LOCUS       sequence-321808-       11648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BAC-based, lacUV5 regulon and Cre recombinase in loxP and lox5171 
            sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11648)
  AUTHORS   Wang G, Zhao Z, Ke J, Engel Y, Shi YM, Robinson D, Bingol K, Zhang 
            Z, Bowen B, Louie K, Wang B, Evans R, Miyamoto Y, Cheng K, Kosina S,
            De Raad M, Silva L, Luhrs A, Lubbe A, Hoyt DW, Francavilla C, Otani 
            H, Deutsch S, Washton NM, Rubin EM, Mouncey NJ, Visel A, Northen T, 
            Cheng JF, Bode HB, Yoshikuni Y
  TITLE     CRAGE enables rapid activation of biosynthetic gene clusters in 
            undomesticated bacteria.
  JOURNAL   Nat Microbiol. 2019 Dec;4(12):2498-2510. doi: 
            10.1038/s41564-019-0573-8. Epub 2019 Oct 14.
  PUBMED    31611640
REFERENCE   2  (bases 1 to 11648)
  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"; doi: 
            "10.1038/s41564-019-0573-8"; journalName: "Nat Microbiol"; date: 
            "2019-12"; volume: "4"; issue: "12"; pages: "2498-2510"
COMMENT     Sequence Label: pW5Y_Apr
FEATURES             Location/Qualifiers
     source          1..11648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..24,25..31)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    39..55
                     /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        join(96..101,102..119,120..126)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    134..150
                     /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)."
     CDS             199..1230
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTTGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     oriT            1460..1568
                     /note="incP origin of transfer"
     primer_bind     1613..1631
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1656..1673)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1656..1672)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1665..1687)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    1810..2283
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     CDS             complement(2356..3327)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(3327..4502)
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTVSMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     promoter        4716..4931
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4932..5735
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     misc_feature    5899..6403
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     misc_feature    6618..6868
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(6871..7626)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      7717..7936
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    complement(8865..8898)
                     /label=lox5171
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTGTAC) (Shaw et al., 2021). lox5171 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     promoter        8896..8998
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     misc_feature    9906..10013
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     9922..9938
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     9923..9939
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9973..9989)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9973..9989)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    10040..10073
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(10282..11364)
                     /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     11326..11345
