LOCUS       sequence-415840-        7731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrates 50xLacO repeats and 24xMS2 repeats with a hygromycin drug
            selection marker into the rDNA spacer..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7731)
  AUTHORS   Budzak J, Goodwin I, Tiengwe C, Rudenko G
  TITLE     Imaging of genomic loci in Trypanosoma brucei using an optimised 
            LacO-LacI system.
  JOURNAL   Mol Biochem Parasitol. 2023 Dec;256:111598. doi: 
            10.1016/j.molbiopara.2023.111598. Epub 2023 Nov 3.
  PUBMED    37923299
REFERENCE   2  (bases 1 to 7731)
  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: "Mol Biochem
            Parasitol."; date: "2023-12"; volume: "256:111598. doi"; pages: " 
            10.1016/j.molbiopara.2023.111598. Epub 2023 Nov 3"
COMMENT     Sequence Label: prDNA spacer-LacOx50-20xMS2-Hyg
FEATURES             Location/Qualifiers
     source          1..7731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(896..913)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1067..1655)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1147..1166)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1826..2617,2618..2686))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2449..2468
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2687..2791)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2859..2877
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2915..2937)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3037..3056
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3266..3285
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4386..5411)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(4386..5411)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     protein_bind    5911..5927
                     /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)."
     protein_bind    5960..5976
                     /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)."
     protein_bind    5991..6007
                     /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)."
     protein_bind    6069..6085
                     /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)."
     protein_bind    6100..6116
                     /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)."
     protein_bind    6180..6196
                     /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)."
     protein_bind    6211..6227
                     /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)."
     protein_bind    6290..6306
                     /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)."
     protein_bind    6351..6367
                     /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)."
     protein_bind    6400..6416
                     /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)."
     protein_bind    6431..6447
                     /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)."
     protein_bind    6510..6526
                     /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)."
     protein_bind    6541..6557
                     /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)."
     protein_bind    6572..6588
                     /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)."
     protein_bind    6620..6636
                     /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)."
     protein_bind    6651..6667
                     /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)."
     protein_bind    6682..6698
                     /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)."
     protein_bind    6731..6747
                     /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)."
     protein_bind    6762..6778
                     /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)."
     protein_bind    6793..6809
                     /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)."
     protein_bind    6842..6858
                     /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)."
     protein_bind    6944..6967
                     /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)."
     protein_bind    6979..6995
                     /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)."
     protein_bind    7010..7026
                     /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)."
     protein_bind    7059..7075
                     /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)."
     protein_bind    7120..7136
                     /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)."
     protein_bind    7169..7185
                     /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)."
     protein_bind    7200..7216
                     /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)."
     protein_bind    7279..7295
                     /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)."
