LOCUS       sequence-255889-       10021 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TPI-Wildtype NMD reporter without introns and with a 21 nt
            hairpin in 5'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10021)
  AUTHORS   Hoek TA, Khuperkar D, Lindeboom RGH, Sonneveld S, Verhagen BMP, 
            Boersma S, Vermeulen M, Tanenbaum ME
  TITLE     Single-Molecule Imaging Uncovers Rules Governing Nonsense-Mediated 
            mRNA Decay.
  JOURNAL   Mol Cell. 2019 May 23. pii: S1097-2765(19)30361-2. doi: 
            10.1016/j.molcel.2019.05.008.
  PUBMED    31155380
REFERENCE   2  (bases 1 to 10021)
  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 Cell. 
            2019 May 23. pii: S1097-2765(19)30361-2. doi: 
            10.1016/j.molcel.2019.05.008."
COMMENT     Sequence Label: HP21-TPI-WT-no_introns
FEATURES             Location/Qualifiers
     source          1..10021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        728..931
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     882..902
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    933..951
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    954..972
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     982..1006
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1145..1154
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1220..1276
                     /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             1292..1348
                     /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             1364..1420
                     /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             1436..1492
                     /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             1508..1564
                     /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             1580..1636
                     /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             1652..1708
                     /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             1724..1780
                     /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             1796..1852
                     /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             1868..1924
                     /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             1940..1996
                     /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             2012..2068
                     /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             2084..2140
                     /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             2156..2212
                     /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             2228..2284
                     /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             2300..2356
                     /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             2372..2428
                     /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             2444..2500
                     /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             2516..2572
                     /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             2588..2644
                     /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             2660..2716
                     /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             2732..2788
                     /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             2876..2932
                     /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"
     primer_bind     complement(6029..6046)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6035..6259
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6305..6733
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6392..6411)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6602..6623
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6742..6762)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6747..7076
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6927..7062
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6989..7008
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7124..7171
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7190..7564
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    7694..7815
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7731..7750)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7785..7804
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7864..7880)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7864..7880)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7877..7899)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7888..7904
                     /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(7912..7918,7919..7936,7937..7942))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7957..7978
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8095..8112)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8266..8854)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8346..8365)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9025..9816,9817..9885))
                     /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     9648..9667
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9886..9990)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaaactaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatcaag tgtatcatat