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(11365..11442)
                     /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."
     primer_bind     11570..11592
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tttacacttt atgcttccgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
       61 cacaggaaac agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
      121 ataatgtgtg gaattgtgag cggataacaa tttcacacag gaaacaggcc tcgactctag
      181 actgcaggta ccgttgacat gtccaattta ctgaccgtac accaaaattt gcctgcatta
      241 ccggtcgatg caacgagtga tgaggttcgc aagaacctga tggacatgtt cagggatcgc
      301 caggcgtttt ctgagcatac ctggaaaatg cttctgtccg tttgccggtc gtgggcggca
      361 tggtgcaagt tgaataaccg gaaatggttt cccgcagaac ctgaagatgt tcgcgattat
      421 cttctatatc ttcaggcgcg cggtctggca gtaaaaacta tccagcaaca tttgggccag
      481 ctaaacatgc ttcatcgtcg gtccgggctg ccacgaccaa gtgacagcaa tgctgtttca
      541 ctggttatgc ggcggatccg aaaagaaaac gttgatgccg gtgaacgtgc aaaacaggct
      601 ctagcgttcg aacgcactga tttcgaccag gttcgttcac tcatggaaaa tagcgatcgc
      661 tgccaggata tacgtaatct ggcatttctg gggattgctt ataacaccct gttacgtata
      721 gccgaaattg ccaggatcag ggttaaagat atctcacgta ctgacggtgg gagaatgtta
      781 atccatattg gcagaacgaa aacgctggtt agcaccacag gtgtagagaa ggcacttagc
      841 ctgggggtaa ctaaactggt cgagcgatgg atttccgtct ctggtgtagc tgatgatccg
      901 aataactacc tgttttgccg ggtcagaaaa aatggtgttg ccgcgccatc tgccaccagc
      961 cagctatcaa ctcgcgccct ggaagggatt tttgaagcaa ctcatcgatt gatttacggc
     1021 gctaaggatg actctggtca gagatacctg gcctggtctg gacacagtgc ccgtgtcgga
     1081 gccgcgcgag atatggcccg cgctggagtt tcaataccgg agatcatgca agctggtggc
     1141 tggaccaatg taaatattgt catgaactat atccgtaacc tggatagtga aacaggggca
     1201 atggtgcgcc tgctggaaga tggcgattag taagaattcc cggggatccg gtgattgatt
     1261 gagcaagctt ggcactggct gatcagctag cccatgggta tggacagttt tccctttgat
     1321 atgtaacggt cgctgcataa ccctgcttcg gggtcattat agcgattttt tcggtatatc
     1381 catccttttt cgcacgatat acaggatttt gccaaagggt tcgtgtagac tttccttggt
     1441 gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt
     1501 ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag
     1561 gctggccgat aagctctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     1621 tatcacgagg ccctttcgtc ttcaagaatt aattcactgg ccgtcgtttt acaacgtcgt
     1681 gactgggaaa accctggcgt tacccaacac ataaccgagg ttactccgtt ctacaggtta
     1741 cgacgacatg tcaatacttg cccttgacag gcattgatgg aatcgtagtc tcacgctgat
     1801 agtctgatcg acaatacaag tgggaccgtg gtcccagacc gataatcaga ccgacaacac
     1861 gagtgggatc gtggtcccag actaataatc agaccgacga tacgagtggg accgtggtcc
     1921 cagactaata atcagaccga cgatacgagt gggaccgtgg ttccagacta ataatcagac
     1981 cgacgatacg agtgggaccg tggtcccaga ctaataatca gaccgacgat acgagtggga
     2041 ccatggtccc agactaataa tcagaccgac gatacgagtg ggaccgtggt cccagtctga
     2101 ttatcagacc gacgatacga gtgggaccgt ggtcccagac taataatcag accgacgata
     2161 cgagtgggac cgtggtccca gactaataat cagaccgacg atacgagtgg gaccgtggtc
     2221 ccagtctgat tatcagaccg acgatacaag tggaacagtg ggcccagaga gaatattcag
     2281 gccagttatg ctttctggcc tgtaacaaag gacattaagt aaagacagat aaacgtagac
     2341 taaaacgtgg tcgcatcagg gtgctggctt ttcaagttcc ttaagaatgg cctcaatttt
     2401 ctctatacac tcagttggaa cacgagacct gtccaggtta agcaccattt tatcgccctt
     2461 atacaatact gtcgctccag gagcaaactg atgtcgtgag cttaaactag ttcttgatgc
     2521 agatgacgtt ttaagcacag aagttaaaag agtgataact tcttcagctt caaatatcac
     2581 cccagctttt ttctgctcat gaaggttaga tgcctgctgc ttaagtaatt cctctttatc
     2641 tgtaaaggct ttttgaagtg catcacctga ccgggcagat agttcaccgg ggtgagaaaa
