LOCUS       sequence-474333-       16144 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transmissible episome that contains the ShBle cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16144)
  TITLE     Spheroplasted cells: a game changer for DNA delivery to diatoms
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 16144)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pPtGE30
FEATURES             Location/Qualifiers
     source          1..16144
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             604..978
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        1675..1890
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             1891..2694
                     /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
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     oriT            3453..3562
                     /note="incP origin of transfer"
     CDS             3595..3966
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      6637..7251
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      6728..7259
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      7327..7546
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             7637..8392
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    8395..8645
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     misc_feature    8807..9310
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        9311..9497
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             9498..10160
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     CDS             10350..11525
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             11525..12496
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    12569..13042
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     misc_feature    13302..13700
                     /label=cos
                     /note="lambda cos site; allows packaging into phage lambda 
                     particles"
     protein_bind    complement(13718..13751)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     13819..13838
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(13918..14193)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGETIEYSSLHACRSTLEDPRVPSSNSPYSESYYNSL
                     AVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS"
     primer_bind     14031..14053
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     14045..14062
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     14046..14062
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     14069..14088
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        14069..14087
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    14090..14146
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(14189..14205)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14189..14205)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14202..14224)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14213..14229
                     /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(14237..14243,14244..14261,14262..14267))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14282..14303
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(14523..15182)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     15097..15116
