LOCUS       sequence-387452-       13192 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SunTag cassette was added in the coding region in the 
            let7R-IRE-TREAT reporter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13192)
  AUTHORS   Dave P, Roth G, Griesbach E, Mateju D, Hochstoeger T, Chao JA
  TITLE     Single-molecule imaging reveals translation-dependent 
            destabilization of mRNAs.
  JOURNAL   Mol Cell. 2023 Feb 16;83(4):589-606.e6. doi: 
            10.1016/j.molcel.2023.01.013. Epub 2023 Feb 1.
  PUBMED    36731471
REFERENCE   2  (bases 1 to 13192)
  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"; doi: 
            "10.1016/j.molcel.2023.01.013"; journalName: "Mol Cell"; date: 
            "2023-02-16- 16"; volume: "83"; issue: "4"; pages: "589-606.e6"
COMMENT     Sequence Label: IRE-let7R-SunTag-TREAT
FEATURES             Location/Qualifiers
     source          1..13192
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(12..28)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12..28)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(25..47)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    36..52
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(60..66,67..84,85..90))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    105..126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(243..260)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(414..1002)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(494..513)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1173..1964,1965..2033))
                     /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     1796..1815
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2034..2138)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2206..2224
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2262..2284)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2384..2403
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2597..2619
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2611..2628
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2612..2628
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3712..3759)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     protein_bind    3806..3824
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3842..3860
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3878..3896
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3914..3932
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3950..3968
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3986..4004
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4022..4040
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             4368..4424
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             4440..4496
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             4512..4568
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             4584..4640
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             4656..4712
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             4728..4784
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             4800..4856
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             4872..4928
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             4944..5000
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5016..5072
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5088..5144
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5160..5216
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5232..5288
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5304..5360
