LOCUS       sequence-415929-        7485 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrates 50xLacO repeats and an mCherry gene upstream of the BES1 
            promoter with a hygromycin drug selection marker downstream of the 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7485)
  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 7485)
  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: pBES1-LacOx50-mCh-bla
FEATURES             Location/Qualifiers
     source          1..7485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      409..997
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     898..917
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1460..1476
                     /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    1539..1555
                     /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    1678..1694
                     /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    1709..1725
                     /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    1758..1774
                     /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    1789..1805
                     /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    1820..1836
                     /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    1899..1915
                     /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    1930..1946
                     /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    1979..1995
                     /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    2040..2056
                     /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    2089..2105
                     /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    2120..2136
                     /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    2148..2171
                     /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    2257..2273
                     /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    2306..2322
                     /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    2337..2353
                     /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    2368..2384
                     /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    2417..2433
                     /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    2448..2464
                     /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    2479..2495
                     /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    2527..2543
                     /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    2558..2574
                     /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    2589..2605
                     /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    2668..2684
                     /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    2699..2715
                     /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    2748..2764
                     /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    2809..2825
                     /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    2888..2904
                     /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    2919..2935
                     /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    2999..3015
                     /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    3030..3046
                     /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    3108..3124
                     /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    3139..3155
                     /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    3188..3204
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             3817..4215
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             4607..5317
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4752..4770)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5020..5039
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(6493..6512)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6612..6634
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6672..6690)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6758..6862
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6863..6931,6932..7485,1..238)
                     /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(7081..7100)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
       61 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
      121 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
      181 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
      241 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
      301 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
      361 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
      421 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
      481 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
      541 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
      601 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
      661 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
      721 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
      781 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
      841 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
      901 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
      961 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     1021 acggttcctg gccttttgct ggccttttgc tcacatgtct tgaacctagg tctgacgact
     1081 aaccttgatg ttaaataggt aagtttttac agatgctagg ctccaattta tagtgaagta
     1141 agcaatcgta aagttggtga ttcttccttt atgggtatga ggtaaggaat atcgacgtgc
     1201 atatcactaa agccttcttt agtacttttc acagggttag aatcactata atgctgagcg
     1261 atatttcggt tttttactaa gaaaagacac tggataatac atttgtcgca gctttgatat
     1321 gataccttat ttcccatttg tttattttta agagttatga ggcgttcttt tcacacgaga
     1381 aacacgtgat aaatgaaggt cttgcgctgt cttaattgtt aagcttatga tcgacgtctt
     1441 gctagagtgg gtttttcaat tgttatccgc tcacaatgat gactttgcaa ttgttaccgc
     1501 tcgcaattcc gcctcgtttg ctagagttac tgtggcaatt gttatccgct cacaattagc
     1561 ctaatgcaat tgttatccga tcacaattta gtttcctgaa ttttatccgc tcacaattgt
     1621 ctttaccttg ctagagctac tcttcaaatt gatccgctca caattgccca cccctaattg
     1681 ttatccgctc acaattttcc atcgttaatt gttatccgct cacaatttca tcccccctgc
     1741 tagagttccc gctttaattg ttatccgctc acaatttacg ggtctgaatt gttatccgct
     1801 cacaatttca tgggaggaat tgttatccgc tcacaattta ttactgcctg ctagagtccc