      601 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      661 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      721 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      781 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      841 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      901 tgtacggtgg gaggtctata taagcagagc tctccctatc agtgatagag atctccctat
      961 cagtgataga gatcgtcgac gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
     1021 tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc ggactctagc
     1081 gtttaaactt ccctgcggtc caccacggcc gatatcacgg ccgtggtgga ccgcagggaa
     1141 gcttgccacc atgggcgcca tcatacacgc ggccgcaatc atcggtccgg gtggatctgg
     1201 aggtggaggt tctggaggag aagaactttt gagcaagaat tatcatcttg agaacgaagt
     1261 ggctcgtctt aagaaaggtt ctggcagtgg agaagaactg ctttcaaaga attaccacct
     1321 ggaaaatgag gtagctagac tgaaaaaggg gagcggaagt ggggaggagt tgctgagcaa
     1381 aaattatcat ttggagaacg aagtagcacg actaaagaaa gggtccggat cgggtgagga
     1441 gttactctcg aaaaattatc atctcgaaaa cgaagtggct cggctaaaaa agggcagtgg
     1501 ttctggagaa gagctattat ctaaaaacta ccacctcgaa aatgaggtgg cacgcttaaa
     1561 aaagggaagt ggcagtggtg aagagctact atccaagaat tatcatcttg agaacgaggt
     1621 agcgcgtttg aagaagggtt ccggctcagg agaggaactg ctctcgaaga actatcatct
     1681 tgaaaatgag gtcgctcgat taaaaaaggg atcgggcagt ggtgaggaac tactttcaaa
     1741 gaattaccac ctcgaaaacg aagtagctcg attaaagaaa ggttcagggt cgggtgaaga
     1801 attactgagt aaaaattatc atctggaaaa tgaggtagcg agactaaaaa aggggagtgg
     1861 ttctggcgaa gagttgctat cgaaaaatta tcatcttgag aacgaagttg ctaggctcaa
     1921 aaagggctca ggctcaggcg aggagttgct ctcgaaaaac taccacttgg aaaatgaggt
     1981 cgcgaggttg aaaaagggga gcgggtcggg cgaggagtta ttgagcaaaa actatcattt
     2041 agagaacgaa gtcgcgcgct taaagaaagg ctcgggctcg ggcgaagaac tcttatcgaa
     2101 gaactaccac ctcgaaaatg aggtcgccag gttgaaaaag ggcagtggca gcggggagga
     2161 actcttgagc aagaactacc acttggagaa tgaggtcgcg agattgaaga aagggtcggg
     2221 gagcggcgag gaattgctca gcaagaatta tcatttggag aacgaagtcg ccaggctcaa
     2281 gaaaggctcg gggtcggggg aggaattgtt gagtaaaaac taccacttgg aaaatgaagt
     2341 cgccaggctc aaaaaaggga gtgggagcgg cgaagagtta ttgagcaaaa attaccactt
     2401 ggagaacgaa gtggcaaggc tcaagaaagg gagcggcagc ggggaggagc tcttatcgaa
     2461 gaactaccac ttagagaatg aagtcgcccg cttgaagaaa ggctcgggga gcggggaaga
     2521 gctcttgagc aagaactacc acttggaaaa tgaggtggcg cgcttgaaga aagggagcgg
     2581 gagcggggaa gagttactat ctaagaatta tcatctcgag aacgaggtgg ctcgactaaa
     2641 gaagggctcc ggcagtgggg aggaactcct gtcgaagaac tatcatcttg aaaatgaggt
     2701 tgcaagactt aaaaaggggt ccggatcagg tgaggaacta ctcagtaaga attaccacct
     2761 ggaaaacgaa gttgcacgtt tgaagaaagg atcaggatca ggcgaagaac tgctctcaaa
     2821 agattatcat ttggaaaatg aggttgcacg tttaaaaaag ggaagtggca gtggtgagga
     2881 acttctgtcg aaaaattatc atctcgagaa tgaagtagcc cgacttaaaa agggttctgg
     2941 ctcgggtcag cggccgcacc ggtcatctga gctgattaaa gaaaatatgc atatgaagct
     3001 gtatatggag gggacagtgg acaatcacca tttcaaatgt acatctgaag gagaggggaa
     3061 accctatgaa ggaacacaga caatgaggat caaagtggtg gaggggggcc ccctgccttt
     3121 cgcttttgac attctggcta ccagctttct gtatggaagc aagactttca tcaaccatac
     3181 ccagggaatt cctgatttct ttaaacagtc cttccctgag ggatttacat gggaacgggt
     3241 gacaacttac gaggacggcg gggtgctgac cgccactcag gatacaagcc tgcaggacgg
     3301 ctgcctgatc tataatgtga agatcagagg cgtgaacttc acttccaatg gccccgtgat
     3361 gcagaagaag accctgggct gggaggcatt cactgagact ctgtacccag ctgacggggg
     3421 actggaaggg cgcaacgata tggcactgaa actggtgggg ggaagccatc tgattgcaaa
     3481 cattaaaact acataccgct ctaaaaaacc cgcaaaaaac ctgaaaatgc caggcgtgta
     3541 ttacgtggat taccgcctgg agcgcatcaa agaggctaat aacgagacct acgtggaaca
     3601 gcacgaggtg gctgtggctc gctactgcga tctgccttct aagctgggac acaaactgaa
     3661 cgatatcgcg ccctccagga agttcttcgt tgggggaaac tggaagatga acgggcggaa
     3721 gcagagtctg ggggagctca tcggcactct gaacgcggcc aaggtgccgg ccgacaccga
     3781 ggtggtttgt gctcccccta ctgcctatat cgacttcgcc cggcagaagc tagatcccaa
     3841 gattgctgtg gctgcgcaga actgctacaa agtgactaat ggggctttta ctggggagat
     3901 cagccctggc atgatcaaag actgcggagc cacgtgggtg gtcctggggc actcagagag
     3961 aaggcatgtc tttggggagt cagatgagct gattgggcag aaagtggccc atgctctggc
     4021 agagggactc ggagtaatcg cctgcattgg ggagaagcta gatgaaaggg aagctggcat
     4081 cactgagaag gttgttttcg agcagacaaa ggtcatcgca gataacgtga aggactggag
     4141 caaggtcgtc ctggcctatg agcctgtgtg ggccattggt actggcaaga ctgcaacacc
     4201 ccaacaggcc caggaagtac acgagaagct ccgaggatgg ctgaagtcca acgtctctga
     4261 tgcggtggct cagagcaccc gtatcattta tggaggttct gtgactgggg caacctgcaa
     4321 ggagctggcc agccagcctg atgtggatgg cttccttgtg ggtggtgctt ccctcaagcc
     4381 cgaattcgtg gacatcatca atgccaaaca atgagcccca tccatcttcc ctacccttcc
     4441 tgccaagcca gggactgcta gcccggttgt acaggtaccg gatcctaagg tacctaattg
     4501 cctagaaagg agcagacgat atggcgtcgc tccctgcagg tcgactctag aaaccagcag
     4561 agcatatggg ctcgctggct gcagtattcc cgggttcatt agatcctaag gtacctaatt
     4621 gcctagaaag gagcagacga tatggcgtcg ctccctgcag gtcgactcta gaaaccagca
     4681 gagcatatgg gctcgctggc tgcagtattc ccgggttcat tagatcctaa ggtacctaat
     4741 tgcctagaaa ggagcagacg atatggcgtc gctccctgca ggtcgactct agaaaccagc
     4801 agagcatatg ggctcgctgg ctgcagtatt cccgggttca ttagatccta aggtacctaa
     4861 ttgcctagaa aggagcagac gatatggcgt cgctccctgc aggtcgactc tagaaaccag
     4921 cagagcatat gggctcgctg gctgcagtat tcccgggttc attagatcct aaggtaccta
     4981 attgcctaga aaggagcaga cgatatggcg tcgctccctg caggtcgact ctagaaacca
