LOCUS       sequence-51329-e       16269 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16269)
  AUTHORS   Kakirde KS, Wild J, Godiska R, Mead DA, Wiggins AG, Goodman RM, 
            Szybalski W, Liles MR
  TITLE     Gram negative shuttle BAC vector for heterologous expression of 
            metagenomic libraries.
  JOURNAL   Gene. 2011 Apr 15;475(2):57-62. Epub 2010 Nov 26.
  PUBMED    21112378
REFERENCE   2  (bases 1 to 16269)
  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: "Gene."; 
            date: "2011-04-15"; volume: "475(2)"; pages: "57-62. Epub 2010 Nov 
            26"
COMMENT     Sequence Label: pGNS-BAC
FEATURES             Location/Qualifiers
     source          1..16269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(16..21,22..38,39..44)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             233..766
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     misc_feature    967..1217
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(1220..1975)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      2066..2285
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     rep_origin      2372..3007
                     /label=oriV
                     /note="incP origin of replication"
     oriT            3712..3821
                     /note="incP origin of transfer"
     CDS             3854..4225
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     promoter        5672..5774
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             5775..6434
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5841..5860)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    6654..6675
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6690..6695,6696..6713,6714..6720)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6728..6744
                     /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)."
     primer_bind     6752..6768
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6786..6804
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     terminator      complement(6875..6902)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        6961..6979
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        7053..7498
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(7499..7585,7586..8920)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     rep_origin      9027..9615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9516..9535
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9768..9785
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(9938..9943,9944..9961,9962..9968)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9976..9992
                     /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)."
     primer_bind     10000..10016
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             10012..10296
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHATSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNS
                     EEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICSDAA"
     primer_bind     complement(10047..10063)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10056..10078)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(10272..10291)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(10403..11551)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMDFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     promoter        complement(11629..11913)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(11687..11706)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             11940..12818
