LOCUS       sequence-319777-       15591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2lox landing pad plasmid with mariner transposon and transposase 
            used for CRAGE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15591)
  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 15591)
  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: pW17
FEATURES             Location/Qualifiers
     source          1..15591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1..34)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    60..167
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     84..100
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     84..100
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(134..150)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(135..151)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             376..1407
                     /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
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             1942..2736
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHTGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1996..2015)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2606..2625
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    2774..2807
                     /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)."
     primer_bind     3108..3124
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3149..3167)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     oriT            3212..3320
                     /note="incP origin of transfer"
     protein_bind    3467..3488
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3503..3508,3509..3526,3527..3533)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3541..3557
                     /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             complement(5465..6436)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(6436..7611)
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQTNHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     misc_feature    7937..8187
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(8190..8945)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      9036..9255
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      9323..9854
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      9331..9945
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     CDS             complement(13799..14248)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGI"
     primer_bind     14212..14228
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14244..14260)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14244..14260)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    14268..14284
                     /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(14292..14298,14299..14316,14317..14322))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             complement(14371..15453)
                     /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     15415..15434
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(15454..15531)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ataacttcgt atagcataca ttatacgaag ttatgataac cgtattaccg cctttgagtg
       61 agctccaccg cggtggcggc cgctctagaa ctagtggatc ccccgggctg caggaattcg
      121 atatcaagct tatcgatacc gtcgacctcg agggggggcc cggtaccgaa tgataacaaa
      181 ataaactact tttttaaaag aattttgtgt tataatctat ttattattaa gtattgggta
      241 atattttttg aagagatatt ttgaaaaaga aaaattaaag catattaaac taatttcgga
      301 ggtcattaaa actattattg aaatcatcaa actcattatg gatttaattt aaacttttta
      361 ttttaggagg caaaaatgtc caatttactg accgtacacc aaaatttgcc tgcattaccg
