LOCUS       sequence-409854-        8225 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 8225)
  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 8225)
  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..8225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             422..1447
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             422..1447
                     /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"
     primer_bind     complement(3231..3250)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3350..3372
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3410..3428)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3504..3608
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3609..3677,3678..4469)
                     /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     complement(3827..3846)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4640..5228
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5129..5148
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5382..5399
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6372..6388
                     /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    6451..6467
                     /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    6590..6606
                     /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    6621..6637
                     /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    6670..6686
                     /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    6701..6717
                     /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    6732..6748
                     /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    6811..6827
                     /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    6891..6907
                     /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    6952..6968
                     /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    7033..7049
                     /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    7061..7084
                     /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    7170..7186
                     /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    7219..7235
                     /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    7250..7266
                     /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    7330..7346
                     /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    7361..7377
                     /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    7392..7408
                     /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    7440..7456
                     /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    7471..7487
                     /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    7502..7518
                     /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    7581..7597
                     /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    7612..7628
                     /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    7661..7677
                     /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    7722..7738
                     /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    7801..7817
                     /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    7832..7848
                     /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    7912..7928
                     /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    7943..7959
                     /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    8021..8037
                     /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    8052..8068
                     /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    8101..8117
                     /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 tttccaccca gcgcgggtgc attctggctc ttatatatac ttattgtcat gacagagtat
       61 attgtactgt gttgataagg gacgggtaac tgtattgaag agccgatgct tttgacatgt
      121 tagatataat atgttttatt gtaaagtcaa tacaacacac aataggataa taatgataaa
      181 gttaaaaaag tatatatagt aatagaaata tatcttatat aggaaagatt aagcagtaaa
      241 agtagcgctt acggcgtacg gagcaggaga gcaactgacc gctctcagag cccgggcaca
      301 gcaaggtctt ctgaaattca tgtttttttt ttttttactc tgcattgcag tctccgctct
      361 tatttagttt tgctttacgt aaggtctcgt tgctgccata aaataagctc tagaactagt
      421 gatgaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
      481 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
      541 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
      601 tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat
      661 tggggaattc agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
      721 gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga
      781 tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
      841 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
      901 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
      961 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
     1021 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
     1081 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
     1141 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
     1201 gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
     1261 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
     1321 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg
     1381 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
     1441 ggaatagagg cctcggagat cctaacaccg ggttgtgtgg ccaaaattgt tctgtagtcg
     1501 ctgtgagttg acacggctag tgcttatgat tttcctcgcg tgtggtgcct gtactcagcc
     1561 ctatgcctta tttgcaacac atttacgtac agcgcacaag agaagagaag atcacttgaa
     1621 gataataaat atagggttgt aggcatcttg tttaactcaa attttctcgt cttggtgtgt
     1681 cgacatgatt gaaatagtgc caccagttgt gtttgatgcg tttgttatct atgcagtatt
     1741 gcacagcaag gtcttctgaa attcatgttt tttttttttt actctgcatt gcagtctccg
     1801 ctcttattta gttttgcttt acgtaaggtc tcgttgctgc cataaaataa aattcctgca
     1861 gctacggtac ttattgccaa gaaagcacga gcatcagccg tgcctccagg tcgaatcttc
     1921 aaacgacgac gatcacgcgt cgctccagta ttccagggtt catcagatcc tacggtactt
     1981 attgccaaga aagcacgagc atcagccgtg cctccaggtc gaatcttcaa acgacgacga
     2041 tcacgcgtcg ctccagtatt ccagggttca tcagatccta cggtacttat tgccaagaaa
     2101 gcacgagcat cagccgtgcc tccaggtcga atcttcaaac gacgacgatc acgcgtcgct
     2161 ccagtattcc agggttcatc agatcctacg gtacttattg ccaagaaagc acgagcatca
     2221 gccgtgcctc caggtcgaat cttcaaacga cgacgatcac gcgtcgctcc agtattccag
     2281 ggttcatcag atcctacggt acttattgcc aagaaagcac gagcatcagc cgtgcctcca
     2341 ggtcgaatct tcaaacgacg acgatcacgc gtcgctccag tattccaggg ttcatcagat