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     terminator      complement(12940..12967)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(13059..13145)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    13412..13885
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     CDS             complement(13958..14929)
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             complement(14929..16104)
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
ORIGIN
        1 gcgtcaattc tcgaattgac ataagcctgt tcggttcgta aactgtaatg caagtagcgt
       61 atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg
      121 gcggttttca tggcttgtta tgactgtttt tttgtacagt ctatgcctcg ggcatccaag
      181 cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg
      241 cagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttag gtggctcaag
      301 tatgggcatc attcgcacat gtaggctcgg ccctgaccaa gtcaaatcca tgcgggctgc
      361 tcttgatctt ttcggtcgtg agttcggaga cgtagccacc tactcccaac atcagccgga
      421 ctccgattac ctcgggaact tgctccgtag taagacattc atcgcgcttg ctgccttcga
      481 ccaagaagcg gttgttggcg ctctcgcggc ttacgttctg cccaagtttg agcagccgcg
      541 tagtgagatc tatatctatg atctcgcagt ctccggcgag caccggaggc agggcattgc
      601 caccgcgctc atcaatctcc tcaagcatga ggccaacgcg cttggtgctt atgtgatcta
      661 cgtgcaagca gattacggtg acgatcccgc agtggctctc tatacaaagt tgggcatacg
      721 ggaagaagtg atgcactttg atatcgaccc aagtaccgcc acctaacaat tcgttcaagc
      781 cgagatcggc ttcccggccg acgcggctcg ccgcagccgt gtaaccgagc atagcgagcg
      841 aactggcgag gaagcaaaga agaactgttc tgtcagatag ctcttacgct cagcgcaaga
      901 agaaatatcc accgtgggaa aaactccagg tagaggtaca cacgcggata gccaattcag
      961 agtaataaac tgtgataatc aaccctcatc aatgatgacg aactaacccc cgatatcagg
     1021 tcacatgacg aagggaaaga gaaggaaatc aactgtgaca aactgccctc aaatttggct
     1081 tccttaaaaa ttacagttca aaaagtatga gaaaatccat gcaggctgaa ggaaacagca
     1141 aaactgtgac aaattaccct cagtaggtca gaacaaatgt gacgaaccac cctcaaatct
     1201 gtgacagata accctcagac tatcctgtcg tcatggaagt gatatcgcgg aaggaaaata
     1261 cgatatgagt cgtctggcgg cctttctttt tctcaatgta tgagaggcgc attggagttc
     1321 tgctgttgat ctcattaaca cagacctgca ggaagcggcg gcggaagtca ggcatacgct
     1381 ggtaactttg aggcagctgg taacgctcta tgatccagtc gattttcaga gagacgatgc
     1441 ctgagccatc cggcttacga tactgacaca gggattcgta taaacgcatg gcatacggat
     1501 tggtgatttc ttttgtttca ctaagccgaa actgcgtaaa ccggttctgt aacccgataa
     1561 agaagggaat gagatatggg ttgatatgta cactgtaaag ccctctggat ggactgtgcg
     1621 cacgtttgat aaaccaagga aaagattcat agcctttttc atcgccggca tcctcttcag
     1681 ggcgataaaa aaccacttcc ttccccgcga aactcttcaa tgcctgccgt atatccttac
     1741 tggcttccgc agaggtcaat ccgaatattt cagcatattt agcaacatgg atctcgcaga
     1801 taccgtcatg ttcctgtagg gtgccatcag attttctgat ctggtcaacg aacagataca
     1861 gcatacgttt ttgatcccgg gagagactat atgccgcctc agtgaggtcg tttgactgga
     1921 cgattcgcgg gctattttta cgtttcttgt gattgataac cgctgtttcc gccatgacag
     1981 atccatgtga agtgtgacaa gtttttagat tgtcacacta aataaaaaag agtcaataag
     2041 cagggataac tttgtgaaaa aacagcttct tctgagggca atttgtcaca gggttaaggg
     2101 caatttgtca cagacaggac tgtcatttga gggtgatttg tcacactgaa agggcaattt
     2161 gtcacaacac cttctctaga accagcatgg ataaaggcct acaaggcgct ctaaaaaaga
     2221 agatctaaaa actataaaaa aaataattat aaaaatatcc ccgtggataa gtggataacc