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(15183..15285)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 actagcttga ttgggatatc tcgctcgtgc ttgtcgcgtg ctatgtcttt ttagggtact
       61 ttgaacctac gttcgtactt gtgtaatatg atcatcgtcg tcatcgtatt atcgtttttc
      121 atccgtccag cgcaaaatgc attagcagct agtcctagcg tgcggagcta cctgtacagg
      181 tgcatgacgg atgcgtgtcc ttaagtgagt ttctaattaa cagtaacttc tttacttatg
      241 tttcagtttg taagaagcgg gatgcgctcg tcgcttgaca tctgattgga ctgcgtcggc
      301 acgtgaaaac tacattgtga aatctgctaa aactccgggt atctctgaca caaaacgatt
      361 cggcttcgca atttcaacat tacggtcaag gctaacgtat ctttctcggt caacttcaga
      421 ttatgccgat taaattgtcg tagctttcaa ggcgttttga gtactgcggc agttgttgaa
      481 cctgcaagga gaagatctcg acaacagaat aaagcgaaaa atgggtctca tgcactaaca
      541 ctcaggcctc cctcataatc tctgtttgag tttaccaaca acacatatat acatttcgac
      601 aaaatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg
      661 gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc
      721 ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgcca
      781 gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg
      841 gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag
      901 cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg
      961 gccgaggagc aggactgacc gacgccgacc aacaccgccg gtccgacgcg gcccgacggg
     1021 tccgaggcct cggagatctg ggcccatgcg gccgcaacaa ctacctcgac tttggctggg
     1081 acactttcag tgaggacaag aagcttcaga agcgtgctat cgaactcaac cagggacgtg
     1141 cggcacaaat gggcatcctt gctctcatgg tgcacgaaca gttgggagtc tctatccttc
     1201 cttaaaaatt taattttcat tagttgcagt cactccgctt tggtttcgta actataacgg
     1261 tcctaaggta gcgaacgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg
     1321 gacagcttca aggatcgctc gcggctctta ccagcctaac ttcgatcatt ggaccgctga
     1381 tcgtcacggc gatttatgcc gcctcggcga gcacatggaa cgggttggca tggattgtag
     1441 gcgccgccct ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca
     1501 cctcgacctg aatggaagcc ggcggcacct cgctaacgga ttcaccactc caagaattgg
     1561 agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc aacccttggc agaacatatc
     1621 catcgcgtcc gccatctcca gcagccgcac gcggcgcatc gggggggggg gggtttcaat
     1681 tcatcatttt ttttttattc ttttttttga tttcggtttc cttgaaattt ttttgattcg
     1741 gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata
     1801 cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga
     1861 acaaaaacct gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct
     1921 gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca
     1981 aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca
     2041 ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg
     2101 gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgaa
     2161 gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac
     2221 agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt
     2281 agcggtttga agcaggcggc ggaagaagta acaaaggaac ctagaggcct tttgatgtta
     2341 gcagaattgt catgcaaggg ctccctagct actggagaat atactaaggg tactgttgac
     2401 attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga
     2461 agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga
     2521 gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg tctctacagg atctgacatt
     2581 attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt
     2641 tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact
     2701 gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat
     2761 cagttattac ccggccggga atctcggtcg taatgatttt tataatgacg aaaaaaaaaa
     2821 aaattggaaa gaaaaccccc cccccccccg cagcgttggg tcctggccac gggtgcgcat
     2881 gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca
     2941 gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac
     3001 ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa
     3061 gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg
     3121 tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg
     3181 gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg
     3241 ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcgagt tacgctaggg
     3301 ataacagggt aatatagatc ttccgctgca taaccctgct tcggggtcat tatagcgatt
     3361 ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag ggttcgtgta
     3421 gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt aggcccaccc
     3481 gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct caacgggaat
     3541 cctgctctgc gaggctggcc ggctaccgcc ggcgtaacag atgagggcaa gcggatggct
     3601 gatgaaacca agccaaccag gaagggcagc ccacctatca aggtgtactg ccttccagac
     3661 gaacgaagag cgattgagga aaaggcggcg gcggccggca tgagcctgtc ggcctacctg
     3721 ctggccgtcg gccagggcta caaaatcacg ggcgtcgtgg actatgagca cgtccgcgag
     3781 ctggcccgca tcaatggcga cctgggccgc ctgggcggcc tgctgaaact ctggctcacc
     3841 gacgacccgc gcacggcgcg gttcggtgat gccacgatcc tcgccctgct ggcgaagatc
     3901 gaagagaagc aggacgagct tggcaaggtc atgatgggcg tggtccgccc gagggcagag
     3961 ccatgacttt tttagccgct aaaacggccg gggggtgcgc gtgattgcca agcacgtccc
     4021 catgcgctcc atcaagaaga ggcacttcga gctgtaagta catcaccgac gagcaaggca
     4081 agacgatccg cgcctgtttt attgagaacg ttgttcgtgt tggcctcaat ggtagcgatg
     4141 cgtcattcag cgaagttctg gtgctgatga tgtggttcgc tttgccactg gtcaatgtgg
     4201 taagcccgtg taatgtcagt aaccttttta ctgatctcag cttgagcacg gtcgctgatg
     4261 agcttatcca tggccccacg gtaacggata tgatcctcta gggcgttgac aaattctttg
     4321 tcggttttca tggggtagac gtcagtgacc agggaacgtc tccctacaaa aagttgcgca
     4381 tactttgctc cgttgtcgac ggcaggggtg tcggaccaga ttgtatctgt ggcaacggcc
     4441 tcattgcgtc aatggacctt taaagccggg aaacgagact tgaaatgttt acgtagaggt
     4501 gcattataga cctcacgcgc atattgagtg gttgcaaaaa tggttttgcg aacggtatca
     4561 ctggagaccc atgccaggca aggacggagg gcatcataat catgttcatt gcgttggaca
     4621 gcatgcttgt tacacgtaag tatacggtcg agagcatcgt gacgttgaga attacagatg
     4681 gcaacgtgtc ggtgcatgta ttggccgaga ttgtcaaatc gtggctcaat atatggcatg
     4741 ccaggtaagt catggatgtc ggtatgccat tcagccttca tgtcgatttc gttgtcgaga
     4801 actgatgggt cccagtcaac gtcagaagta agaacaacat ggggaaggga atgaagttca
     4861 tgactggtgg gtgcgcgcat atccatgtac ggaagaccct gtcgtatgtt tagtgggatg
     4921 atgtaatcat ccagggtaac aatacgttgg ttgccgccca cggtacgaga ccgatcttga
     4981 acacaattgt ggcaatgttc aagttgggcg ctggaatgga tggttttacc tttaccaaga
     5041 tgcgcgtatt gatgcatgat aacaacaatg ggccctcgtt gcgattcgac aagaccggca
     5101 gccgtgacaa tgtctagatt ggacaaggta tgttcattta taccggttat gttggctgaa
     5161 cgtccagttt tgtgcagtac ggtaacatca ctccccgcaa gtccaccgtt ggcaccgcga
     5221 tcgacaagtg cagatgtggt attttgtact gtgtgtcggg agacttggta ctgtaacaca
     5281 tttgactgaa gtgacttggt ggtaccatga ggacggaggg aacccgttga tacggtgttg
     5341 cgtgacgctg caagtacctt acgtatgtcg cctggatcca tacgtccaac ccgatcagaa