     2701 aagagcaaca actgatttag gcaatttggc ggtgttgata cagcgggtaa taatcttacg
     2761 tgaaatattt tccgcatcag ccagcgcaga aatatttcca gcaaattcat tctgcaatcg
     2821 gcttgcataa cgctgaccac gttcataagc acttgttggg cgataatcgt tacccaatct
     2881 ggataatgca gccatctgct catcatccag ctcgccaacc agaacacgat aatcactttc
     2941 ggtaagtgca gcagctttac gacggcgact cccatcggca atttctatga caccagatac
     3001 tcttcgaccg aacgccggtg tctgttgacc agtcagtaga aaagaaggga tgagatcatc
     3061 cagtgcgtcc tcagtaagca gctcctggtc acgttcatta cctgaccata cccgagaggt
     3121 cttctcaaca ctatcacccc ggagcacttc aagagtaaac ttcacatccc gaccacatac
     3181 aggcaaagta atggcattac cgcgagccat tactcctacg cgcgcaatta acgaatccac
     3241 catcggggca gctggtgtcg ataacgaagt atcttcaacc ggttgagtat tgagcgtatg
     3301 ttttggaata acaggcgcac gcttcattat ctaatctccc agcgtggttt aatcagacga
     3361 tcgaaaattt cattgcagac aggttcccaa atagaaagag catttctcca ggcaccagtt
     3421 gaagagcgtt gatcaatggc ctgttcaaaa acagttctca tccggatctg acctttacca
     3481 acttcatccg tttcacgtac aacatttttt agaaccatgc ttccccaggc atcccgaatt
     3541 tgctcctcca tccacgggga ctgagagcca ttactattgc tgtatttggt aagcaaaata
     3601 cgtacatcag gctcgaaccc tttaagatca acgttcttga gcagatcacg aagcatatcg
     3661 aaaaactgca gtgcggaggt gtagtcaaac aactcagcag gcgtgggaac aatcagcaca
     3721 tcagcagcac atacgacatt aatcgtgccg atacccaggt taggcgcgct gtcaataact
     3781 atgacatcat agtcatgagc aacagtttca atggccagtc ggagcatcag gtgtggatcg
     3841 gtgggcagtt taccttcatc aaatttgccc attaactcag tttcaatacg gtgcagagcc
     3901 agacaggaag gaataatgtc aagccccggc cagcaagtgg gctttattgc ataagtgaca
     3961 tcgtcctttt ccccaagata gaaaggcagg agagtgtctt ctgcatgaat atgaagatct
     4021 ggtacccatc cgtgatacat tgagactgtt ccctgggggt cgttaccttc cacgagcaaa
     4081 acacgtagcc ccttcagagc cagatcctga gcaagatgaa cagaaactga ggttttgtaa
     4141 acgccacctt tatgggcagc aaccccgatc accggtggaa atacgtcttc agcacgtcgc
     4201 aatcgcgtac caaacacatc acgcatatga ttaatttgtt caattgtata accaacacgt
     4261 tgctcaaccc gtcctcgaat ttccatatcc gggtgcggta gtcgccctgc tttctcggca
     4321 tctctgatag cctgagaaga aaccccaact aaatccgctg cttcacctat tctccagcgc
     4381 cgggttattt tcctcgcttc cgggctgtca tcattaaact gtgcaatggc gatagccttc
     4441 gtcatttcat gaccagcgtt tatgcactgg ttaagtgttt ccatgagttt cattctgaac
     4501 atcctttaat cattgctttg cgttttttta ttaaatcttg caatttactg caaagcaaca
     4561 acaaaatcgc aaagtcatca aaaaaccgca aagttgttta aaataagagc aacactacaa
     4621 aaggagataa gaagagcaca tacctcagtc acttattatc actagcgagt gcaccatatt
     4681 taaatgcccg ggcttaatta aggcgcgccc agcttttcaa ttcatcattt tttttttatt
     4741 cttttttttg atttcggttt ccttgaaatt tttttgattc ggtaatctcc gaacagaagg
     4801 aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt agtgttgaag
     4861 aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacc tgcaggaaac
     4921 gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg
     4981 ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg
     5041 atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt
     5101 tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc
     5161 taaaggcatt atccgccaag tacaattttt tactcttcga agacagaaaa tttgctgaca
     5221 ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag
     5281 acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg
     5341 cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg
     5401 gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag
     5461 attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt
     5521 ggttgattat gacacccggt gtgggtttag atgacaaggg agacgcattg ggtcaacagt
     5581 atagaaccgt ggatgatgtg gtctctacag gatctgacat tattattgtt ggaagaggac
     5641 tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg
     5701 aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat
     5761 gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta cccgggaatc
     5821 tcggtcgtaa tgatttttat aatgacgaaa aaaaaaaaat tggaaagaaa aagctgggcg
     5881 cgccggccgg cccttttcat cacgtgctat aaaaataatt ataatttaaa ttttttaata
     5941 taaatatata aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt
     6001 tttccgaaga tgtaaaagac tctaggggga tcgccaacaa atactacctt ttatcttgct
     6061 cttcctgctc tcaggtatta atgccgaatt gtttcatctt gtctgtgtag aagaccacac
     6121 acgaaaatcc tgtgatttta cattttactt atcgttaatc gaatgtatat ctatttaatc
     6181 tgcttttctt gtctaataaa tatatatgta aagtacgctt tttgttgaaa ttttttaaac
     6241 ctttgtttat tttttttttc ttcattccgt aactcttcta ccttctttat ttactttcta
     6301 aaatccaaat acaaaacata aaaataaata aacacagagt aaattcccaa attattccat
     6361 cattaaaaga tacgaggcgc gtgtaagtta caggcaagcg atcggccggc ccgggcattt
     6421 aaatgcaggc cgcgtacgcg tcgacgcgta ccgaatgctc gccgcagccg tgtaaccgag
     6481 catagcgagc gaactggcga ggaagcaaag aagaactgtt ctgtcagata gctcttacgc
     6541 tcagcgcaag aagaaatatc caccgtggga aaaactccag gtagaggtac acacgcggat
     6601 agccaattca gagtaataaa ctgtgataat caaccctcat caatgatgac gaactaaccc
     6661 ccgatatcag gtcacatgac gaagggaaag agaaggaaat caactgtgac aaactgccct
     6721 caaatttggc ttccttaaaa attacagttc aaaaagtatg agaaaatcca tgcaggctga
     6781 aggaaacagc aaaactgtga caaattaccc tcagtaggtc agaacaaatg tgacgaacca
     6841 ccctcaaatc tgtgacagat aaccctcaga ctatcctgtc gtcatggaag tgatatcgcg
     6901 gaaggaaaat acgatatgag tcgtctggcg gcctttcttt ttctcaatgt atgagaggcg
     6961 cattggagtt ctgctgttga tctcattaac acagacctgc aggaagcggc ggcggaagtc
     7021 aggcatacgc tggtaacttt gaggcagctg gtaacgctct atgatccagt cgattttcag
     7081 agagacgatg cctgagccat ccggcttacg atactgacac agggattcgt ataaacgcat
     7141 ggcatacgga ttggtgattt cttttgtttc actaagccga aactgcgtaa accggttctg
     7201 taacccgata aagaagggaa tgagatatgg gttgatatgt acactgtaaa gccctctgga
     7261 tggactgtgc gcacgtttga taaaccaagg aaaagattca tagccttttt catcgccggc
     7321 atcctcttca gggcgataaa aaaccacttc cttccccgcg aaactcttca atgcctgccg
     7381 tatatcctta ctggcttccg cagaggtcaa tccgaatatt tcagcatatt tagcaacatg
     7441 gatctcgcag ataccgtcat gttcctgtag ggtgccatca gattttctga tctggtcaac
     7501 gaacagatac agcatacgtt tttgatcccg ggagagacta tatgccgcct cagtgaggtc
     7561 gtttgactgg acgattcgcg ggctattttt acgtttcttg tgattgataa ccgctgtttc
     7621 cgccatgaca gatccatgtg aagtgtgaca agtttttaga ttgtcacact aaataaaaaa
     7681 gagtcaataa gcagggataa ctttgtgaaa aaacagcttc ttctgagggc aatttgtcac
     7741 agggttaagg gcaatttgtc acagacagga ctgtcatttg agggtgattt gtcacactga
     7801 aagggcaatt tgtcacaaca ccttctctag aaccagcatg gataaaggcc tacaaggcgc
     7861 tctaaaaaag aagatctaaa aactataaaa aaaataatta taaaaatatc cccgtggata
     7921 agtggataac cccaagggaa gttttttcag gcatcgtgtg taagcagaat atataagtgc
     7981 tgttccctgg tgcttcctcg ctcactcgag ggcttcgccc tgtcgctcga ctgcggcgag
     8041 cactactggc tgtaaaagga cagaccacat catggttctg tgttcattag gttgttctgt
     8101 ccattgctga cataatccgc tccacttcaa cgtaacaccg cacgaagatt tctattgttc
     8161 ctgaaggcat attcaaatcg ttttcgttac cgcttgcagg catcatgaca gaacactact
     8221 tcctataaac gctacacagg ctcctgagat taataatgcg gatctctacg ataatgggag
     8281 attttcccga ctgtttcgtt cgcttctcag tggataacag ccagcttctc tgtttaacag
     8341 acaaaaacag catatccact cagttccaca tttccatata aaggccaagg catttattct
     8401 caggataatt gtttcagcat cgcaaccgca tcagactccg gcatcgcaaa ctgcacccgg
     8461 tgccgggcag ccacatccag cgcaaaaacc ttcgtgtaga cttccgttga actgatggac
     8521 ttatgtccca tcaggctttg cagaactttc agcggtatac cggcatacag catgtgcatc