     2281 ccaagggaag ttttttcagg catcgtgtgt aagcagaata tataagtgct gttccctggt
     2341 gcttcctcgc tcactcgaaa ttcccctttc cgacgctcac cgggctggtt gccctcgccg
     2401 ctgggctggc ggccgtctat ggccctgcaa acgcgccaga aacgccgtcg aagccgtgtg
     2461 cgagacaccg cggccggccg ccggcgttgt ggatacctcg cggaaaactt ggccctcact
     2521 gacagatgag gggcggacgt tgacacttga ggggccgact cacccggcgc ggcgttgaca
     2581 gatgaggggc aggctcgatt tcggccggcg acgtggagct ggccagcctc gcaaatcggc
     2641 gaaaacgcct gattttacgc gagtttccca cagatgatgt ggacaagcct ggggataagt
     2701 gccctgcggt attgacactt gaggggcgcg actactgaca gatgaggggc gcgatccttg
     2761 acacttgagg ggcagagtgc tgacagatga ggggcgcacc tattgacatt tgaggggctg
     2821 tccacaggca gaaaatccag catttgcaag ggtttccgcc cgtttttcgg ccaccgctaa
     2881 cctgtctttt aacctgcttt taaaccaata tttataaacc ttgtttttaa ccagggctgc
     2941 gccctgtgcg cgtgaccgcg cacgccgaag gggggtgccc ccccttctcg aaccctcccg
     3001 gcccgctaac gcgggcctcc catcccccca ggggctgcgc ccctcggccg cgaacggcct
     3061 caccccaaaa atggcagggg atctcgaggg atcctttttg tccggtgttg ggttgaaggt
     3121 gaagccggtc ggggccgcag cgggggccgg cttttcagcc ttgcccccct gcttcggccg
     3181 ccgtggctcc ggcgtcttgg gtgccggcgc gggttccgca gccttggcct gcggtgcggg
     3241 cacatcggcg ggcttggcct tgatgtgccg cctggcgtgc gagcggaacg tctcgtagga
     3301 gaacttgacc ttccccgttt cccgcatgtg ctcccaaatg gtgacgagcg catagccgga
     3361 cgctaacgcc gcctcgacat ccgccctcac cgccaggaac gcaaccgcag cctcatcacg
     3421 ccggcgcttc ttggccgcgc gggattcaac ccactcggcc agctcgtcgg tgtagctctt
     3481 tggcatcgtc tctcgcctgt cccctcagtt cagtaatttc ctgcatttgc ctgtttccag
     3541 tcggtagata ttccacaaaa cagcagggaa gcagcgcttt tccgctgcat aaccctgctt
     3601 cggggtcatt atagcgattt tttcggtata tccatccttt ttcgcacgat atacaggatt
     3661 ttgccaaagg gttcgtgtag actttccttg gtgtacccaa cggcgtcagc cgggcaggat
     3721 aggtgaagta ggcccacccg cgagcgggtg ttccttcttc actgtccctt attcgcacct
     3781 ggcggtgctc aacgggaatc ctgctctgcg aggctggccg gctaccgccg gcgtaacaga
     3841 tgagggcaag cggatggctg atgaaaccaa gccaaccagg aagggcagcc cacctatcaa
     3901 ggtgtactgc cttccagacg aacgaagagc gattgaggaa aaggcggcgg cggccggcat
     3961 gagcctgtcg gcctacctgc tggccgtcgg ccagggctac aaaatcacgg gcgtcgtgga
     4021 ctatgagcac gtccgcgagc tggcccgcat caatggcgac ctgggccgcc tgggcggcct
     4081 gctgaaactc tggctcaccg acgacccgcg cacggcgcgg ttcggtgatg ccacgatcct
     4141 cgccctgctg gcgaagatcg aagagaagca ggacgagctt ggcaaggtca tgatgggcgt
     4201 ggtccgcccg agggcagagc catgactttt ttagccgcta aaacggccgg ggggtgcgcg
     4261 tgattgccaa gcacgtcccc atgcgctcca tcaagaagag cgacttcgcg gagctggtga
     4321 agtacatcac cgacgagcaa ggcaagaccg agcgcctggg tcacgtgcgc gtcacgaact
     4381 gcgaggcaat caccctgccc gctgtcatgg ccgaggtgat ggcgacccag cacggcaaca
     4441 cccgttccga ggccgacaag acctatcacc tgctggttag cttccgcgcg ggagagaagc
     4501 ccgacgcgga gacgttgcgc gcgattgagg accgcatctg cgctgggctt ggcttcgccg
     4561 agcatcagcg cgtcagtgcc gtgcatcacg acaccgacaa cctgcacatc catatcgcca
     4621 tcaacaagat tcacccgacc cgaaacacca tccatgagcc gtatcgggcc taccgcgccc
     4681 tcgctgacct ctgcgcgacg ctcgaacggg actacgggct tgagcgtgac aatcacgaaa
     4741 cgcggcagcg cgtttccgag aaccgcgcga acgacatgga gcggcacgcg ggcgtggaaa
     4801 gcctggtcgg ctgctcgagg gcttcgccct gtcgctcgac tgcggcgagc actactggct
     4861 gtaaaaggac agaccacatc atggttctgt gttcattagg ttgttctgtc cattgctgac
     4921 ataatccgct ccacttcaac gtaacaccgc acgaagattt ctattgttcc tgaaggcata
     4981 ttcaaatcgt tttcgttacc gcttgcaggc atcatgacag aacactactt cctataaacg