      421 gtcgatgcaa cgagtgatga ggttcgcaag aacctgatgg acatgttcag ggatcgccag
      481 gcgttttctg agcatacctg gaaaatgctt ctgtccgttt gccggtcgtg ggcggcatgg
      541 tgcaagttga ataaccggaa atggtttccc gcagaacctg aagatgttcg cgattatctt
      601 ctatatcttc aggcgcgcgg tctggcagta aaaactatcc agcaacattt gggccagcta
      661 aacatgcttc atcgtcggtc cgggctgcca cgaccaagtg acagcaatgc tgtttcactg
      721 gttatgcggc ggatccgaaa agaaaacgtt gatgccggtg aacgtgcaaa acaggctcta
      781 gcgttcgaac gcactgattt cgaccaggtt cgttcactca tggaaaatag cgatcgctgc
      841 caggatatac gtaatctggc atttctgggg attgcttata acaccctgtt acgtatagcc
      901 gaaattgcca ggatcagggt taaagatatc tcacgtactg acggtgggag aatgttaatc
      961 catattggca gaacgaaaac gctggttagc accgcaggtg tagagaaggc acttagcctg
     1021 ggggtaacta aactggtcga gcgatggatt tccgtctctg gtgtagctga tgatccgaat
     1081 aactacctgt tttgccgggt cagaaaaaat ggtgttgccg cgccatctgc caccagccag
     1141 ctatcaactc gcgccctgga agggattttt gaagcaactc atcgattgat ttacggcgct
     1201 aaggatgact ctggtcagag atacctggcc tggtctggac acagtgcccg tgtcggagcc
     1261 gcgcgagata tggcccgcgc tggagtttca ataccggaga tcatgcaagc tggtggctgg
     1321 accaatgtaa atattgtcat gaactatatc cgtaacctgg atagtgaaac aggggcaatg
     1381 gtgcgcctgc tggaagatgg cgattagccc agcccgccta atgagcgggc ttttttttga
     1441 acaaaactcg agacagtaag acgggtaagc ctgttgatga taccgctgcc ttactgggtg
     1501 cattagccag tctgaatgac ctgtcacggg ataatccgaa gtggtcagac tggaaaatca
     1561 gagggcagga actgctgaac cttaactagt tcacgctgcc gcaagcactc agggcgcaag
     1621 ggctgctaaa ggaagcggaa cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg
     1681 atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa gagaaagcag
     1741 gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg gacagcaagc
     1801 gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac
     1861 tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagatc tgatcaagag
     1921 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacacagg ttctccggcc
     1981 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     2041 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     2101 tccggtgccc tgaatgaact ccaagacgag gcagcgcggc tatcgtggct ggccacgacg
     2161 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     2221 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     2281 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     2341 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     2401 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     2461 ctcaaggcgc ggatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     2521 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     2581 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     2641 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     2701 atcgccttct atcgccttct tgacgagttc ttctgagcgg cccgggtgaa agttggaacc
     2761 tcttacgtgc cgaataactt cgtataatgt gtactatacg aagttattca acgtctcatt
     2821 ttcgccaaaa gttggcccag gacttcccgg tatcaacagt actgcgatga gtggcagggc
     2881 ggggcgtaat ttttttaagg cagttattgg tgcctagaaa tattttatct gattaataag
     2941 atgatcttct tgagatcgtt ttggtctgcg cgtgtcgacg actctggggt tcgcgacact
     3001 ggcagagcat tacgccctgc aggatgcaat gggccggcca gaccggggac ttatcagcca
     3061 acctgttatg tgttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     3121 cagtgaatta attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     3181 gtcatgataa taatggtttc ttagagctta tcggccagcc tcgcagagca ggattcccgt
     3241 tgagcaccgc caggtgcgaa taagggacag tgaagaagga acacccgctc gcgggtgggc
     3301 ctacttcacc tatcctgccc ggctgacgcc gttggataca ccaaggaaag tctacacgaa
     3361 ccctttggca aaatcctgta tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta
     3421 taatgacccc gaagcagggt tatgcagcgg gtgagcgcaa cgcaattaat gtgagttagc
