cactggctca ccttcgggtg ggcctttctg cgtttataaa gcttttacga
     5341 aatcatcctg tggagcttag taggtttagc aagatggcag cgcctaaatg tagaatgata
     5401 aaaggattaa gagattaatt tccctaaaaa tgataaaaca agcgttttga aagcgcttgt
     5461 ttttttggtt tgcagtcaga gtagaataga agtatcaaaa aaagcaccga ctcggtgcca
     5521 ctttttcaag ttgataacgg actagcctta ttttaacttg ctatgctgtt ttgaatggtt
     5581 ccaacaagat tattttataa cttttataac aaataatcaa ggagaaattc aaagaaattt
     5641 atcagccata aaacaatact taatactata gaatgataac aaaataaact actttttaaa
     5701 agaattttgt gttataatct atttattatt aagtattggg taatattttt tgaagagata
     5761 ttttgaaaaa gaaaaattaa agcatattaa actaatttcg gaggtcatta aaactattat
     5821 tgaaatcatc aaactcatta tggatttaat ttaaactttt tattttagga ggcaaaaatg
     5881 gataagaaat actcaatagg cttagctatc ggcacaaata gcgtcggatg ggcggtgatc
     5941 actgatgaat ataaggttcc gtctaaaaag ttcaaggttc tgggaaatac agaccgccac
     6001 agtatcaaaa aaaatcttat aggggctctt ttatttgaca gtggagagac agcggaagcg
     6061 actcgtctca aacggacagc tcgtagaagg tatacacgtc ggaagaatcg tatttgttat
     6121 ctacaggaga ttttttcaaa tgagatggcg aaagtagatg atagtttctt tcatcgactt
     6181 gaagagtctt ttttggtgga agaagacaag aagcatgaac gtcatcctat ttttggaaat
     6241 atagtagatg aagttgctta tcatgagaaa tatccaacta tctatcatct gcgaaaaaaa
     6301 ttggtagatt ctactgataa agcggatttg cgcttaatct atttggcctt agcgcatatg
     6361 attaagtttc gtggtcattt tttgattgag ggagatttaa atcctgataa tagtgatgtg
     6421 gacaaactat ttatccagtt ggtacaaacc tacaatcaat tatttgaaga aaaccctatt
     6481 aacgcaagtg gagtagatgc taaagcgatt ctttctgcac gattgagtaa atcaagacga
     6541 ttagaaaatc tcattgctca gctccccggt gagaagaaaa atggcttatt tgggaatctc
     6601 attgctttgt cattgggttt gacccctaat tttaaatcaa attttgattt ggcagaagat
     6661 gctaaattac agctttcaaa agatacttac gatgatgatt tagataattt attggcgcaa
     6721 attggagatc aatatgctga tttgtttttg gcagctaaga atttatcaga tgctatttta
     6781 ctttcagata tcctaagagt aaatactgaa ataactaagg ctcccctatc agcttcaatg
     6841 attaaacgct acgatgaaca tcatcaagac ttgactcttt taaaagcttt agttcgacaa
     6901 caacttccag aaaagtataa agaaatcttt tttgatcaat caaaaaacgg atatgcaggt
     6961 tatattgatg ggggagctag ccaagaagaa ttttataaat ttatcaaacc aattttagaa
     7021 aaaatggatg gtactgagga attattggtg aaactaaatc gtgaagattt gctgcgcaag
     7081 caacggacct ttgacaacgg ctctattccc catcaaattc acttgggtga gctgcatgct
     7141 attttgagaa gacaagaaga cttttatcca tttttaaaag acaatcgtga gaagattgaa
     7201 aaaatcttga cttttcgaat tccttattat gttggtccat tggcgcgtgg caatagtcgt
     7261 tttgcatgga tgactcggaa gtctgaagaa acaattaccc catggaattt tgaagaagtt
     7321 gtcgataaag gtgcttcagc tcaatcattt attgaacgca tgacaaactt tgataaaaat
     7381 cttccaaatg aaaaagtact accaaaacat agtttgcttt atgagtattt tacggtttat
     7441 aacgaattga caaaggtcaa atatgttact gaaggaatgc gaaaaccagc atttctttca
     7501 ggtgaacaga agaaagccat tgttgattta ctcttcaaaa caaatcgaaa agtaaccgtt
     7561 aagcaattaa aagaagatta tttcaaaaaa atagaatgtt ttgatagtgt tgaaatttca
     7621 ggagttgaag atagatttaa tgcttcatta ggtacctacc atgatttgct aaaaattatt
     7681 aaagataaag attttttgga taatgaagaa aatgaagata tcttagagga tattgtttta
     7741 acattgacct tatttgaaga tagggagatg attgaggaaa gacttaaaac atatgctcac
     7801 ctctttgatg ataaggtgat gaaacagctt aaacgtcgcc gttatactgg ttggggacgt
     7861 ttgtctcgaa aattgattaa tggtattagg gataagcaat ctggcaaaac aatattagat
     7921 tttttgaaat cagatggttt tgccaatcgc aattttatgc agctgatcca tgatgatagt
     7981 ttgacattta aagaagacat tcaaaaagca caagtgtctg gacaaggcga tagtttacat
     8041 gaacatattg caaatttagc tggtagccct gctattaaaa aaggtatttt acagactgta
     8101 aaagttgttg atgaattggt caaagtaatg gggcggcata agccagaaaa tatcgttatt
     8161 gaaatggcac gtgaaaatca gacaactcaa aagggccaga aaaattcgcg agagcgtatg
     8221 aaacgaatcg aagaaggtat caaagaatta ggaagtcaga ttcttaaaga gcatcctgtt