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5376..5432
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5448..5504
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5520..5576
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5592..5648
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5664..5720
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5736..5792
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5808..5864
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             5880..5936
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             6024..6080
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             6114..7043
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="ASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAENA
                     VIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAWF
                     ELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDIA
                     LIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPREI
                     PLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKVK
                     GLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             join(7044..7046,7047..7367)
                     /codon_start=1
                     /product="destabilization domain that can be stabilized by 
                     Shield1 in the ProteoTuner(TM) system"
                     /label=DD
                     /note="L106P mutant of FKBP12"
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKPE"
     CDS             7047..7370
                     /codon_start=1
                     /product="destabilization domain that can be stabilized by 
                     Shield1 in the ProteoTuner(TM) system"
                     /label=DD-C
                     /note="FKBP12 mutant optimized for use as a C-terminal tag"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KPE"
     polyA_signal    10995..11129
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     11073..11092
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(11237..11270)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     primer_bind     complement(11323..11339)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(11355..11377)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    11783..12140
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             12205..12674
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
       61 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
      121 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
      181 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
      241 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      301 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      361 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      421 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      481 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      541 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      601 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      661 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      721 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      781 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      841 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      901 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      961 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1021 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt tgtcatgaga
     1081 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     1141 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1201 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1261 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1321 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     1381 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     1441 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     1501 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     1561 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     1621 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     1681 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     1741 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     1801 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     1861 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     1921 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1981 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     2041 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     2101 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     2161 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     2221 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     2281 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     2341 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     2401 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     2461 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     2521 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     2581 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attaattcca
     2641 agctaagtga aagactctgt ctcaaaaacc aacctgacac ctgagccgga gcgttgcctc
     2701 tcactaccat aaatatatat acacaacaca cacacacaca cacacacaca cacacacaca
     2761 cacacacaca caatcataca cataaacaca atcacacaca cacacatgca caatcacaca
     2821 cacacaatca cacacaaaca tacatacaca caatcacaca caaccataca cacatataca
     2881 caatcacaca cacaaacaca atcacacaca cacacatcaa tcaatcaatc aatcaatata
     2941 ccaaaaatga tagctcatac ctgtaatctt agatgtggga aggctaaaac agcaggctac
     3001 ccatgacttc aaggtcaccc tgggttacag agtagctaat agtctagctt gaactatagt
     3061 gtaggatctt taaaaaaaaa ttaaccaacc aaccaacaaa caaacaaaca aacaacaaca
     3121 acaacaaatg gaagacagag gacaaaaaaa aataaaagaa aaagtagaaa gccaatggag
     3181 atactatctt aaaattagaa atatttatga cacattctga atactccatg tttatttcaa
     3241 ggaagccaat gtaactttaa gtactatggc catttaatgt aaatacttaa gaaaaaaaac
     3301 caaattaatt ttgatacatg ctgcatgtga aagacccccg ctgacgggta gtcaatcact
     3361 cagaggagac cctctcaagg aacagcgaga ccacaagtcg gatgcaactg caagagggtt
     3421 tattggatac acgggtaccc gggcgactca gtcaatcgga ggactggcgc cccgagtgag
     3481 gggttgtggg ctcgagctcg gggagcagaa gcgcgcgaac agaagtcgag ctagcttgcc
     3541 aaacctacag gtggggtctt tcattccccc ctttttctgg agactaaata aaatctttta
     3601 ttttatctat ggctcgtact ctataggctt cagctggtga tattgttgag tcaaaactag
     3661 agcctggacc actgatatcc tgtctttaac aaattggact aatccggatc tgaagttcct
     3721 atactttcta gagaatagga acttcggaat aggaacttcg cggccagatc ttttgcggcc
     3781 gctttgggcc ggccgaattc tttactccct atcagtgata gagaacgtat gaagagttta
     3841 ctccctatca gtgatagaga acgtatgcag actttactcc ctatcagtga tagagaacgt
     3901 ataaggagtt tactccctat cagtgataga gaacgtatga ccagtttact ccctatcagt
     3961 gatagagaac gtatctacag tttactccct atcagtgata gagaacgtat atccagttta
     4021 ctccctatca gtgatagaga acgtataagc tttaggcgtg tacggtgggc gcctataaaa
     4081 gcagagctag tttagtgaac cgtcagatcg cctgcttcca gacgttcttc gccgagagtc
     4141 gtcggggttt cctgcttcaa cagtgcttgg acggaacccg gcgctcgttc cccaccccgg
     4201 ccggccgccc atagccagcc ctccgtcacc tcttcaccgc accctcggac tgccccaagg
     4261 cccccgccgc cgctccagcg ccgcgcagcc accgccgccg ccgccgcctc tccttagtcg
     4321 ccgccatgac cggtccgggt ggatctggag gtggaggttc tggaggagaa gaacttttga
     4381 gcaagaatta tcatcttgag aacgaagtgg ctcgtcttaa gaaaggttct ggcagtggag