     2401 cctacggtac ttattgccaa gaaagcacga gcatcagccg tgcctccagg tcgaatcttc
     2461 aaacgacgac gatcacgcgt cgctccagta ttccagggtt catcagatcc tacggtactt
     2521 attgccaaga aagcacgagc atcagccgtg cctccaggtc gaatcttcaa acgacgacga
     2581 tcacgcgtcg ctccagtatt ccagggttca tcagatccta cggtacttat tgccaagaaa
     2641 gcacgagcat cagccgtgcc tccaggtcga atcttcaaac gacgacgatc acgcgtcgct
     2701 ccagtattcc agggttcatc agatcctacg gtacttattg ccaagaaagc acgagcatca
     2761 gccgtgcctc caggtcgaat cttcaaacga cgacgatcac gcgtcgctcc agtattccag
     2821 ggttcatcag atcctacggt acttattgcc aagaaagcac gagcatcagc cgtgcctcca
     2881 ggtcgaatct tcaaacgacg acgatcacgc gtcgctccag tattccaggg ttcatcagat
     2941 cctacggtac ttattgccaa gaaagcacga gcatcagccg tgcctccagg tcgaatcttc
     3001 aaacgacgac gatcacgcgt cgctccagta ttccagggtt catcagatcc tacggtactt
     3061 attgccaaga aagcacgagc atcagccgtg cctccaggtc gaatcttcaa agcgccatct
     3121 ccttgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt
     3181 cctaatgcag gagtcgcata agggagagcg tcgatatggt gcactctcag tacaatctgc
     3241 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     3301 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3361 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     3421 cgcctatttt tataggttaa tgtcatgata ataatggttt cttggtttct tagacgtcag
     3481 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3541 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3601 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3661 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3721 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3781 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3841 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3901 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3961 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4021 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4081 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4141 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     4201 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     4261 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4321 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4381 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4441 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4501 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4561 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4621 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4681 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4741 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     4801 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4861 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4921 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4981 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5041 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5101 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5161 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5221 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5281 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     5341 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     5401 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     5461 gcaggaattc gagctcttta atggtcctta tacgaacgaa tattaaacaa tgcgcaggcg
     5521 cgcccatata gttggtatgt attctaattc cagactactg gcgtgggata aacatgtccc
     5581 cctgattaaa aggaaagatt ccatagccca ttagtgcaaa gataattggt acaccgtcaa
     5641 aaacacatgg gagaccacac gatacgacac tgtgaccgta gcatcaattc cgcactcgat
     5701 ataactctat cgaagaactt ccatggtaca aactggtcgc cacggtctgc tccacacgga
     5761 gatcatcgta tcatttttat cgatagcggc cgctatcgat gtatgccttg gccctgatgg
     5821 catgccaatt tcactacaac ggactatgtg gaccccgtta tcatggaaat gcgctagttg
     5881 gaggaagtta gaccgcgccg gaaaagagag gggtagagaa aatgacaact tggaagatat
     5941 ccacacacgc acggtgaaac gttagcaaca attattaggg aagcgacgct tgccctagtc
     6001 ccacgagtaa ccaagactcc aaaagccttt ctggcacaga gagcgagccg aaatggaaaa
     6061 gagaaacaat gcctgcacta acactactga gcgattcgcc tcgccgcgga ggaccgaata
     6121 ctaataacga cacttgcggt caaaaagtag aagaacaaat gctcaacgat gagtgaatca
     6181 ggttagggta gttggaaaat tatacagaat gtctttggca acacaccggt gctattatac
     6241 acaataactt ttaatgtgtc ccatattaat tacaagtgtg caacattaaa tacaagtgtg
     6301 taacattaat ttgcaagttt gcaacaatgt tctttagtgt ttaatgggtc ttgctagagt
     6361 gggtttttca attgttatcc gctcacaatg atgactttgc aattgttacc gctcgcaatt
     6421 ccgcctcgtt tgctagagtt actgtggcaa ttgttatccg ctcacaatta gcctaatgca
     6481 attgttatcc gatcacaatt tagtttcctg aattttatcc gctcacaatt gtctttacct
     6541 tgctagagct actcttcaaa ttgatccgct cacaattgcc cacccctaat tgttatccgc
     6601 tcacaatttt ccatcgttaa ttgttatccg ctcacaattt catcccccct gctagagttc
     6661 ccgctttaat tgttatccgc tcacaattta cgggtctgaa ttgttatccg ctcacaattt
     6721 catgggagga attgttatcc gctcacaatt tattactgcc tgctagagtc ccttttataa
     6781 ttgttatccg tcacaattct cttcgcctaa ttgttatccg ctcacaatta acccgcggta
     6841 attgttatcc gctcacaatt aatgctaaca tgctagagcc gctatgagaa ttgttatccg
     6901 ctcacaattt ccttccttta attgtatccg ctcacaattt cccctgtcta attgttatcc
     6961 gctcacaatt gtattttttt tgctagagcg tcactactga ttggttatcc gctcacaatt
     7021 tggcgtctgg aattgttatc cgctcacaat ttatggttat gaattgttat ccgctcacaa
     7081 ttcccctatc tctgctagag cttgtctatt aattgttatc cgttcacaat ttccgcgacc
     7141 caatagttat ccgctcacaa ttctttcaat tgttatccgc tcacaatttg tctctagttg
     7201 ctagagcatg tgtctcaatt gttatccgct cacaattcca atgtaccaat tgttatccgc
     7261 tcacaattgg cacctttcaa ttgttatccg ctcccaattt cgtcctatgt gctagagtct
     7321 tattacctat tgttatccgc tcacaattgc ccttagcgaa ttgttatccg ctcacaattt
     7381 ttctcacaca attgttatcc gctcacaatt cttccggtct gctagagctt cctacttaat
     7441 tgttatccgc tcacaatttt tttctttgta ttgttatccg ctcacaattg ctttgatcta
     7501 attgttatcc gctcacaatt ttcctttcat gctagagctc ctgtgtcaat tgttatccgc
     7561 tcataatttt agacgtcgaa ttgttatccg ctcacaattt ggtagactga attgttatcc
     7621 gctcacaatt tgccttttgc tgctagagtt ttctttttaa ttgttatccg ctcacaattt
     7681 ctttattata attgttatcc gccacaattt ctctctccta attgttatcc gctcacaatt
     7741 attgcttgct tgctagagtt cctggttcaa ttgttatccg ctccaattct tagtagctaa
     7801 ttgttatccg ctcacaattc ggcttttcta attgttatcc gctcacaatt ttcgagtcgg
     7861 tgctagagtg ttcgttcaaa ttgctatccg ctcacaattt tctaccctta attgttatcc
     7921 gctcacaatt tgaatcgctt aattgttatc cgctcacaat tgtcccttcc tgctagagtt
     7981 acttcttcaa ttgttatcgc tcacaatttt ccgtccgtaa ttgttatccg ctcacaattc
     8041 tgacccgtaa attgttatcc gctcacaatt cgtttctcca tgctagagct cttgctccaa
     8101 ttgttatccg ctcacaattt gttacgctaa tttgctatcg ctctcaattc ccattattta
     8161 attgtaatcc tccccaacca aggtccgcat tgcttttttt tttttttttt tttttttttg
     8221 ctagc
//