     protein_bind    7310..7326
                     /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)."
     protein_bind    7341..7357
                     /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)."
     protein_bind    7390..7406
                     /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)."
     protein_bind    7421..7437
                     /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)."
     protein_bind    7560..7576
                     /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)."
     protein_bind    7639..7655
                     /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)."
ORIGIN
        1 ttgtatttaa tgttgcacac ttgtaattaa tatgggacac attaaaagtt attgtgtata
       61 atagcaccgg tgtgttgcca aagacattct gtataatttt ccaactaccc taacctgatt
      121 cactcatcgt tgagcatttg ttcttctact ttttgaccgc aagtgtcgtt attagtattc
      181 ggtcctccgc ggcgaggcga atcgctcagt agtgttagtg caggcattgt ttctcttttc
      241 catttcggct cgctctctgt gccagaaagg cttttggagt cttggttact cgtgggacta
      301 gggcaagcgt cgcttcccta ataattgttg ctaacgtttc accgtgcgtg tgtggatatc
      361 ttccaagttg tcattttctc tacccctctc ttttccggcg cggtctaact tcctccaact
      421 agcgcatttc catgataacg gggtccacat agtccgttgt agtgaaattg gcatgccatc
      481 agggccaagg catacatcga tagcggccgc tatcgataaa aatgatacga tgatctccgt
      541 gtggagcaga ccgtggcgac cagtttgtac catggaagtt cttcgataga gttatatcga
      601 gtgcggaatt gatgctacgg tcacagtgtc gtatcgtgtg gtctcccatg tgtttttgac
      661 ggtgtaccaa ttatctttgc actaatgggc tatggaatct ttccttttaa tcagggggac
      721 atgtttatcc cacgccagta gtctggaatt agaatacata ccaactatat gggcgcgcct
      781 gcgcattgtt taatattcgt tcgtataagg accattaaag agctcgaatt ctgcattaat
      841 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
      901 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
      961 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1021 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1081 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1141 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1201 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     1261 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1321 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1381 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     1441 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     1501 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     1561 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     1621 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     1681 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     1741 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     1801 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     1861 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     1921 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     1981 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     2041 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     2101 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     2161 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     2221 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     2281 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     2341 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     2401 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     2461 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     2521 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     2581 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     2641 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     2701 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     2761 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     2821 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     2881 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     2941 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     3001 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     3061 gagtgcacca ttcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt
     3121 aggttgaggc cgttgagcac cgccgccgca aggaatggtg caaggagatg gcgcccaaca
     3181 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
     3241 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
     3301 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatctgg ctaggaatac
     3361 tgcatagata acaaacgcat caaacacaac tggtggcact atttcaatca tgtcgagatc
     3421 ctacggtact tattgccaag aaagcacgag catcagccgt gcctccaggt cgaatcttca
     3481 aacgacgacg atcacgcgtc gctccagtat tccagggttc atcagatcct acggtactta
     3541 ttgccaagaa agcacgagca tcagccgtgc ctccaggtcg aatcttcaaa cgacgacgat
     3601 cacgcgtcgc tccagtattc cagggttcat cagatcctac ggtacttatt gccaagaaag
     3661 cacgagcatc agccgtgcct ccaggtcgaa tcttcaaacg acgacgatca cgcgtcgctc
     3721 cagtattcca gggttcatca gatcctacgg tacttattgc caagaaagca cgagcatcag
     3781 ccgtgcctcc aggtcgaatc ttcaaacgac gacgatcacg cgtcgctcca gtattccagg