     3481 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     3541 ttgtgagcgg ataacaattt cacacaggaa acacatatgg aaaaaaagga atttcgtgtt
     3601 ttgataaaat actgttttct gaagggaaaa aatacagtgg aagcaaaaac ttggcttgat
     3661 aatgagtttc cggactctgc cccagggaaa tcaacaataa ttgattggta tgcaaaattc
     3721 aagcgtggtg aaatgagcac ggaggacggt gaacgcagtg gacgcccgaa agaggtggtt
     3781 accgacgaaa acatcaaaaa aatccacaaa atgattttga atgaccgtaa aatgaagttg
     3841 atcgagatag cagaggcctt aaagatatca aaggaacgtg ttggtcatat cattcatcaa
     3901 tatttggata tgcggaagct ctgtgcaaaa tgggtgccgc gcgagctcac atttgaccaa
     3961 aaacaacaac gtgttgatga ttctgagcgg tgtttgcagc tgttaactcg taatacaccc
     4021 gagtttttcc gtcgatatgt gacaatggat gaaacatggc tccatcacta cactcctgag
     4081 tccaatcgac agtcggctga gtggacagcg accggtgaac cgtctccgaa gcgtggaaag
     4141 actcaaaagt ccgctggcaa agtaatggcc tctgtttttt gggatgcgca tggaataatt
     4201 tttatcgatt atcttgagaa gggaaaaacc atcaacagtg actattatat ggcgttattg
     4261 gagcgtttga aggtcgaaat cgcggcaaaa cggccccaca tgaagaagaa aaaagtgttg
     4321 ttccaccaag acaacgcacc gtgccacaag tcattgagaa cgatggcaaa aattcatgaa
     4381 ttgggcttcg aattgcttcc ccacccgccg tattctccag atctggcccc cagcgacttt
     4441 ttcttgttct cagacctcaa aaggatgctc gcagggaaaa aatttggctg caatgaagag
     4501 gtgatcgccg aaactgaggc ctattttgag gcaaaaccga aggagtacta ccaaaatggt
     4561 atcaaaaaat tggaaggtcg ttataatcgt tgtatcgctc ttgaagggaa ctatgttgaa
     4621 taataaaaac gaattttcac aaaaaaatgt gtttttcttt gttagaccgg atctgcagaa
     4681 ttccagcaca ctggcggccg ttactagatg catgctcgag cggccgccag tgtgatggat
     4741 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4801 ctcgtcgaca gcgacacact tgcatcggat gcagcccggt taacgtgccg gcacggcctg
     4861 ggtaaccagg tattttgtcc acataaccgt gcgcaaaatg ttgtggataa gcaggacaca
     4921 gcagcaatcc acagcaggca tacaaccgca caccgaggtt actccgttct acaggttacg
     4981 acgacatgtc aatacttgcc cttgataggc attgatggaa tcgtagtctc acgctgatag
     5041 tctgatcgac aatacaagtg ggaccgtggt cccagaccga taatcagacc gacaacacga
     5101 gtgggatcgt ggtcccagac taataatcag accgacgata cgagtgggac cgtggtccca
     5161 gactaataat cagaccgacg atacgagtgg gaccgtggty ccagwctrat watcagaccg
     5221 acgatacgag tgggaccgtg gtcccagact aataatcaga ccgacgatac gagtgggacc
     5281 atggtcccag actaataatc agaccgacga tacgagtggg accgtggtcc cagtctgatt
     5341 atcagaccga cgatacaagt ggaacagtgg gcccagagag aatattcagg ccagttatgc
     5401 tttctggcct gtaacaaagg acattaagta aagacagata aacgtagact aaaacgtggt
     5461 cgcatcaggg tgctggcttt tcaagttcct taagaatggc ctcaattttc tctatacact
     5521 cagttggaac acgagacctg tccaggttaa gcaccatttt atcgccctta tacaatactg
     5581 tcgctccagg agcaaactga tgtcgtgagc ttaaactagt tcttgatgca gatgacgttt
     5641 taagcacaga agttaaaaga gtgataactt cttcagcttc aaatatcacc ccagcttttt
     5701 tctgctcatg aaggttagat gcctgctgct taagtaattc ctctttatct gtaaaggctt
     5761 tttgaagtgc atcacctgac cgggcagata gttcaccggg gtgagaaaaa agagcaacaa
     5821 ctgatttagg caatttggcg gtgttgatac agcgggtaat aatcttacgt gaaatatttt
     5881 ccgcatcagc cagcgcagaa atatttccag caaattcatt ctgcaatcgg cttgcataac
     5941 gctgaccacg ttcataagca cttgttgggc gataatcgtt acccaatctg gataatgcag
     6001 ccatctgctc atcatccagc tcgccaacca gaacacgata atcactttcg gtaagtgcag
     6061 cagctttacg acggcgactc ccatcggcaa tttctatgac accagatact cttcgaccga
     6121 acgccggtgt ctgttgacca gtcagtagaa aagaagggat gagatcatcc agtgcgtcct
     6181 cagtaagcag ctcctggtca cgttcattac ctgaccatac ccgagaggtc ttctcaacac
     6241 tatcaccccg gagcacttca agagtaaact tcacatcccg accacataca ggcaaagtaa
     6301 tggcattacc gcgagccatt actcctacgc gcgcaattaa cgaatccacc atcggggcag
     6361 ctggtgtcga taacgaagta tcttcaaccg gttgagtatt gagcgtatgt tttggaataa
     6421 caggcgcacg cttcattatc taatctccca gcgtggttta atcagacgat cgaaaatttc