     5401 agatgtgcta ggagttccgt ttctggtgca caatcatgaa aggtatccac gggagttcca
     5461 ttgttgtcgt ccggggctgc gtcgcgactg ccagttgcat gtacatgcgc catcaaggga
     5521 cgtgccaatc cattactaac cgggagaacc ttagctgggg cagccaatcc ctgaagtata
     5581 gccttagcat catctgagag ctggttccac atatctctag ggatataagg tcgttcacgg
     5641 ttagtgggcg tactatgatg tcggttatta gtacaattct ccctgcgggc atgcgcattg
     5701 gcttcataga gtacggaagg tgacaagtca atattgtagt taacatccgg atcagtgtca
     5761 aagtctgtag gatggtaaga aagatcagta gaatgaatac tacgcttttc cttgggacta
     5821 cgggaattag agaagttgtt tcctttattg tagagtgacg ctgaagcaag tagaagacta
     5881 aggtaactct cgtagctaat aggattacct cctttggcta ggtcaagagt ggctgtgatc
     5941 ttcacttgac aaagttccgg tacattgtgg acagcattct ccaaaagact aagacacagt
     6001 tgctttggga gttgctcagc cattggtaca gtattgtggt agatacaaag gtggttcttc
     6061 caatgaagga taaatccttc tgctgtgcct gtccatgagg atccatattt cgccgtagtt
     6121 aggtaaccaa gcgtggtggc tgaactgatc ttcgcacttg ctgattccgt atagtgtttg
     6181 acaactttac aaaacacttc ttgcgcagtt cgctctagcc tggacagaga agggacaatc
     6241 ttcgtcgtta agactcgtgc gaatagcaaa agatcacaaa atagcacatc ggcaccgacc
     6301 aacgattatt tccaaggaaa aaaagaatgc ttcactacaa gaaattgtgt catccctata
     6361 cagagtcttg ttactgtgac agaaaattga tggaagatgt ggcggattgc ctttacacta
     6421 gccaacttgt tcgactaatt gcagcttctt ctgagaggct tcaccgagta acgcgaagaa
     6481 caccggtgtc tcgtacatgc tcgtcggtga acgctcgtcc aatgacaccc cccactttgt
     6541 atcaatatcc caacttggta gtgaactgga atgatacatg caatttcgcg ccgcatcaac
     6601 agccacgggc accatcgacg aatagactcg gtcgagctgg ttgccctcgc cgctgggctg
     6661 gcggccgtct atggccctgc aaacgcgcca gaaacgccgt cgaagccgtg tgcgagacac
     6721 cgcggccggc cgccggcgtt gtggatacct cgcggaaaac ttggccctca ctgacagatg
     6781 aggggcggac gttgacactt gaggggccga ctcacccggc gcggcgttga cagatgaggg
     6841 gcaggctcga tttcggccgg cgacgtggag ctggccagcc tcgcaaatcg gcgaaaacgc
     6901 ctgattttac gcgagtttcc cacagatgat gtggacaagc ctggggataa gtgccctgcg
     6961 gtattgacac ttgaggggcg cgactactga cagatgaggg gcgcgatcct tgacacttga
     7021 ggggcagagt gctgacagat gaggggcgca cctattgaca tttgaggggc tgtccacagg
     7081 cagaaaatcc agcatttgca agggtttccg cccgtttttc ggccaccgct aacctgtctt
     7141 ttaacctgct tttaaaccaa tatttataaa ccttgttttt aaccagggct gcgccctgtg
     7201 cgcgtgaccg cgcacgccga aggggggtgc ccccccttct cgaaccctcc cggtcgagtg
     7261 agcgaggaag caccagggaa cagcacttat atattctgct tacacacgat gcctgaaaaa
     7321 acttcccttg gggttatcca cttatccacg gggatatttt tataattatt ttttttatag
     7381 tttttagatc ttctttttta gagcgccttg taggccttta tccatgctgg ttctagagaa
     7441 ggtgttgtga caaattgccc tttcagtgtg acaaatcacc ctcaaatgac agtcctgtct
     7501 gtgacaaatt gcccttaacc ctgtgacaaa ttgccctcag aagaagctgt tttttcacaa
     7561 agttatccct gcttattgac tcttttttat ttagtgtgac aatctaaaaa cttgtcacac
     7621 ttcacatgga tctgtcatgg cggaaacagc ggttatcaat cacaagaaac gtaaaaatag
     7681 cccgcgaatc gtccagtcaa acgacctcac tgaggcggca tatagtctct cccgggatca
     7741 aaaacgtatg ctgtatctgt tcgttgacca gatcagaaaa tctgatggca ccctacagga
     7801 acatgacggt atctgcgaga tccatgttgc taaatatgct gaaatattcg gattgacctc
     7861 tgcggaagcc agtaaggata tacggcaggc attgaagagt ttcgcgggga aggaagtggt
     7921 tttttatcgc cctgaagagg atgccggcga tgaaaaaggc tatgaatctt ttccttggtt
     7981 tatcaaacgt gcgcacagtc catccagagg gctttacagt gtacatatca acccatatct