     4441 aagaactgct ttcaaagaat taccacctgg aaaatgaggt agctagactg aaaaagggga
     4501 gcggaagtgg ggaggagttg ctgagcaaaa attatcattt ggagaacgaa gtagcacgac
     4561 taaagaaagg gtccggatcg ggtgaggagt tactctcgaa aaattatcat ctcgaaaacg
     4621 aagtggctcg gctaaaaaag ggcagtggtt ctggagaaga gctattatct aaaaactacc
     4681 acctcgaaaa tgaggtggca cgcttaaaaa agggaagtgg cagtggtgaa gagctactat
     4741 ccaagaatta tcatcttgag aacgaggtag cgcgtttgaa gaagggttcc ggctcaggag
     4801 aggaactgct ctcgaagaac tatcatcttg aaaatgaggt cgctcgatta aaaaagggat
     4861 cgggcagtgg tgaggaacta ctttcaaaga attaccacct cgaaaacgaa gtagctcgat
     4921 taaagaaagg ttcagggtcg ggtgaagaat tactgagtaa aaattatcat ctggaaaatg
     4981 aggtagcgag actaaaaaag gggagtggtt ctggcgaaga gttgctatcg aaaaattatc
     5041 atcttgagaa cgaagttgct aggctcaaaa agggctcagg ctcaggcgag gagttgctct
     5101 cgaaaaacta ccacttggaa aatgaggtcg cgaggttgaa aaaggggagc gggtcgggcg
     5161 aggagttatt gagcaaaaac tatcatttag agaacgaagt cgcgcgctta aagaaaggct
     5221 cgggctcggg cgaagaactc ttatcgaaga actaccacct cgaaaatgag gtcgccaggt
     5281 tgaaaaaggg cagtggcagc ggggaggaac tcttgagcaa gaactaccac ttggagaatg
     5341 aggtcgcgag attgaagaaa gggtcgggga gcggcgagga attgctcagc aagaattatc
     5401 atttggagaa cgaagtcgcc aggctcaaga aaggctcggg gtcgggggag gaattgttga
     5461 gtaaaaacta ccacttggaa aatgaagtcg ccaggctcaa aaaagggagt gggagcggcg
     5521 aagagttatt gagcaaaaat taccacttgg agaacgaagt ggcaaggctc aagaaaggga
     5581 gcggcagcgg ggaggagctc ttatcgaaga actaccactt agagaatgaa gtcgcccgct
     5641 tgaagaaagg ctcggggagc ggggaagagc tcttgagcaa gaactaccac ttggaaaatg
     5701 aggtggcgcg cttgaagaaa gggagcggga gcggggaaga gttactatct aagaattatc
     5761 atctcgagaa cgaggtggct cgactaaaga agggctccgg cagtggggag gaactcctgt
     5821 cgaagaacta tcatcttgaa aatgaggttg caagacttaa aaaggggtcc ggatcaggtg
     5881 aggaactact cagtaagaat taccacctgg aaaacgaagt agctcgatta aagaaaggtt
     5941 cagggtcggg tgaagaactg ctctcaaaag attatcattt ggaaaatgag gttgcacgtt
     6001 taaaaaaggg aagtggcagt ggtgaggaac ttctgtcgaa aaattatcat ctcgagaatg
     6061 aagtagcccg acttaaaaag ggttctggct cgggtcagcg gccgcaccgg tcagcttcca
     6121 aggtgtacga ccccgagcaa cgcaaacgca tgatcactgg gcctcagtgg tgggctcgct
     6181 gcaagcaaat gaacgtgctg gactccttca tcaactacta tgattccgag aagcacgccg
     6241 agaacgccgt gatttttctg catggtaacg ctgcctccag ctacctgtgg aggcacgtcg
     6301 tgcctcacat cgagcccgtg gctagatgca tcatccctga tctgatcgga atgggtaagt
     6361 ccggcaagag cgggaatggc tcatatcgcc tcctggatca ctacaagtac ctcaccgctt
     6421 ggttcgagct gctgaacctt ccaaagaaaa tcatctttgt gggccacgac tggggggctt
     6481 gtctggcctt tcactactcc tacgagcacc aagacaagat caaggccatc gtccatgctg
     6541 agagtgtcgt ggacgtgatc gagtcctggg acgagtggcc tgacatcgag gaggatatcg
     6601 ccctgatcaa gagcgaagag ggcgagaaaa tggtgcttga gaataacttc ttcgtcgaga
     6661 ccatgctccc aagcaagatc atgcggaaac tggagcctga ggagttcgct gcctacctgg
     6721 agccattcaa ggagaagggc gaggttagac ggcctaccct ctcctggcct cgcgagatcc
     6781 ctctcgttaa gggaggcaag cccgacgtcg tccagattgt ccgcaactac aacgcctacc
     6841 ttcgggccag cgacgatctg cctaagatgt tcatcgagtc cgaccctggg ttcttttcca
     6901 acgctattgt cgagggagct aagaagttcc ctaacaccga gttcgtgaag gtgaagggcc
     6961 tccacttcag ccaggaggac gctccagatg aaatgggtaa gtacatcaag agcttcgtgg
     7021 agcgcgtgct gaagaacgag cagatgggag tgcaggtgga aaccatctcc ccaggagacg
     7081 ggcgcacctt ccccaagcgc ggccagacct gtgtggtgca ctacaccggg atgcttgaag
     7141 atggaaagaa agtcgattcc tcccgggaca gaaacaagcc ctttaagttt atgctaggca
     7201 agcaggaggt gatccgaggc tgggaagaag gggttgccca gatgagtgtg ggtcagagag
     7261 ccaaactgac tatatctcca gattatgcct atggtgccac tgggcaccca ggcatcatcc