     6481 attgcagaca ggttcccaaa tagaaagagc atttctccag gcaccagttg aagagcgttg
     6541 atcaatggcc tgttcaaaaa cagttctcat ccggatctga cctttaccaa cttcatccgt
     6601 ttcacgtaca acatttttta gaaccatgct tccccaggca tcccgaattt gctcctccat
     6661 ccacggggac tgagagccat tactattgct gtatttggta agcaaaatac gtacatcagg
     6721 ctcgaaccct ttaagatcaa cgttcttgag cagatcacga agcatatcga aaaactgcag
     6781 tgcggaggtg tagtcaaaca actcagcagg cgtgggaaca atcagcacat cagcagcaca
     6841 tacgacatta atcgtgccga tacccaggtt aggcgcgctg tcaataacta tgacatcata
     6901 gtcatgagca acagtttcaa tggccagtcg gagcatcagg tgtggatcgg tgggcagttt
     6961 accttcatca aatttgccca ttaactcagt ttcaatacgg tgcagagcca gacaggaagg
     7021 aataatgtca agccccggcc agcaagtggg ctttattgca taagtgacat cgtccttttc
     7081 cccaagatag aaaggcagga gagtgtcttc tgcatgaata tgaagatctg gtacccatcc
     7141 gtgatacatt gaggctgttc cctgggggtc gttaccttcc acgagcaaaa cacgtagccc
     7201 cttcagagcc agatcctgag caagatgaac agaaactgag gttttataaa cgccaccttt
     7261 atgggcagca accccgatca ccggtggaaa tacgtcttca gcacgtcgca atcgcgtacc
     7321 aaacacatca cgcatatgat tagtttgttc aattgtataa ccaacacgtt gctcaacccg
     7381 tcctcgaatt tccatatccg ggtgcggtag tcgccctgct ttctcggcat ctctgatagc
     7441 ctgagaagaa accccaacta aatccgctgc ttcacctatt ctccagcgcc gggttatttt
     7501 cctcgcttcc gggctgtcat cattaaactg tgcaatggcg atagccttcg tcatttcatg
     7561 accagcgttt atgcactggt taagtgtttc catgagtttc attctgaaca tcctttaatc
     7621 attgctttgc gtttttttat taaatcttgc aatttactgc aaagcaacaa caaaatcgca
     7681 aagtcatcaa aaaaccgcaa agttgtttaa aataagagca acactacaaa aggagataag
     7741 aagagcacat acctcagtca cttattatca ctagcgctcg ccgcagccgt gtaaccgagc
     7801 atagcgagcg aactggcgag gaagcaaaga agaactgttc tgtcagatag ctcttacgct
     7861 cagcgcaaga agaaatatcc accgtgggaa aaactccagg tagaggtaca cacgcggata
     7921 gccaattcag agtaataaac tgtgataatc aaccctcatc aatgatgacg aactaacccc
     7981 cgatatcaag tcacatgacg aagggaaaga gaaggaaatc aactgtgaca aactgccctc
     8041 aaatttggct tccttaaaaa ttacagttca aaaagtatga gaaaatccat gcaggctgaa
     8101 ggaaacagca aaactgtgac aaattaccct cagtaggtca gaacaaatgt gacgaaccac
     8161 cctcaaatct gtgacagata accctcagac tatcctgtcg tcatggaagt gatatcgcgg
     8221 aaggaaaata cgatatgagt cgtctggcgg cctttctttt tctcaatgta tgagaggcgc
     8281 attggagttc tgctgttgat ctcattaaca cagacctgca ggaagcggcg gcggaagtca
     8341 ggcatacgct ggtaactttg aggcagctgg taacgctcta tgatccagtc gattttcaga
     8401 gagacgatgc ctgagccatc cggcttacga tactgacaca gggattcgta taaacgcatg
     8461 gcatacggat tggtgatttc ttttgtttca ctaagccgaa actgcgtaaa ccggttctgt
     8521 aacccgataa agaagggaat gagatatggg ttgatatgta cactgtaaag ccctctggat
     8581 ggactgtgcg cacgtttgat aaaccaagga aaagattcat agcctttttc atcgccggca
     8641 tcctcttcag ggcgataaaa aaccacttcc ttccccgcga aactcttcaa tgcctgccgt
     8701 atatccttac tggcttccgc agaggtcaat ccgaatattt cagcatattt agcaacatgg
     8761 atctcgcaga taccgtcatg ttcctgtagg gtgccatcag attttctgat ctggtcaacg
     8821 aacagataca gcatacgttt ttgatcccgg gagagactat atgccgcctc agtgaggtcg
     8881 tttgactgga cgattcgcgg gctattttta cgtttcttgt gattgataac cgctgtttcc
     8941 gccatgacag atccatgtga agtgtgacaa gtttttagat tgtcacacta aataaaaaag
     9001 agtcaataag cagggataac tttgtgaaaa aacagcttct tctgagggca atttgtcaca
     9061 gggttaaggg caatttgtca cagacaggac tgtcatttga gggtgatttg tcacactgaa
     9121 agggcaattt gtcacaacac cttctctaga accagcatgg ataaaggcct acaaggcgct
     9181 ctaaaaaaga agatctaaaa actataaaaa aaataattat aaaaatatcc ccgtggataa
     9241 gtggataacc ccaagggaag ttttttcagg catcgtgtgt aagcagaata tataagtgct
     9301 gttccctggt gcttcctcgc tcactcgacc gggagggttc gagaaggggg ggcacccccc
     9361 ttcggcgtgc gcggtcacgc gcacagggcg cagccctggt taaaaacaag gtttataaat