     8041 cattcccttc tttatcgggt tacagaaccg gtttacgcag tttcggctta gtgaaacaaa
     8101 agaaatcacc aatccgtatg ccatgcgttt atacgaatcc ctgtgtcagt atcgtaagcc
     8161 ggatggctca ggcatcgtct ctctgaaaat cgactggatc atagagcgtt accagctgcc
     8221 tcaaagttac cagcgtatgc ctgacttccg ccgccgcttc ctgcaggtct gtgttaatga
     8281 gatcaacagc agaactccaa tgcgcctctc atacattgag aaaaagaaag gccgccagac
     8341 gactcatatc gtattttcct tccgcgatat cacttccatg acgacaggat agtctgaggg
     8401 ttatctgtca cagatttgag ggtggttcgt cacatttgtt ctgacctact gagggtaatt
     8461 tgtcacagtt ttgctgtttc cttcagcctg catggatttt ctcatacttt ttgaactgta
     8521 atttttaagg aagccaaatt tgagggcagt ttgtcacagt tgatttcctt ctctttccct
     8581 tcgtcatgtg acctgatatc gggggttagt tcgtcatcat tgatgagggt tgattatcac
     8641 agtttattac tctgaattgg ctatccgcgt gtgtacctct acctggagtt tttcccacgg
     8701 tggatatttc ttcttgcgct gagcgtaaga gctatctgac agaacagttc ttctttgctt
     8761 cctcgccagt tcgctcgcta tgctcggtta cacggctgcg gcgagcatca cgtgctataa
     8821 aaataattat aatttaaatt ttttaatata aatatataaa ttaaaaatag aaagtaaaaa
     8881 aagaaattaa agaaaaaata gtttttgttt tccgaagatg taaaagactc tagggggatc
     8941 gccaacaaat actacctttt accttgctct tcctgctctc aggtattaat gccgaattgt
     9001 ttcatcttgt ctgtgtagaa gaccacacac gaaaatcctg tgattttaca ttttacttat
     9061 cgttaatcga atgtatatct atttaatctg cttttcttgt ctaataaata tatatgtaaa
     9121 gtacgctttt tgttgaaatt ttttaaacct ttgtttattt ttttttcttc attccgtaac
     9181 tcttctacct tctttattta ctttctaaaa tccaaataca aaacataaaa ataaataaac
     9241 acagagtaaa ttcccaaatt attccatcat taaaagatac gaggcgcgtg taagttacag
     9301 gcaagcgatc ctagtacact ctatattttt ttatgcctcg gtaatgattt tcattttttt
     9361 tttccaccta gcggatgact cttttttttt cttagcgatt ggcattatca cataatgaat
     9421 tatacattat ataaagtaat gtgatttctt cgaagaatat actaaaaaat gagcaggcaa
     9481 gataaacgaa ggcaaagatg acagagcaga aagccctagt aaagcgtatt acaaatgaaa
     9541 ccaagattca gattgcgatc tctttaaagg gtggtcccct agcgatagag cactcgatct
     9601 tcccagaaaa agaggcagaa gcagtagcag aacaggccac acaatcgcaa gtgattaacg
     9661 tccacacagg tatagggttt ctggaccata tgatacatgc tctggccaag cattccggct
     9721 ggtcgctaat cgttgagtgc attggtgact tacacataga cgaccatcac accactgaag
     9781 actgcgggat tgctctcggt caagctttta aagaggccct aggggccgtg cgtggagtaa
     9841 aaaggtttgg atcaggattt gcgcctttgg atgaggcact ttccagagcg gtggtagatc
     9901 tttcgaacag gccgtacgca gttgtcgaac ttggtttgca aagggagaaa gtaggagatc
     9961 tctcttgcga gatgatcccg cattttcttg aaagctttgc agaggctagc agaattaccc
    10021 tccacgttga ttgtctgcga ggcaagaatg atcatcaccg tagtgagagt gcgttcaagg
    10081 ctcttgcggt tgccataaga gaagccacct cgcccaatgg taccaacgat gttccctcca
    10141 ccaaaggtgt tcttatgtag ttttacacag gagtctggac ttgacgctag tgataataag
    10201 tgactgaggt atgtgctctt cttatctcct tttgtagtgt tgctcttatt ttaaacaact
    10261 ttgcggtttt ttgatgactt tgcgattttg ttgttgcttt gcagtaaatt gcaagattta
    10321 ataaaaaaac gcaaagcaat gattaaagga tgttcagaat gaaactcatg gaaacactta
    10381 accagtgcat aaacgctggt catgaaatga cgaaggctat cgccattgca cagtttaatg
    10441 atgacagccc ggaagcgagg aaaataaccc ggcgctggag aataggtgaa gcagcggatt
    10501 tagttggggt ttcttctcag gctatcagag atgccgagaa agcagggcga ctaccgcacc
    10561 cggatatgga aattcgagga cgggttgagc aacgtgttgg ttatacaatt gaacaaatta
    10621 atcatatgcg tgatgtgttt ggtacgcgat tgcgacgtgc tgaagacgta tttccaccgg
    10681 tgatcggggt tgctgcccat aaaggtggcg tttacaaaac ctcagtttct gttcatcttg