     5041 ctacacaggc tcctgagatt aataatgcgg atctctacga taatgggaga ttttcccgac
     5101 tgtttcgttc gcttctcagt ggataacagc cagcttctct gtttaacaga caaaaacagc
     5161 atatccactc agttccacat ttccatataa aggccaaggc atttattctc aggataattg
     5221 tttcagcatc gcaaccgcat cagactccgg catcgcaaac tgcacccggt gccgggcagc
     5281 cacatccagc gcaaaaacct tcgtgtagac ttccgttgaa ctgatggact tatgtcccat
     5341 caggctttgc agaactttca gcggtatacc ggcatacagc atgtgcatcg cataggaatg
     5401 gcggaacgta tgtggtgtga ccggaacaga gaacgtcaca ccgtcagcag cagcggcggc
     5461 aaccgcctcc ccaatccagg tcctgaccgt tctgtccgtc acttcccaga tccgcgcttt
     5521 ctctgtcctt cctgtgcgac ggttacgccg ctccatgagc ttatcgcgaa taaatacctg
     5581 tgacggaaga tcacttcgca gaataaataa atcctggtgt ccctgttgat accgggaagc
     5641 cctgggccaa cttttggcga aaatgagacg ttgatcggca cgtaagaggt tccaactttc
     5701 accataatga aataagatca ctaccgggcg tattttttga gttatcgaga ttttcaggag
     5761 ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat
     5821 ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga
     5881 ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt
     5941 atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg
     6001 caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc
     6061 atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt
     6121 ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta
     6181 aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt
     6241 ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat
     6301 attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt
     6361 gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc
     6421 agggcggggc gtaatttttt taaggcagtt attggtgccc ttaaacgcct ggttgctacg
     6481 cctgaataag tgataataag cggatgaatg gcagaaattc gatgataagc tgtcaaacat
     6541 gagaattggt cgacggcccg ggcggccgca aggggttcgc gttggccgat tcattaatgc
     6601 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     6661 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     6721 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     6781 aagctattta ggtgacacta tagaatactc aattctctgt ccttcctgtg cgacggttac
     6841 gccgctccat gagcttatcg ggtccactcg cgaacgcaga aaggcccacc cgaaggtgag
     6901 ccagtgtgat tacatttgcg gcctaactgt ggccagtcca gttacgctgg agtcactagt
     6961 atttaggtga cactatagaa gcggccgcga caacttgtcc acgtggaatt ctaagcttcg
     7021 gatcccactg tggtggatgc attttaaccc atcacatata cctgccgttc actattattt
     7081 agtgaaatga gatattatga tattttctga attgtgatta aaaaggcaac tttatgccca
     7141 tgcaacagaa actataaaaa atacagagaa tgaaaagaaa cagatagatt ttttagttct
     7201 ttaggcccgt agtctgcaaa tccttttatg attttctatc aaacaaaaga ggaaaataga
     7261 ccagttgcaa tccaaacgag agtctaatag aatgaggtcg aaaagtaaat cgcgcgggtt
     7321 tgttactgat aaagcaggca agacctaaaa tgtgtaaagg gcaaagtgta tactttggcg
     7381 tcacccctta catattttag gtcttttttt attgtgcgta actaacttgc catcttcaaa
     7441 caggagggct ggaagaagca gaccgctaac acagtacata aaaaaggaga catgaacgat
     7501 gaacatcaaa aagtttgcaa aacaagcaac agtattaacc tttactaccg cactgctggc
     7561 aggaggcgca actcaagcgt ttgcgaaaga aacgaaccaa aagccatata aggaaacata
     7621 cggcatttcc catattacac gccatgatat gctgcaaatc cctgaacagc aaaaaaatga
     7681 aaaatatcaa gttcctgagt tcgattcgtc cacaattaaa aatatctctt ctgcaaaagg
     7741 cctggacgtt tgggacagct ggccattaca aaacgctgac ggcactgtcg caaactatca
     7801 cggctaccac atcgtctttg cattagccgg agatcctaaa aatgcggatg acacatcgat