     3841 gttcatcaga tcctacggta cttattgcca agaaagcacg agcatcagcc gtgcctccag
     3901 gtcgaatctt caaacgacga cgatcacgcg tcgctccagt attccagggt tcatcagatc
     3961 gggctgcagg aattagcttt attttatggc agcaacgaga ccttacgtaa agcaaaacta
     4021 aataagagcg gagactgcaa tgcagagtaa aaaaaaaaaa acatgaattt cagaagacct
     4081 tgctgtgcaa tactgcatag ataacaaacg catcaaacac aactggtggc actatttcaa
     4141 tcatgtcgat cgacacacca agacgagaaa atttgagtta aacaagatgc ctacaaccct
     4201 atatttatta tcttcaagtg atcttctctt ctcttgtgcg ctgtacgtaa atgtgttgca
     4261 aataaggcat agggctgagt acaggcacca cacgcgagga aaatcataag cactagccgt
     4321 gtcaactcac agcgactaca gaacaatttt ggccacacaa cccggtgtta ggatctccga
     4381 ggcctctatt cctttgccct cggacgagtg ctggggcgtc ggtttccact atcggcgagt
     4441 acttctacac agccatcggt ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg
     4501 acagtcccgg ctccggatcg gacgattgcg tcgcatcgac cctgcgccca agctgcatca
     4561 tcgaaattgc cgtcaaccaa gctctgatag agttggtcaa gaccaatgcg gagcatatac
     4621 gcccggagcc gcggcgatcc tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc
     4681 tgctccatac aagccaacca cggcctccag aagaagatgt tggcgacctc gtattgggaa
     4741 tccccgaaca tcgcctcgct ccagtcaatg accgctgtta tgcggccatt gtccgtcagg
     4801 acattgttgg agccgaaatc cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc
     4861 caaagcatca gctcatcgag agcctgcgcg acggacgcac tgacggtgtc gtccatcaca
     4921 gtttgccagt gatacacatg gggatcagca atcgcgcata tgaaatcacg ccatgtagtg
     4981 tattgaccga ttccttgcgg tccgaatggg ccgaacccgc tcgtctggct aagatcggcc
     5041 gcagcgatcg catccatggc ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca
     5101 ggcaggtctt gcaacgtgac accctgtgca cggcgggaga tgcaataggt caggctctcg
     5161 ctgaattccc caatgtcaag cacttccgga atcgggagcg cggccgatgc aaagtgccga
     5221 taaacataac gatctttgta gaaaccatcg gcgcagctat ttacccgcag gacatatcca
     5281 cgccctccta catcgaagct gaaagcacga gattcttcgc cctccgagag ctgcatcagg
     5341 tcggagacgc tgtcgaactt ttcgatcaga aacttctcga cagacgtcgc ggtgagttca
     5401 ggctttttca tcactagttc tagagcttat tttatggcag caacgagacc ttacgtaaag
     5461 caaaactaaa taagagcgga gactgcaatg cagagtaaaa aaaaaaaaaa catgaatttc
     5521 agaagacctt gctgtgcccg ggctctgaga gcggtcagtt gctctcctgc tccgtacgcc
     5581 gtaagcgcta cttttactgc ttaatctttc ctatataaga tatatttcta ttactatata
     5641 tactttttta actttatcat tattatccta ttgtgtgttg tattgacttt acaataaaac
     5701 atattatatc taacatgtca aaagcatcgg ctcttcaata cagttacccg tcccttatca
     5761 acacagtaca atatactctg tcatgacaat aagtatatat aagagccaga atgcacccgc
     5821 gctgggtgga aagctagcaa gcggacctta attgtggcgg ataacaatta aataatagga
     5881 attgtgagcg atagcaatta gcgtagcaaa ttgtgagcgg ataacaattg gagcaagagc
     5941 tctagcatgg agaaacgaat tgtgagcgga taacaattta cgggtcagaa ttgtgagcgg
     6001 ataacaatta cggacggaaa attgtgagcg ataacaattg aagaagtaac tctagcagga
     6061 agggacaatt gtgagcggat aacaattaag cgattcaaat tgtgagcgga taacaattaa
     6121 gggtagaaaa ttgtgagcgg atagcaattt gaacgaacac tctagcaccg actcgaaaat
     6181 tgtgagcgga taacaattag aaaagccgaa ttgtgagcgg ataacaatta gctactaaga
     6241 attggagcgg ataacaattg aaccaggaac tctagcaagc aagcaataat tgtgagcgga
     6301 taacaattag gagagagaaa ttgtggcgga taacaattat aataaagaaa ttgtgagcgg
     6361 ataacaatta aaaagaaaac tctagcagca aaaggcaaat tgtgagcgga taacaattca
     6421 gtctaccaaa ttgtgagcgg ataacaattc gacgtctaaa attatgagcg gataacaatt
     6481 gacacaggag ctctagcatg aaaggaaaat tgtgagcgga taacaattag atcaaagcaa
     6541 ttgtgagcgg ataacaatac aaagaaaaaa attgtgagcg gataacaatt aagtaggaag
     6601 ctctagcaga ccggaagaat tgtgagcgga taacaattgt gtgagaaaaa ttgtgagcgg
     6661 ataacaattc gctaagggca attgtgagcg gataacaatt agtaattgga ctctagcaca
     6721 taggacgaaa ttgtgagcgg ataacaattg aaaggtgcca attgtgagcg gataacaatt
     6781 ggtacattgg aattgtgagc ggataacaat tgagacacat gctctagcaa ctagagacaa
     6841 attgtgagcg gataacaatt gaaagaattg tgagcggata actattgggt cgcggaaatt
     6901 gtgaacggat aacaattaat agacaagctc tagcagagat aggggaattg tgagcggata
     6961 acaattcata accataaatt gtgagcggat aacaattcca gacgccaaat tgtgagcgga
     7021 taacaatcag tagtgacgct ctagcaaaaa aaatacaatt gtgagcggat aacaattaga
     7081 caggggaaat tgtgagcgga tacaattaaa ggaaggaaat tgtgagcgga taacaattct
     7141 catagcggct ctagcatgtt agcattaatt gtgagcggat aacaattacc gcgggttaat
     7201 tgtgagcgga taacaattag gcgaagagaa ttgtgacgga taacaattat aaaagggact
     7261 ctagcaggca gtaataaatt gtgagcggat aacaattcct cccatgaaat tgtgagcgga
     7321 taacaattca gacccgtaaa ttgtgagcgg ataacaatta aagcgggaac tctagcaggg
     7381 gggatgaaat tgtgagcgga taacaattaa cgatggaaaa ttgtgagcgg ataacaatta
     7441 ggggtgggca attgtgagcg gatcaatttg aagagtagct ctagcaaggt aaagacaatt
     7501 gtgagcggat aaaattcagg aaactaaatt gtgatcggat aacaattgca ttaggctaat
     7561 tgtgagcgga taacaattgc cacagtaact ctagcaaacg aggcggaatt gcgagcggta
     7621 acaattgcaa agtcatcatt gtgagcggat aacaattgaa aaacccactc tagcaagacc
     7681 cattaaacac taaagaacat tgttgcaaac ttgcaaatta atgttacaca c
//