     1861 ttttataatt gttatccgtc acaattctct tcgcctaatt gttatccgct cacaattaac
     1921 ccgcggtaat tgttatccgc tcacaattaa tgctaacatg ctagagccgc tatgagaatt
     1981 gttatccgct cacaatttcc ttcctttaat tgtatccgct cacaatttcc cctgtctaat
     2041 tgttatccgc tcacaattgt attttttttg ctagagcgtc actactgatt gttatccgct
     2101 cacaatttgg cgtctggaat tgttatccgc tcacaattta tggttatgaa ttgttatccg
     2161 ctcacaattc ccctatctct gctagagctt gtctattaat tgttatccgt tcacaatttc
     2221 cgcgacccaa tagttatccg ctcacaattc tttcaattgt tatccgctca caatttgtct
     2281 ctagttgcta gagcatgtgt ctcaattgtt atccgctcac aattccaatg taccaattgt
     2341 tatccgctca caattggcac ctttcaattg ttatccgctc acaatttcgt cctatgtgct
     2401 agagtccaat tactaattgt tatccgctca caattgccct tagcgaattg ttatccgctc
     2461 acaatttttc tcacacaatt gttatccgct cacaattctt ccggtctgct agagcttcct
     2521 acttaattgt tatccgctca caattttttt ctttgtattg ttatccgctc acaattgctt
     2581 tgatctaatt gttatccgct cacaattttc ctttcatgct agagctcctg tgtcaattgt
     2641 tatccgctca taattttaga cgtcgaattg ttatccgctc acaatttggt agactgaatt
     2701 gttatccgct cacaatttgc cttttgctgc tagagttttc tttttaattg ttatccgctc
     2761 acaatttctt tattataatt gttatccgcc acaatttctc tctcctaatt gttatccgct
     2821 cacaattatt gcttgcttgc tagagttcct ggttcaattg ttatccgctc caattcttag
     2881 tagctaattg ttatccgctc acaattcggc ttttctaatt gttatccgct cacaattttc
     2941 gagtcggtgc tagagtgttc gttcaaattg ctatccgctc acaattttct acccttaatt
     3001 gttatccgct cacaatttga atcgcttaat tgttatccgc tcacaattgt cccttcctgc
     3061 tagagttact tcttcaattg ttatcgctca caattttccg tccgtaattg ttatccgctc
     3121 acaattctga cccgtaaatt gttatccgct cacaattcgt ttctccatgc tagagctctt
     3181 gctccaattg ttatccgctc acaatttgct acgctaattg ctatcgctca caattcctat
     3241 tatttaattg ttatccgcca caattaaggt ccgcttgcta gcaactgtgc tctcggagag
     3301 agagagggtg ggattttttt ctttaaataa agggagatgt attcggagga gttgcattaa
     3361 cgttgtccca tgttttacaa aaaacgagtc tgaaagaacg ccgactcaat ggccatccaa
     3421 gcggaataac cttctaaaag aatcatatcc ctattaccac accagtttat attacagggg
     3481 aggttattac agaaatctag gatatcagac tcaaggtgtg gcggacgtct cgaaccgatc
     3541 gtgagttggc ctgagcgtgc cgtcgttacc ggagtgcccg tattcgattt atgtcgactg
     3601 tgtggctgcg aaaaccctta gcgttaccac tgtgacgatt gcgtgcgatg caaagagacc
     3661 gcgtacggag caggagagca actgaccgct ctcagagccc gggcacagca aggtcttctg
     3721 aaattcatgt tttttttttt tttactctgc attgcagtct ccgctcttat ttagttttgc
     3781 tttacgtaag gtctcgttgc tgccataaaa taagctatgg ccaagccttt gtctcaagaa
     3841 gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac
     3901 tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta
     3961 tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg
     4021 gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc
     4081 ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg
     4141 aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat
     4201 gtgtgggagg gctaactcga gtaacaccgg gttgtgtggc caaatttgtt ctgtagttgc
     4261 tgtgagttga cacggctagt gcttatgatt ttcctcgcgt gtggtgcctg tactcagccc
     4321 tatgccttat ttgcaacaca tttacgtaca gcgcacaaga gaagagaaga tcacttgaag
     4381 ataataaata tagggttgta ggcatcttgt ttaactcaaa ttttctcgtc ttggtgtgtc
     4441 gacatgattg aaatagtgcc accagttgtg tttgatgcgt ttgttatcta tgcagtattg
     4501 cacagcaagg tcttctgaaa ttcatgtttt tttttttact ctgcattgca gtctccgctc
     4561 ttatttagtt ttgctttacg taaggtctcg ttgctgccat aaaataatgg tgagcaaggg
     4621 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     4681 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     4741 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     4801 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     4861 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     4921 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     4981 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     5041 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     5101 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     5161 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     5221 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     5281 ccactccacc ggcggcatgg acgagctgta caagtaggcg gccatcgaat tcctgcagcc
     5341 ctttcccccc tcaaatttcc cccctccttt taaaattttc attgctactt gaaaactttt
     5401 tgatatattt taacaccaaa accagccgag attttgtgtt ctgtgttttg taagttgact
     5461 gtctgattgt ctagaaatat tttctggcaa ctaaaatttt tttctttttt cctgtttttt
     5521 ttgtaggtag gtaggaatgg gggggggggt agttaggtag gttagttagg ttagttaggg
     5581 ggttagttag gggggttagg cttaggatta ggattagact taggcttagg attaggatta
     5641 ggattaggat tagggttaat tttttcctct ttttttttaa ctcacacctc tatcctggat
     5701 ttttaatttt tttttttagc cattcgcggc tccttttttt ttttttgcgc caatgtttaa
     5761 ttttttattg tgttttcaat ttttttgtca accatgcagc ggctgttttg ttatgcggac
     5821 cctaaccctc ctcccccccc ccgcccgcgc acctccattt ttaaaaattt ttttaccgcg
     5881 tccttcaacc agaatttttt taaatttttt aatttttttt attttccgtg gttttgaatc
     5941 ttaatttttc gaccggtacg aagagcaggg gtgcaacgga agagtgaaag gattagagga
     6001 aatggtggaa ttagcatgat cggagattgt gagggttagg aaaaacgtga agagggaaat
     6061 ggtaactgga catcgtgcgc gaggatggcg aaggcttgaa gcgtgtagag agcgtgtgta
     6121 aatggcccaa acggtgtggg tggggtaata acgcagagtg ctgaggtgtt tgacaagtgc
     6181 ccgcgagggg tgctggggag gaggaagcaa gacagtaaaa tcgtcgtgag acggtggctg
     6241 tgctcgggaa tccgtacaaa tatactgaaa attattgatg gttgatatgc gtgggaacga
     6301 cgcaggaggt acctagtaca gcgcttgttt cggcgtgggt atggtggcag gccccgtggc
     6361 cgggggactg ttgggcgcca tctccttgca ccattccttg cggcggcggt gctcaacggc
     6421 ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgaatg
     6481 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     6541 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     6601 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     6661 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     6721 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6781 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     6841 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     6901 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6961 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     7021 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     7081 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     7141 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7201 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7261 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     7321 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7381 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     7441 tggcgaacta cttactctag cttcccggca acaattaata gactg
//