     7861 ttacatgttc tatcaaaaag tcggcgaaac ttctattgac agctggaaaa acgctggccg
     7921 cgtctttaaa gacagcgaca aattcgatgc aaatgattct atcctaaaag accaaacaca
     7981 agaatggtca ggttcagcca catttacatc tgacggaaaa atccgtttat tctacactga
     8041 tttctccggt aaacattacg gcaaacaaac actgacaact gcacaagtta acgtatcagc
     8101 atcagacagc tctttgaaca tcaacggtgt agaggattat aaatcaatct ttgacggtga
     8161 cggaaaaacg tatcaaaatg tacagcagtt catcgatgaa ggcaactaca gctcaggcga
     8221 caaccatacg ctgagagatc ctcactacgt agaagataaa ggccacaaat acttagtatt
     8281 tgaagcaaac actggaactg aagatggcta ccaaggcgaa gaatctttat ttaacaaagc
     8341 atactatggc aaaagcacat cattcttccg tcaagaaagt caaaaacttc tgcaaagcga
     8401 taaaaaacgc acggctgagt tagcaaacgg cgctctcggt atgattgagc taaacgatga
     8461 ttacacactg aaaaaagtga tgaaaccgct gattgcatct aacacagtaa cagatgaaat
     8521 tgaacgcgcg aacgtcttta aaatgaacgg caaatggtac ctgttcactg actcccgcgg
     8581 atcaaaaatg acgattgacg gcattacgtc taacgatatt tacatgcttg gttatgtttc
     8641 taattcttta actggcccat acaagccgct gaacaaaact ggccttgtgt taaaaatgga
     8701 tcttgatcct aacgatgtaa cctttactta ctcacacttc gctgtacctc aagcgaaagg
     8761 aaacaatgtc gtgattacaa gctatatgac aaacagagga ttctacgcag acaaacaatc
     8821 aacgtttgcg ccaagtttcc tgctgaacat caaaggcaag aaaacatctg ttgtcaaaga
     8881 cagcatcctt gaacaaggac aattaacagt taacaaataa aaacgcaaaa gaaaatgccg
     8941 atatcctatt ggcattttct tttatttctt atcaacataa aggtgaatcc gacgtctgtg
     9001 ctggtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     9061 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     9121 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     9181 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     9241 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     9301 gactcaagac gatagttact ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     9361 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     9421 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     9481 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     9541 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     9601 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     9661 ccttttgctc acatgttctt tcctgcgtat cccctgattc tgtggataac cgtattaccg
     9721 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     9781 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     9841 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     9901 ttaatgtggg ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     9961 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
    10021 attacgccaa gtttgcatgc gacttcactg gccgtcgttt tacaacgtcg tgactgggaa
    10081 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
    10141 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
    10201 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
    10261 tgcactctca gtacaatctg ctctgatgcc gcatagccat cacattggga tcctaagctt
    10321 gatatcgcat gcctgcagcc cgggagatct actagttcta ggtggctgct gaacccccag
    10381 ccggaactga ccccacaagg ccctagcgtt tgcaatgcac caggtcatca ttgacccagg
    10441 cgtgttccac caggccgctg cctcgcaact cttcgcaggc ttcgccgacc tgctcgcgcc
    10501 acttcttcac gcgggtggaa tccgatccgc acatgaggcg gaaggtttcc agcttgagcg
    10561 ggtacggctc ccggtgcgag ctgaaatagt cgaacatccg tcgggccgtc ggcgacagct
    10621 tgcggtactt ctcccatatg aatttcgtgt agtggtcgcc agcaaacagc acgacgattt
    10681 cctcgtcgat caggacctgg caacgggacg ttttcttgcc acggtccagg acgcggaagc
    10741 ggtgcagcag cgacaccgat tccaggtgcc caacgcggtc ggacgtgaag tccatcgccg