     9421 attggtttaa aagcaggtta aaagacaggt tagcggtggc cgaaaaacgg gcggaaaccc
     9481 ttgcaaatgc tggattttct gcctgtggac agcccctcaa atgtcaatag gtgcgcccct
     9541 catctgtcag cactctgccc ctcaagtgtc aaggatcgcg cccctcatct gtcagtagtc
     9601 gcgcccctca agtgtcaata ccgcagggca cttatcccca ggcttgtcca catcatctgt
     9661 gggaaactcg cgtaaaatca ggcgttttcg ccgatttgcg aggctggcca gctccacgtc
     9721 gccggccgaa atcgagcctg cccctcatct gtcaacgccg cgccgggtga gtcggcccct
     9781 caagtgtcaa cgtccgcccc tcatctgtca gtgagggcca agttttccgc gaggtatcca
     9841 caacgccggc ggccggccgc ggtgtctcgc acacggcttc gacggcgttt ctggcgcgtt
     9901 tgcagggcca tagacggccg ccagcccagc ggcgagggca accagccgag ggcttcgccc
     9961 tgtcgctcga ctgcggcgag cactactggc tgtaaaagga cagaccacat catggttctg
    10021 tgttcattag gttgttctgt ccattgctga cataatccgc tccacttcaa cgtaacaccg
    10081 cacgaagatt tctattgttc ctgaaggcat attcaaatcg ttttcgttac cgcttgcagg
    10141 catcatgaca gaacactact tcctataaac gctacacagg ctcctgagat taataatgcg
    10201 gatctctacg ataatgggag attttcccga ctgtttcgtt cgcttctcag tggataacag
    10261 ccagcttctc tgtttaacag acaaaaacag catatccact cagttccaca tttccatata
    10321 aaggccaagg catttattct caggataatt gtttcagcat cgcaaccgca tcagactccg
    10381 gcatcgcaaa ctgcacccgg tgccgggcag ccacatccag cgcaaaaacc ttcgtgtaga
    10441 cttccgttga actgatggac ttatgtccca tcaggctttg cagaactttc agcggtatac
    10501 cggcatacag catgtgcatc gcataggaat ggcggaacgt atgtggtgtg accggaacag
    10561 agaacgtcac accgtcagca gcagcggcgg caaccgcctc cccaatccag gtcctgaccg
    10621 ttctgtccgt cacttcccag atccgcgctt tctctgtcct tcctgtgcga cggttacgcc
    10681 gctccatgag cttatcgcga ataaatacct gtgacggaag atcacttcgc agaataaata
    10741 aatcctggtg tccctgtgca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
    10801 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
    10861 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
    10921 ttaaagttct gctaacaggt tggctgataa gtccccggtc tgacacatag atggcgtcgc
    10981 tagtattaaa tgcatattat ttttatatag taccaacctt caaatgattc ccagatctca
    11041 aactggaaca acactcaacc ctatctcggt cgcgaccaca cccgtcctgt ggatccggag
    11101 tcgtattgat ttggcgttac gcgaacgcga agtccgactc taagatgtca cggaggttca
    11161 agttaccttt agccggaagt gctggcattt tgtccaattg agactcgtgc aactggtcag
    11221 cgaactggtc gtagaaatca gccagtacat cacaagactc atatgtgtca accatagttt
    11281 cgcgcactgc tttgaacagg ttcgcagcgt cagccggaat ggtaccgaag gagtcgtgaa
    11341 tcagtgcaaa agattcgatt ccgtacttct cgtgtgccca cactacagtc ttacgaaggt
    11401 ggctaccgtc ttggctgtgt acaaagttag gagcgatacc agactcctgt ttgtgtgcat
    11461 caatctcgct atctttgttg gtgttaatgg taggctgtaa gcggaactga ccgaggaaca
    11521 tcaggttcaa gcgcgtctga ataggcttct tgtattcctg ccacacaggg aaaccatcag
    11581 gagttaccca atgcacagcg caacgcttgc gaagaatctc tccagtcttc ttatctttga
    11641 cctcagcagc cagcagctta gcagcagact taagccagtt cattgcttca accgcagcta
    11701 ccaccgtcac gctcacagat tcccaaatca gcttagccat gtatccagca gcctgattcg
    11761 gctgagtgaa catcagaccc ttgccggaat caatagctgg ctgaatggta tcttccagca
    11821 cttgttgacg gaagccgaac tctttggacc cgtaagccag cgtcatgact gaacgcttag
    11881 tcacactgcg agtaacaccg taagccagcc attgaccagc cagtgcctta gtgcccagct
    11941 tgactttctc agagatttca ccagtgttct catcggtcac ggtaactact tcgttatcgg
    12001 tcccattgat tgcgtctgct tgtagaatct cgttgacttt cttagcaaca atcccgtaga
    12061 tgtcctgaac ggtttcacta ggaagcaagt taaccgcgcg accacctacc tcatctcgga
    12121 gcatcgcgga gaagtgctgg atgccagagc aagacccgtc aaacgccagc ggaagggagc
    12181 agttatagct caggccgtgg tgctgtaccc cagcgtactc aaagcagaac gcaaggaagc
    12241 agaacggaga atcttgctca gcccaccaag tgttctccag tggagactta gcgcaagcca
    12301 tgatgttctc gtggttttcc tcaatgaact tgatgcgctc tgggaacgga accttatcga
    12361 cacccgcaca gtttgcaccg tggattttca gccagtagta accttcctta ccgattggtt