    10741 ctcaggatct ggctctgaag gggctacgtg ttttgctcgt ggaaggtaac gacccccagg
    10801 gaacagcctc aatgtatcac ggatgggtac cagatcttca tattcatgca gaagacactc
    10861 tcctgccttt ctatcttggg gaaaaggacg atgtcactta tgcaataaag cccacttgct
    10921 ggccggggct tgacattatt ccttcctgtc tggctctgca ccgtattgaa actgagttaa
    10981 tgggcaaatt tgatgaaggt aaactgccca ccgatccaca cctgatgctc cgactggcca
    11041 ttgaaactgt tgctcatgac tatgatgtca tagttattga cagcgcgcct aacctgggta
    11101 tcggcacgat taatgtcgta tgtgctgctg atgtgctgat tgttcccacg cctgctgagt
    11161 tgtttgacta cacctccgca ctgcagtttt tcgatatgct tcgtgatctg ctcaagaacg
    11221 ttgatcttaa agggttcgag cctgatgtac gtattttgct taccaaatac agcaatagca
    11281 atggctctca gtccccgtgg atggaggagc aaattcggga tgcctgggga agcatggttc
    11341 taaaaaatgt tgtacgtgaa acggatgaag ttggtaaagg tcagatccgg atgagaactg
    11401 tttttgaaca ggccattgat caacgctctt caactggtgc ctggagaaat gctctttcta
    11461 tttgggaacc tgtctgcaat gaaattttcg atcgtctgat taaaccacgc tgggagatta
    11521 gataatgaag cgtgcgcctg ttattccaaa acatacgctc aatactcaac cggttgaaga
    11581 tacttcgtta tcgacaccag ctgccccgat ggtggattcg ttaattgcgc gcgtaggagt
    11641 aatggctcgc ggtaatgcca ttactttgcc tgtatgtggt cgggatgtga agtttactct
    11701 tgaagtgctc cggggtgata gtgttgagaa gacctctcgg gtatggtcag gtaatgaacg
    11761 tgaccaggag ctgcttactg aggacgcact ggatgatctc atcccttctt ttctactgac
    11821 tggtcaacag acaccggcgt tcggtcgaag agtatctggt gtcatagaaa ttgccgatgg
    11881 gagtcgccgt cgtaaagctg ctgcacttac cgaaagtgat tatcgtgttc tggttggcga
    11941 gctggatgat gagcagatgg ctgcattatc cagattgggt aacgattatc gcccaacaag
    12001 tgcttatgaa cgtggtcagc gttatgcaag ccgattgcag aatgaatttg ctggaaatat
    12061 ttctgcgctg gctgatgcgg aaaatatttc acgtaagatt attacccgct gtatcaacac
    12121 cgccaaattg cctaaatcag ttgttgctct tttttctcac cccggtgaac tatctgcccg
    12181 gtcaggtgat gcacttcaaa aagcctttac agataaagag gaattactta agcagcaggc
    12241 atctaacctt catgagcaga aaaaagctgg ggtgatattt gaagctgaag aagttatcac
    12301 tcttttaact tctgtgctta aaacgtcatc tgcatcaaga actagtttaa gctcacgaca
    12361 tcagtttgct cctggagcga cagtattgta taagggcgat aaaatggtgc ttaacctgga
    12421 caggtctcgt gttccaactg agtgtataga gaaaattgag gccattctta aggaacttga
    12481 aaagccagca ccctgatgcg acctcgtttt agtctacgtt tatctgtctt tacttaatgt
    12541 cctttgttac aggccagaaa gcataactgg cctgaatatt ctctctgggc ccactgttcc
    12601 acttgtatcg tcggtctgat aatcagactg ggaccacggt cccactcgta tcgtcggtct
    12661 gattattagt ctgggaccac ggtcccactc gtatcgtcgg tctgattatt agtctgggac
    12721 cacggtccca ctcgtatcgt cggtctgata atcagactgg gaccacggtc ccactcgtat
    12781 cgtcggtctg attattagtc tgggaccatg gtcccactcg tatcgtcggt ctgattatta
    12841 gtctgggacc acggtcccac tcgtatcgtc ggtctgatta ttagtctgga accacggtcc
    12901 cactcgtatc gtcggtctga ttattagtct gggaccacgg tcccactcgt atcgtcggtc
    12961 tgattattag tctgggacca cgatcccact cgtgttgtcg gtctgattat cggtctggga
    13021 ccacggtccc acttgtattg tcgatcagac tatcagcgtg agactacgat tccatcaatg
    13081 cctgtcaagg gcaagtattg acatgtcgtc gtaacctgta gaacggagta acctcggtgt
    13141 gcggttgtat gcctgctgtg gattgctgct gtgtcctgct tatccacaac attttgcgca
    13201 cggttatgtg gacaaaatac ctggttaccc aggccgtgcc ggcacgttaa ccgggctgca
    13261 tccgatgcaa gtgtgtcgct gtcgacgagc tcgcgagctc ggacatgagg ttgccccgta
    13321 ttcagtgtcg ctgatttgta ttgtctgaag ttgtttttac gttaagttga tgcagatcaa