    10801 tcgcctgtag gcgcgacagg cattcctcgg ccttcgtgta ataccggcca ttgatcgacc
    10861 agcccaggtc ctggcaaagc tcgtagaacg tgaaggtgat cggctcgccg ataggggtgc
    10921 gcttcgcgta ctccaacacc tgctgccaca ccagttcgtc atcgtcggcc cgcagctcga
    10981 cgccggtgta ggtgatcttc acgtccttgt tgacgtggaa aatgaccttg ttttgcagcg
    11041 cctcgcgcgg gattttcttg ttgcgcgtgg tgaacagggc agagcgggcc gtgtcgtttg
    11101 gcatcgctcg catcgtgtcc ggccacggcg caatatcgaa caaggaaagc tgcatttcct
    11161 tgatctgctg cttcgtgtgt ttcagcaacg cggcctgctt ggcctcgctg acctgttttg
    11221 ccaggtcctc gccggcggtt tttcgcttct tggtcgtcat agttcctcgc gtgtcgatgg
    11281 tcatcgactt cgccaaacct gccgcctcct gttcgagacg acgcgaacgc tccacggcgg
    11341 ccgatggcgc gggcagggca gggggagcca gttgcacgct gtcgcgctcg atcttggccg
    11401 tagcttgctg gaccatcgag ccgacggact ggaaggtttc gcggggcgca cgcatgacgg
    11461 tgcggcttgc gatggtttcg gcatcctcgg cggaaaaccc cgcgtcgatc agttcttgcc
    11521 tgtatgcctt ccggtcaaac gtccgattca ttcaccctcc ttgcgggatt gccccggaat
    11581 ttagagccat ggtgaattcc tcctttcact ccatccaaaa aaacgggtat ggagaaacag
    11641 tagagagttg cgataaaaag cgtcaggtag gatccgctaa tcttatggat aaaaatgcta
    11701 tggcatagca aagtgtgacg ccgtgcaaat aatcaatgtg gacttttctg ccgtgattat
    11761 agacactttt gttacgcgtt tttgtcatgg ctttggtccc gctttgttac agaatgcttt
    11821 taataagcgg ggttaccggt tgggttagcg agaagagcca gtaaaagacg cagtgacggc
    11881 aatgtctgat gcaatatgga caattggttt cttctctgaa tggtgggagt atgaaaagta
    11941 tggctgaagc gcaaaatgat cccctgctgc cgggatactc gtttaacgcc catctggtgg
    12001 cgggtttaac gccgattgag gccaacggtt atctcgattt ttttatcgac cgaccgctgg
    12061 gaatgaaagg ttatattctc aatctcacca ttcgcggtca gggggtggtg aaaaatcagg
    12121 gacgagaatt tgtctgccga ccgggtgata ttttgctgtt cccgccagga gagattcatc
    12181 actacggtcg tcatccggag gctcgcgaat ggtatcacca gtgggtttac tttcgtccgc
    12241 gcgcctactg gcatgaatgg cttaactggc cgtcaatatt tgccaatacg ggtttctttc
    12301 gcccggatga agcgcaccag ccgcatttca gcgacctgtt tgggcaaatc attaacgccg
    12361 ggcaagggga agggcgctat tcggagctgc tggcgataaa tctgcttgag caattgttac
    12421 tgcggcgcat ggaagcgatt aacgagtcgc tccatccacc gatggataat cgggtacgcg
    12481 aggcttgtca gtacatcagc gatcacctgg cagacagcaa ttttgatatc gccagcgtcg
    12541 cacagcatgt ttgcttgtcg ccgtcgcgtc tgtcacatct tttccgccag cagttaggga
    12601 ttagcgtctt aagctggcgc gaggaccaac gcattagtca ggcgaagctg cttttgagca
    12661 ctacccggat gcctatcgcc accgtcggtc gcaatgttgg ttttgacgat caactctatt
    12721 tctcgcgagt atttaaaaaa tgcaccgggg ccagcccgag cgagtttcgt gccggttgtg
    12781 aagaaaaagt gaatgatgta gccgtcaagt tgtcataatt ggtaacgaat cagacaattg
    12841 acggcttgac ggagtagcat agggtttgca gaatccctgc ttcgtccatt tgacaggcca
    12901 cattatgcaa gcattgcgga gcttggcact ggccacgcaa aaaggccatc cgtcaggatg
    12961 gccttctgct taatttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg
    13021 ggccgttgct tcgcaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg
    13081 ttcaccgaca aacaacagat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt
    13141 atttgatgcc tggcagttcc ctactctcgc atggggagac cccacactac catcgggggc
    13201 catcgaacgt gccggcacgg cctgggtaac caggtatttt gtccacataa ccgtgcgcaa
    13261 aatgttgtgg ataagcagga cacagcagca atccacagca ggcatacaac cgcacaccga
    13321 ggttactccg ttctacaggt tacgacgaca tgtcaatact tgcccttgac aggcattgat
    13381 ggaatcgtag tctcacgctg atagtctgat cgacaataca agtgggaccg tggtcccaga
    13441 ccgataatca gaccgacaac acgagtggga tcgtggtccc agactaataa tcagaccgac
    13501 gatacgagtg ggaccgtggt cccagactaa taatcagacc gacgatacga gtgggaccgt