     5041 gcagagcata tgggctcgct ggctgcagta ttcccgggtt cattagatcc taaggtacct
     5101 aattgcctag aaaggagcag acgatatggc gtcgctccct gcaggtcgac tctagaaacc
     5161 agcagagcat atgggctcgc tggctgcagt attcccgggt tcattagatc ctaaggtacc
     5221 taattgccta gaaaggagca gacgatatgg cgtcgctccc tgcaggtcga ctctagaaac
     5281 cagcagagca tatgggctcg ctggctgcag tattcccggg ttcattagat cctaaggtac
     5341 ctaattgcct agaaaggagc agacgatatg gcgtcgctcc ctgcaggtcg actctagaaa
     5401 ccagcagagc atatgggctc gctggctgca gtattcccgg gttcattaga tcctaaggta
     5461 cctaattgcc tagaaaggag cagacgatat ggcgtcgctc cctgcaggtc gactctagaa
     5521 accagcagag catatgggct cgctggctgc agtattcccg ggttcattag atcctaaggt
     5581 acctaattgc ctagaaagga gcagacgata tggcgtcgct ccctgcaggt cgactctaga
     5641 aaccagcaga gcatatgggc tcgctggctg cagtattccc gggttcatta gatcctaagg
     5701 tacctaattg cctagaaagg agcagacgat atggcgtcgc tccctgcagg tcgactctag
     5761 aaaccagcag agcatatggg ctcgctggct gcagtattcc cgggttcatt agatcctaag
     5821 gtacctaatt gcctagaaag gagcagacga tatggcgtcg ctccctgcag gtcgactcta
     5881 gaaaccagca gagcatatgg gctcgctggc tgcagtattc ccgggttcat tagatctcgc
     5941 gaagggcgaa ttcacgaggc gcgcccggat cgcgatcgca atcttaatta actcgagtct
     6001 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     6061 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     6121 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     6181 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     6241 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     6301 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     6361 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     6421 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     6481 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     6541 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     6601 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     6661 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     6721 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     6781 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     6841 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     6901 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     6961 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     7021 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     7081 tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc
     7141 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt
     7201 gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac
     7261 cgaccggctc gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga
     7321 cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc
     7381 ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac
     7441 gaacttccgg gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg
     7501 ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga
     7561 ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     7621 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     7681 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     7741 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     7801 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     7861 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     7921 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     7981 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     8041 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     8101 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     8161 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     8221 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     8281 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     8341 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     8401 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     8461 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     8521 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     8581 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     8641 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     8701 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     8761 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     8821 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     8881 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     8941 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     9001 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     9061 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     9121 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     9181 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     9241 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     9301 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     9361 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     9421 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     9481 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     9541 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     9601 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     9661 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     9721 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     9781 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     9841 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     9901 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     9961 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
    10021 c
//