     8581 gcataggaat ggcggaacgt atgtggtgtg accggaacag agaacgtcac accgtcagca
     8641 gcagcggcgg caaccgcctc cccaatccag gtcctgaccg ttctgtccgt cacttcccag
     8701 atccgcgctt ctctgtcctt cctgtgcgac ggttacgccg ctccatgagc ttatcgcgaa
     8761 taaatacctg tgacggaaga tcacttcgca gaataaataa atcctggtgt ccctgttgat
     8821 accgggaagc cctgggccaa cttttggcga aaatgagacg ttgaataact tcgtatagta
     8881 cacattatac gaagttattc ggcacgtaag aggttccaac tttcaccata atgaaataag
     8941 atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat
     9001 gcaatacgaa tggcgaaaag ccgagctcat cggtcagctt ctcaaccttg gggttacccc
     9061 cggcggtgtg ctgctggtcc acagctcctt ccgtagcgtc cggcccctcg aagatgggcc
     9121 acttggactg atcgaggccc tgcgtgctgc gctgggtccg ggagggacgc tcgtcatgcc
     9181 ctcgtggtca ggtctggacg acgagccgtt cgatcctgcc acgtcgcccg ttacaccgga
     9241 ccttggagtt gtctctgaca cattctggcg cctgccaaat gtaaagcgca gcgcccatcc
     9301 atttgccttt gcggcagcgg ggccacaggc agagcagatc atctctgatc cattgcccct
     9361 gccacctcac tcgcctgcaa gcccggtcgc ccgtgtccat gaactcgatg ggcaggtact
     9421 tctcctcggc gtgggacacg atgccaacac gacgctgcat cttgccgagt tgatggcaaa
     9481 ggttccctat ggggtgccga gacactgcac cattcttcag gatggcaagt tggtacgcgt
     9541 cgattatctc gagaatgacc actgctgtga gcgctttgcc ttggcggaca ggtggctcaa
     9601 ggagaagagc cttcagaagg aaggtccagt cggtcatgcc tttgctcggt tgatccgctc
     9661 ccgcgacatt gtggcgacag ccctgggtca actgggccga gatccgttga tcttcctgca
     9721 tccgccagag gcgggatgcg aagaatgcga tgccgctcgc cagtcgattg gctgagctca
     9781 tgagcggaga acgagatgac gttggagggg caaggtcgcg ctgattgctg gggcaacacg
     9841 tggaggcggt gtttaaacct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     9901 cggctggtac cgggcccccc ctcgaggtcg acggtatcga taagcttgat atcgaattcc
     9961 tgcagcccgg gggatccact agttctagag cggccgccac cgcggtggag ctcactcaaa
    10021 ggcggtaata cggttatcca taacttcgta taatgtatgc tatacgaagt tattagcgat
    10081 gagctcggac ttccattgtt cattccacgg acaaaaacag agaaaggaaa cgacagaggc
    10141 caaaaagctc gctttcagca cctgtcgttt cctttctttt cagctctgat gccgcatagt
    10201 taagccagta tggtcggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
    10261 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
    10321 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
    10381 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
    10441 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
    10501 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
    10561 gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
    10621 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
    10681 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
    10741 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
    10801 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
    10861 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
    10921 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
    10981 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
    11041 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
    11101 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
    11161 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
    11221 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
    11281 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
    11341 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
    11401 tcatgccata ccgcgaaagg ttttgcacca ttcgatggtg tcggatacac tccgctatcg
    11461 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
    11521 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
    11581 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctcac tcattaggca
    11641 ccccaggc
//