    13561 ggttccagac taataatcag accgacgata cgagtgggac cgtggtccca gactaataat
    13621 cagaccgacg atacgagtgg gaccatggtc ccagactaat aatcagaccg acgatacgag
    13681 tgggaccgtg gtcccagtct gattatcaga ccgacgatac gagtgggacc gtggtcccag
    13741 actaataatc agaccgacga tacgagtggg accgtggtcc cagactaata atcagaccga
    13801 cgatacgagt gggaccgtgg tcccagtctg attatcagac cgacgataca agtggaacag
    13861 tgggcccaga gagaatattc aggccagtta tgctttctgg cctgtaacaa aggacattaa
    13921 gtaaagacag ataaacgtag actaaaacgt ggtcgcatca gggtgctggc ttttcaagtt
    13981 ccttaagaat ggcctcaatt ttctctatac actcagttgg aacacgagac ctgtccaggt
    14041 taagcaccat tttatcgccc ttatacaata ctgtcgctcc aggagcaaac tgatgtcgtg
    14101 agcttaaact agttcttgat gcagatgacg ttttaagcac agaagttaaa agagtgataa
    14161 cttcttcagc ttcaaatatc accccagctt ttttctgctc atgaaggtta gatgcctgct
    14221 gcttaagtaa ttcctcttta tctgtaaagg ctttttgaag tgcatcacct gaccgggcag
    14281 atagttcacc ggggtgagaa aaaagagcaa caactgattt aggcaatttg gcggtgttga
    14341 tacagcgggt aataatctta cgtgaaatat tttccgcatc agccagcgca gaaatatttc
    14401 cagcaaattc attctgcaat cggcttgcat aacgctgacc acgttcataa gcacttgttg
    14461 ggcgataatc gttacccaat ctggataatg cagccatctg ctcatcatcc agctcgccaa
    14521 ccagaacacg ataatcactt tcggtaagtg cagcagcttt acgacggcga ctcccatcgg
    14581 caatttctat gacaccagat actcttcgac cgaacgccgg tgtctgttga ccagtcagta
    14641 gaaaagaagg gatgagatca tccagtgcgt cctcagtaag cagctcctgg tcacgttcat
    14701 tacctgacca tacccgagag gtcttctcaa cactatcacc ccggagcact tcaagagtaa
    14761 acttcacatc ccgaccacat acaggcaaag taatggcatt accgcgagcc attactccta
    14821 cgcgcgcaat taacgaatcc accatcgggg cagctggtgt cgataacgaa gtatcttcaa
    14881 ccggttgagt attgagcgta tgttttggaa taacaggcgc acgcttcatt atctaatctc
    14941 ccagcgtggt ttaatcagac gatcgaaaat ttcattgcag acaggttccc aaatagaaag
    15001 agcatttctc caggcaccag ttgaagagcg ttgatcaatg gcctgttcaa aaacagttct
    15061 catccggatc tgacctttac caacttcatc cgtttcacgt acaacatttt ttagaaccat
    15121 gcttccccag gcatcccgaa tttgctcctc catccacggg gactgagagc cattactatt
    15181 gctgtatttg gtaagcaaaa tacgtacatc aggctcgaac cctttaagat caacgttctt
    15241 gagcagatca cgaagcatat cgaaaaactg cagtgcggag gtgtagtcaa acaactcagc
    15301 aggcgtggga acaatcagca catcagcagc acatacgaca ttaatcgtgc cgatacccag
    15361 gttaggcgcg ctgtcaataa ctatgacatc atagtcatga gcaacagttt caatggccag
    15421 tcggagcatc aggtgtggat cggtgggcag tttaccttca tcaaatttgc ccattaactc
    15481 agtttcaata cggtgcagag ccagacagga aggaataatg tcaagccccg gccagcaagt
    15541 gggctttatt gcataagtga catcgtcctt ttccccaaga tagaaaggca ggagagtgtc
    15601 ttctgcatga atatgaagat ctggtaccca tccgtgatac attgaggctg ttccctgggg
    15661 gtcgttacct tccacgagca aaacacgtag ccccttcaga gccagatcct gagcaagatg
    15721 aacagaaact gaggttttgt aaacgccacc tttatgggca gcaaccccga tcaccggtgg
    15781 aaatacgtct tcagcacgtc gcaatcgcgt accaaacaca tcacgcatat gattaatttg
    15841 ttcaattgta taaccaacac gttgctcaac ccgtcctcga atttccatat ccgggtgcgg
    15901 tagtcgccct gctttctcgg catctctgat agcctgagaa gaaaccccaa ctaaatccgc
    15961 tgcttcacct attctccagc gccgggttat tttcctcgct tccgggctgt catcattaaa
    16021 ctgtgcaatg gcgatagcct tcgtcatttc atgaccagcg tttatgcact ggttaagtgt
    16081 ttccatgagt ttcattctga acatccttta atcattgctt tgcgtttttt tattaaatct
    16141 tgcaatttac tgcaaagcaa caacaaaatc gcaaagtcat caaaaaaccg caaagttgtt
    16201 taaaataaga gcaacactac aaaaggagat aagaagagca catacctcag tcacttatta
    16261 tcactagcc
//