     7321 caccacatgc cactctcgtc ttcgatgtgg agcttctaaa accggaataa gggccaggga
     7381 agaactggct cggaaagagg accccacatg ttgtagcttc aagtttgggc cagggaagaa
     7441 ctgatcgccg tgtaaccgcg gtgaggtagt tcaataggct gtgcaggcct gaggtagttc
     7501 aataggctgt gctccgagtg aggtagttca ataggctgtc cccgcggcgc gcgcgggatc
     7561 cggatccaac ctacaaagaa cgagcagtaa gtcgacagat cctacctgca aagcaccaga
     7621 ggatatggcc tcggtgcctc agcatcttca gatgtgctgt cactgtccaa tcttcaagcc
     7681 agcagagcat atgggctcgc tggctccact gtcattacag tggacctgat cttctccaat
     7741 caaaggagca gaggatatgg cctcgctccc tccgactaca tcaggctgga agtgacatcc
     7801 atcattcatc ccagcagagc atatgggctc gctgggacca cagcatgcct cgaagtgctg
     7861 agagactatc atcaaaccga cagaggatat ggcctcgtcg gctcctactc cctcctggtg
     7921 gaatacttct ccactcttca acccagcaga gcatatgggc tcgctgggtc catcagagat
     7981 acaagatctc tcgacagatc ctacctgcaa agcaccagag gatatggcct cggtgcctca
     8041 gcatcttcag atgtgctgtc actgtccaat cttcaagcca gcagagcata tgggctcgct
     8101 ggctccactg tcattacagt ggacctgatc ttctccaatc aaaggagcag aggatatggc
     8161 ctcgctccct ccgactacat caggctggaa gtgacatcca tcattcatcc cagcagagca
     8221 tatgggctcg ctgggaccac agcatgcctc gaagtgctga gagactatca tcaaaccgac
     8281 agaggatatg gcctcgtcgg ctcctactcc ctcctggtgg aatacttctc cactcttcaa
     8341 cccagcagag catatgggct cgctgggtcc atcagagata caagatctct cgacagatcc
     8401 tacctgcaaa gcaccagagg atatggcctc ggtgcctcag catcttcaga tgtgctgtca
     8461 ctgtccaatc ttcaagccag cagagcatat gggctcgctg gctccactgt cattacagtg
     8521 gacctgatct tctccaatca aaggagcaga ggatatggcc tcgctccctc cgactacatc
     8581 aggctggaag tgacatccat cattcatccc agcagagcat atgggctcgc tgggaccaca
     8641 gcatgcctcg aagtgctgag agactatcat caaaccgaca gaggatatgg cctcgtcggc
     8701 tcctactccc tcctggtgga atacttctcc actcttcaac ccagcagagc atatgggctc
     8761 gctgggtcca tcagagatac aagatctctc gacagatcct acctgcaaag caccagagga
     8821 tatggcctcg gtgcctcagc atcttcagat gtgctgtcac tgtccaatct tcaagccagc
     8881 agagcatatg ggctcgctgg ctccactgtc attacagtgg acctgatctt ctccaatcaa
     8941 aggagcagag gatatggcct cgctccctcc gactacatca ggctggaagt gacatccatc
     9001 attcatccca gcagagcata tgggctcgct gggaccacag catgcctcga agtgctgaga
     9061 gactatcatc aaaccgacag aggatatggc ctcgtcggct cctactccct cctggtggaa
     9121 tacttctcca ctcttcaacc cagcagagca tatgggctcg ctgggtccat cagagataca
     9181 agatctctcg agccgcggtg cgtaagtctg ttaacagaat gctgcatacg ctgagtactt
     9241 cgaaatgtca tcctctgtct gacactgaac gtaatccaga cgcgtaagtc aggccggaaa
     9301 attcccgcca ccggaagttg agtagacggt gctgcctgcg actcaacccc aggaggactg
     9361 ggtgaacaaa gctgcgaagt gatccatgta agccctcaga accgtctcgg aaagaggacc
     9421 ccacatgttg tagcttcaag gcccaatgtc agaccacgcc atggcgtgcc actctgcgga
     9481 gagtgcagtc tgcgacagtg ccccaggagg actgggtgag gatcctacct acaaacggca
     9541 cgagcatcag ccgtgccgcc cagcatcttc atcactgtcc aatcttcaag cgtgcacgat
     9601 cacggcacgc tccactgtca ttacagtctt atccaatcaa actccagcag catcagcgct
     9661 ggagctccga ctacatcagg acatccatca ttcaacgctc gacgatcacg cgagcgtcca
     9721 cagcatgcct cagacactat catcaaacgg ctgcacgatc acggcagccg ctcctactgc
     9781 ctctacgtta tcgactcttc tgcgacgagc atcagccgtc gcaacatcgg acataccaga
     9841 tcctacctac aaacggcacg agcatcagcc gtgccgccca gcatcttcat cactgtccaa
     9901 tcttcaagcg tgcacgatca cggcacgctc cactgtcatt acagtcttat ccaatcaaac
     9961 tccagcagca tcagcgctgg agctccgact acatcaggac atccatcatt caacgctcga
    10021 cgatcacgcg agcgtccaca gcatgcctca gacactatca tcaaacggct gcacgatcac
    10081 ggcagccgct cctactgcct ctacgttatc gactcttctg cgacgagcat cagccgtcgc
    10141 aacatcggac ataccagatc ctacctacaa acggcacgag catcagccgt gccgcccagc
    10201 atcttcatca ctgtccaatc ttcaagcgtg cacgatcacg gcacgctcca ctgtcattac