    13381 ttaatacgat acctgcgtca taattgatta tttgacgtgg tttgatggcc tccacgcacg
    13441 ttgtgatatg tagatgataa tcattatcac tttacgggtc ctttccggtg atccgacagg
    13501 ttacggggcg gcgacctcgc gggttttcgc tatttatgaa aattttccgg tttaaggcgt
    13561 ttccgttctt cttcgtcata acttaatgtt tttatttaaa ataccctctg aaaagaaagg
    13621 aaacgacagg tgctgaaagc gagctttttg gcctctgtcg tttcctttct ctgtttttgt
    13681 ccgtggaatg aacaatggaa gtccgagctc atcgctaata acttcgtata gcatacatta
    13741 tacgaagtta tattcgatgc ggccgcaagg ggttcgcgtc agcgggtgtt ggcgggtgtc
    13801 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
    13861 gtgaaatacc acacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
    13921 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
    13981 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
    14041 acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg aattcgagct
    14101 cggtacccgg ggatcctcta gagtcgacct gcaggcatgc aagcttgagt attctatagt
    14161 ctcacctaaa tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
    14221 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
    14281 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
    14341 acctgtcgtg ccagctgcat taatgaatcg gccaacgcga accccttgcg gccgcccggg
    14401 ccgtcgacca attctcatgt ttgacagctt atcatcgaat ttctgccatt catccgctta
    14461 ttatcactta ttcaggcgta gcaaccaggc gtttaagggc accaataact gccttaaaaa
    14521 aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg
    14581 acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg
    14641 tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg
    14701 gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata
    14761 ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc
    14821 gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac
    14881 gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc
    14941 tcaccgtctt tcattgccat acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg
    15001 tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta
    15061 atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa
    15121 tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc
    15181 attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat
    15241 cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc tcattttcgc
    15301 caaaagttgg cccagggctt cccggtatca acagggacac caggatttat ttattctgcg
    15361 aagtgatctt ccgtcacagg tatttattcg cgataagctc atggagcggc gtaaccgtcg
    15421 cacaggaagg acagagaaag cgcggatctg ggaagtgacg gacagaacgg tcaggacctg
    15481 gattggggag gcggttgccg ccgctgctgc tgacggtgtg acgttctctg ttccggtcac
    15541 accacatacg ttccgccatt cctatgcgat gcacatgctg tatgccggta taccgctgaa
    15601 agttctgcaa agcctgatgg gacataagtc catcagttca acggaagtct acacgaaggt
    15661 ttttgcgctg gatgtggctg cccggcaccg ggtgcagttt gcgatgccgg agtctgatgc
    15721 ggttgcgatg ctgaaacaat tatcctgaga ataaatgcct tggcctttat atggaaatgt
    15781 ggaactgagt ggatatgctg tttttgtctg ttaaacagag aagctggctg ttatccactg
    15841 agaagcgaac gaaacagtcg ggaaaatctc ccattatcgt agagatccgc attattaatc
    15901 tcaggagcct gtgtagcgtt tataggaagt agtgttctgt catgatgcct gcaagcggta
    15961 acgaaaacga tttgaatatg ccttcaggaa caatagaaat cttcgtgcgg tgttacgttg
    16021 aagtggagcg gattatgtca gcaatggaca gaacaaccta atgaacacag aaccatgatg
    16081 tggtctgtcc ttttacagcc agtagtgctc gccgcagtcg agcgacaggg cgaagcccat
    16141 cgat
//