     8281 gaaaatactc aattgcaaaa tgaaaagctc tatctctatt atctccaaaa tggaagagac
     8341 atgtatgtgg accaagaatt agatattaat cgtttaagtg attatgatgt cgatgccatt
     8401 gttccacaaa gtttccttaa agacgattca atagacaata aggtcttaac gcgttctgat
     8461 aaaaatcgtg gtaaatcgga taacgttcca agtgaagaag tagtcaaaaa gatgaaaaac
     8521 tattggagac aacttctaaa cgccaagtta atcactcaac gtaagtttga taatttaacg
     8581 aaagctgaac gtggaggttt gagtgaactt gataaagctg gttttatcaa acgccaattg
     8641 gttgaaactc gccaaatcac taagcatgtg gcacaaattt tggatagtcg catgaatact
     8701 aaatacgatg aaaatgataa acttattcga gaggttaaag tgattacctt aaaatctaaa
     8761 ttagtttctg acttccgaaa agatttccaa ttctataaag tacgtgagat taacaattac
     8821 catcatgccc atgatgcgta tctaaatgcc gtcgttggaa ctgctttgat taagaaatat
     8881 ccaaaacttg aatcggagtt tgtctatggt gattataaag tttatgatgt tcgtaaaatg
     8941 attgctaagt ctgagcaaga aataggcaaa gcaaccgcaa aatatttctt ttactctaat
     9001 atcatgaact tcttcaaaac agaaattaca cttgcaaatg gagagattcg caaacgccct
     9061 ctaatcgaaa ctaatgggga aactggagaa attgtctggg ataaagggcg agattttgcc
     9121 acagtgcgca aagtattgtc catgccccaa gtcaatattg tcaagaaaac agaagtacag
     9181 acaggcggat tctccaagga gtcaatttta ccaaaaagaa attcggacaa gcttattgct
     9241 cgtaaaaaag actgggatcc aaaaaaatat ggtggttttg atagtccaac ggtagcttat
     9301 tcagtcctag tggttgctaa ggtggaaaaa gggaaatcga agaagttaaa atccgttaaa
     9361 gagttactag ggatcacaat tatggaaaga agttcctttg aaaaaaatcc gattgacttt
     9421 ttagaagcta aaggatataa ggaagttaaa aaagacttaa tcattaaact acctaaatat
     9481 agtctttttg agttagaaaa cggtcgtaaa cggatgctgg ctagtgccgg agaattacaa
     9541 aaaggaaatg agctggctct gccaagcaaa tatgtgaatt ttttatattt agctagtcat
     9601 tatgaaaagt tgaagggtag tccagaagat aacgaacaaa aacaattgtt tgtggagcag
     9661 cataagcatt atttagatga gattattgag caaatcagtg aattttctaa gcgtgttatt
     9721 ttagcagatg ccaatttaga taaagttctt agtgcatata acaaacatag agacaaacca
     9781 atacgtgaac aagcagaaaa tattattcat ttatttacgt tgacgaatct tggagctccc
     9841 gctgctttta aatattttga tacaacaatt gatcgtaaac gatatacgtc tacaaaagaa
     9901 gttttagatg ccactcttat ccatcaatcc atcactggtc tttatgaaac acgcattgat
     9961 ttgagtcagc taggaggtga ctaactcgag taaggatctc caggcatcaa ataaaacgaa
    10021 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct
    10081 actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatacc tagagcgccc
    10141 ggaagagagt caattcaggg tggtgaatct gcagtttacg gctagctcag ccctaggtat
    10201 tatgctagcg aattcattaa agaggagaaa ggtaccatgg ggcccgcttc taactttact
    10261 cagttcgttc tcgtcgacaa tggcggaact ggcgacgtga ctgtcgcccc aagcaacttc
    10321 gctaacggga tcgctgaatg gatcagctct aactcgcgtt cacaggctta caaagtaacc
    10381 tgtagcgttc gtcagagctc tgcgcagaat cgcaaataca ccatcaaagt cgaggtgcct
    10441 aaaggcgcct ggcgttcgta cttaaatatg gaactaacca ttccaatttt cgccacgaat
    10501 tccgactgcg agcttattgt taaggcaatg caaggtctcc taaaagatgg aaacccgatt
    10561 ccctcagcaa tcgcagcaaa ctccggcatc tacggtggcg gaggtagcat gtcccatcag
    10621 aaaattattc aggatcttat cgcatggatt gacgagcata ttgaccagcc gcttaacatt
    10681 gatgtagtcg caaaaaaatc aggctattca aagtggtact tgcaacgaat gttccgcacg
    10741 gtgacgcatc agacgcttgg cgattacatt cgccaacgcc gcctgttact ggccgccgtt
    10801 gagttgcgca ccaccgagcg tccgattttt gatatcgcaa tggacctggg ttatgtctcg
    10861 cagcagacct tctcccgcgt tttcgcgcgg cagtttgatc gcactcccgc ggattatcgc
    10921 caccgcctgt aagcggccgc cacgcaaaaa accccgcttc ggcggggttt tttcgcaagc
    10981 ttctcgcgaa ggccttgcag gccaaccaga taagtgaaat ctagttccaa actattttgt
    11041 catttttaat tttcgtatta gcttacgacg ctacacccag ttcccatcta ttttgtcact
    11101 cttccctaaa taatccttaa aaactccatt tccacccctc ccagttccca actattttgt
    11161 ccgcccacag cggggcattt ttcttcctgt tatgtttggg cgctgcatta atgaatcggc
    11221 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gct
//