    10261 agtcttatcc aatcaaactc cagcagcatc agcgctggag ctccgactac atcaggacat
    10321 ccatcattca acgctcgacg atcacgcgag cgtccacagc atgcctcaga cactatcatc
    10381 aaacggctgc acgatcacgg cagccgctcc tactgcctct acgttatcga ctcttctgcg
    10441 acgagcatca gccgtcgcaa catcggacat accagatcct acctacaaac ggcacgagca
    10501 tcagccgtgc cgcccagcat cttcatcact gtccaatctt caagcgtgca cgatcacggc
    10561 acgctccact gtcattacag tcttatccaa tcaaactcca gcagcatcag cgctggagct
    10621 ccgactacat caggacatcc atcattcaac gctcgacgat cacgcgagcg tccacagcat
    10681 gcctcagaca ctatcatcaa acggctgcac gatcacggca gccgctccta ctgcctctac
    10741 gttatcgact cttctgcgac gagcatcagc cgtcgcaaca tcggacatac cagatcttca
    10801 atcgatttcg cgcgggatcc agaccacctc ccctgcgagc taagctggac agccaatgac
    10861 gggtaagaga gtgacatttt tcactaacct aagacaggag ggccgtcaga gctactgcct
    10921 aatccaaaga cgggtaaaag tgataaaaat gtatcactcc aacctaagac aggcgcagct
    10981 tccgagggat ttgagatcca gacatgataa gatacattga tgagtttgga caaaccaaaa
    11041 ctagaatgca gtgaaaaaaa tgccttattt gtgaaatttg tgatgctatt gccttatttg
    11101 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
    11161 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
    11221 tggctgatta tgatctgaag ttcctatact atttgaagaa taggaacttc ggaataggaa
    11281 cttcggatct catatggcca tatgggggcg cctagagaag gagtgagggc tggataaagg
    11341 gaggatcgag gcggggtcga acgaggaggt tcaaggggga gagacggggc ggatggagga
    11401 agaggaggcg gaggcttagg gaaagggctt gacccaggga ggggggtcaa aagccaaggc
    11461 ttcccaggtc acgatgtagg ggacctggtc tgggtgtcca tgcgggccag gtgaaaagac
    11521 cttgatctta acctgggtga tgaggtctcg gttaaaggtg ccgtctcgcg gccatccgac
    11581 gttaaaggtt ggccattctg cagagcagaa ggtaacccaa cgtctcttct tgacatctac
    11641 cgactggttg tgagcgatcc gctcgacatc tttccagtga cctaaggtca aacttaaggg
    11701 agtggtaaca ggctagcccg ggcccatatt ttcagacaaa tacagaaaca cagtcagaca
    11761 gagacaacac agaacgatgc tgcagcagac aagacgcgcg gcgcggcttc ggtcccaaac
    11821 cgaaagcaaa aattcagacg gaggcgggaa ctgttttagg ttctcgtctc ctaccagaac
    11881 cacatatccc tcctctaagg ggggtgcacc aaagagtcct aaacgatcgg gattttagga
    11941 ctcaggtcgg gccacaaaaa cggcccccga agtccctggg acgtctccca gggttgcggc
    12001 cgggtgttcc gaactcgtca gttccaccac gggtccgcca gatacagagc tagttagcta
    12061 actagtaccg acgcaggcgc ataaaatcag tcatagacac tagacaatcg gacagacaca
    12121 gataagttgc tggccagctt acctcccggt ggtgggtcgg tggtccctgg gcaggggtct
    12181 cccgatcccg gacgagcccc caaatgaaag acccccgctg acgggtagtc aatcactcag
    12241 aggagaccct cccaaggaac agcgagacca caagtcggat gcaactgcaa gagggtttat
    12301 tggatacacg ggtacccggg cgactcagtc aatcggagga ctggcgcgcc gagtgagggg
    12361 ttgtgggctc ttttattgag ctcggggagc agaagcgcgc gaacagaagc gagaagcgaa
    12421 ctgattggtt agttcaaata aggcacaggg tcatttcagg tccttggggc accctggaaa
    12481 catctgatgg ttctctagaa actgctgagg gctggaccgc atctggggac catctgttct
    12541 tggccctgag ccggggcagg aactgcttac cacagatatc ctgtttggcc catattcagc
    12601 tgttccatct gttcttggcc ctgagccggg gcaggaactg cttaccacag atatcctgtt
    12661 tggcccatat tcagctgttc catctgttcc tgaccttgat ctgaactttt ctattctcag
    12721 ttatgtattt ttccatgcct tgcaaaatgg cgttacttaa gctagcttgc caaacctaca
    12781 ggtggggtct ttcacatgta tatgtcaaaa ataaaaatca actaattgac tagtaattaa
    12841 tatgactggc ataatgggaa attgatcctg acagatgcaa actggcttct cagcagcgca
    12901 tttatgttgt caactgagga aggaaacgtt aatgacagaa actctaagta atttccacgt
    12961 ttatctattt ttatttatac tagctttggt aacaggaata ttgcagcatt catgcacatt
    13021 gaaaccctta tgaaataaaa acatctgtgc atttaaaatg gaattaacat tttaaatgtt
    13081 aaaaaaagct ggcttagctt ccccccgccc cctagggcat agaacaagtc aaatgcttta
    13141 tatatttgag tttgggatgt attaggaaac tcctaagagc aaagctagct tg
//