    12421 tacctttcgc cagcgtaagc agtcctttgg tcatatcgtt accttgcggg ttgaacattg
    12481 acacagcgta aacacgaccg cgccagtcca tgttgtaagg gaaccagatg gccttatggt
    12541 tagcaaactt attggcttgc tcaagcatga actcaaggct gatacggcga gacttgcgag
    12601 ccttgtcctt gcggtacaca gcagcggcag cacgtttcca cgcggtgaga gcctcaggat
    12661 tcatgtcgat gtcttccggt ttcatcggga gttcttcacg ctcaatcgca gggatgtcct
    12721 cgaccggaca atgcttccac ttggtgatta cgttggcgac cgctaggact ttcttgttga
    12781 ttttccatgc ggtgttttgc gcaatgttaa tcgctttgta cacctcaggc atgtaaacgt
    12841 cttcgtagcg catcagtgct ttcttactgt gagtacgcac cagcgccaga ggacgacgac
    12901 cgttagccca atagccacca ccagtaatgc cagtccacgg cttaggagga actacgcaag
    12961 gttggaacat cggagagatg ccagccagcg cacctgcacg ggttgcgata gcctcagcgt
    13021 attcaggtgc gagttcgata gtctcagagt cttgacctac tacgccagca ttttggcggt
    13081 gtaagctaac cattccggtt gactcaatga gcatctcgat gcagcgtact cctacatgaa
    13141 tagagtcttc cttatgccac gaagaccacg cctcgccacc gagtagaccc ttagagagca
    13201 tgtcagcctc gacaacttgc ataaatgctt tcttgtagac gtgccctacg cgcttgttga
    13261 gttgttcctc aacgtttttc ttgaagtgct tagcttcaag gtcacggata cgaccgaagc
    13321 gagcctcgtc ctcaatggcc cgaccgattg cgcttgctac agcctgaacg gttgtattgt
    13381 cagcactggt taggcaagcc agagtggtct taatggtgat gtacgctacg gcttccggct
    13441 tgatttcttg caggaactgg aaggctgtcg ggcgcttgcc gcgcttagct ttcacttcct
    13501 caaaccagtc gttgatgcgt gcaatcatct tagggagtag ggtagtgatg agaggcttgg
    13561 cggcagcgtt atccgcaacc tcaccagctt taagttgacg ctcaaacatc ttgcggaagc
    13621 gtgcttcacc catctcgtaa gactcatgct caagggccaa ctgttcgcga gctaaacgct
    13681 caccgtaatg gtcagccaga gtgttgaacg ggatagcagc cagttcgatg tcagagaagt
    13741 cgttcttagc gatgttaatc gtgttcattt agtgcctctt ccagttagta aatccggatc
    13801 agatcccgac gccatcaaaa ataattcgcg tctggccttc ctgtagccag ctttcatcaa
    13861 cattaaatgt gagcgagtaa caacccgtcg gattctccgt gggaacaaac ggcggattga
    13921 ccgtaatggg ataggtcacg ttggtgtaga tgggcgcatc gtaaccgtgc atctgccagt
    13981 ttgaggggac gacgacagta tcggcctcag gaagatcgca ctccagccag ctttccggca
    14041 ccgcttctgg tgccggaaac caggcaaagc gccattcgcc attcaggctg cgcaactgtt
    14101 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
    14161 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
    14221 cggccagtga atccgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
    14281 acaattccac acattatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
    14341 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
    14401 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
    14461 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
    14521 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
    14581 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
    14641 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
    14701 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
    14761 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
    14821 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
    14881 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
    14941 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
    15001 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
    15061 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
    15121 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
    15181 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
    15241 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
    15301 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
    15361 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
    15421 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
    15481 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctg
    15541 gatcctctac gccggacgca tcgtgcaaag cctcgcaatc cagtgcaaag g
//
