LOCUS       sequence-255918-       11838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TPI-PTC1 NMD reporter (It has a Premature termination 
            codon (PTC) at First Amino acid) with a 21 nt hairpin in 5'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11838)
  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 11838)
  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-PTC1
FEATURES             Location/Qualifiers
     source          1..11838
                     /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(7846..7863)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7852..8076
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8122..8550
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8209..8228)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8419..8440
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8559..8579)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8564..8893
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8744..8879
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8806..8825
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8941..8988
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9007..9381
                     /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    9511..9632
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9548..9567)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9602..9621
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9681..9697)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9681..9697)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9694..9716)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9705..9721
                     /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(9729..9735,9736..9753,9754..9759))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9774..9795
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9912..9929)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10083..10671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10163..10182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10842..11633,11634..11702))
                     /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     11465..11484
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11703..11807)
                     /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 cgatatctaa ccctccagga agttcttcgt tgggggaaac tggaagatga acgggcggaa
     3721 gcagagtctg ggggagctca tcggcactct gaacgcggcc aaggtgccgg ccgacaccgg
     3781 taagccctcg ccgaggaggg gtctggccgg gccggggccg gggccggggc aggagtggca
     3841 gcgccctctc ccgaggcccc gaggccccga ggccggtatc cgcgcggacc tgatgcaggg
     3901 ctgtgggacg agggccgctg gggtccgggc aggggcctcg cagccgcagc cccgtcggtg
     3961 cgtcgagggg gcagggcgga gcacatgatg ccccttggac tatggggcag gtaaggacgt
     4021 tttcttccct tcggcaacct gaacgactcc cgccttccac ggaagggacc gagcccgtgc
     4081 caaacaggct gagcgatttg ggagtgagga gccatcctac cgctttcccc aacctggaaa
     4141 cagcaaagcg caaggcctct gagtcagtta ggtctctgcc acccacgggc aaaggatgct
     4201 ctcctccatc ctccttcctc cctccaccga aatcggagag ccgcgggcct gatccaaaga
     4261 ggcatcccct tctcgttcat tccccagagg cctcaataca aaccccagga gttggcccct
     4321 ctccttttgc tacaaatcct tgccttgcaa aggggaggtg aggatgggct attttagaag
     4381 ggaagcaggg ttgctccctg gagaatgctg agtctgtgag gtgcctatgc cgagaatagg
     4441 tcgaggaaat tggagcccca gctgttaaaa gagcagaggg cagggtgagg gccgtggcct
     4501 ctcaggggta tctggaaggc tcttcgagtt gagtgcagac ccagcctggg ctggaaaatg
     4561 gacaaaggtc atcttgctgg ggtgaaaagg gggagagcag aaccaagaag aagagggtga
     4621 gggctggggg gctccagggc actggttagg aattgtgggg aatgaaggct ttctttagtc
     4681 tcatccccct gtggtaccat cttgtcctca gaggtggttt gtgctccccc tactgcctat
     4741 atcgacttcg cccggcagaa gctagatccc aagattgctg tggctgcgca gaactgctac
     4801 aaagtgacta atggggcttt tactggggag atcaggtgag atcgaggtgg agaggggtgt
     4861 gtgggaccct tccctcactt tcctcgttga ggggaaagcc acagggtggg ctccctgctg
     4921 aaccttggct tcatctcttc ctttagccct ggcatgatca aagactgcgg agccacgtgg
     4981 gtggtcctgg ggcactcaga gagaaggcat gtctttgggg agtcagatga ggttagtagc
     5041 caagagagaa gataagggat gtctttttcc aagaaggatg tctcaccaag tctgtttctc
     5101 aacagctgat tgggcagaaa gtggcccatg ctctggcaga gggactcgga gtaatcgcct
     5161 gcattgggga gaagctagat gaaagggaag ctggcatcac tgagaaggtt gttttcgagc
     5221 agacaaaggt catcgcaggt atctctggag aaagggacct ttgagcctat ccagggccac
     5281 agagactcag agggtagggt caggccctgg agcctgtctt ggtccccatg ctgatccaga
     5341 aaaggaaaaa ggggaggggg agtgacaatc tttgcttggg gcctatgact tctccagccc
     5401 caaggtagat gccacctgga aatcccccaa tgtccactag ggggcagtag gccaccgttc
     5461 ttcgtactcc ggagaacctg gctggagagc tctttcttgt tcacccttcc ctccatctgt
     5521 atctctgccc tgcagataac gtgaaggact ggagcaaggt cgtcctggcc tatgagcctg
     5581 tgtgggccat tggtactggc aagactgcaa caccccaaca ggtaaccggg cccaggagcc
     5641 ctgccctcat cccagcctgc ctcaataggt ttggacagac acagcccaca tggggcaacc
     5701 ccttatttca aagacacaga gaccttgaac ccagagacag tgacttgtcc aagggcatcc
     5761 agtccagggc ctggcttgga tcagagccct ggtactctga ctcagtcaga aaccacacta
     5821 agtgtccact ggtgccagtg atttttcctc ttagagaggc agaaaaggtc ttacttaggc
     5881 cagcttcttg ttctaggccc aggaagtaca cgagaagctc cgaggatggc tgaagtccaa
     5941 cgtctctgat gcggtggctc agagcacccg tatcatttat ggaggtgagt ggctttggtt
     6001 cccggctgag gtggagtggg ctgaggacta gactgagccc tcggacatgg aggtggggat
     6061 ggggcagact catcccattc ttgaccaagc ccttgttctg ctcccttccc aggctctgtg
     6121 actggggcaa cctgcaagga gctggccagc cagcctgatg tggatggctt ccttgtgggt
     6181 ggtgcttccc tcaagcccga attcgtggac atcatcaatg ccaaacaatg agccccatcc
     6241 atcttcccta cccttcctgc caagccaggg actgctagcc cggttgtaca ggtaccggat
     6301 cctaaggtac ctaattgcct agaaaggagc agacgatatg gcgtcgctcc ctgcaggtcg
     6361 actctagaaa ccagcagagc atatgggctc gctggctgca gtattcccgg gttcattaga
     6421 tcctaaggta cctaattgcc tagaaaggag cagacgatat ggcgtcgctc cctgcaggtc
     6481 gactctagaa accagcagag catatgggct cgctggctgc agtattcccg ggttcattag
     6541 atcctaaggt acctaattgc ctagaaagga gcagacgata tggcgtcgct ccctgcaggt
     6601 cgactctaga aaccagcaga gcatatgggc tcgctggctg cagtattccc gggttcatta
     6661 gatcctaagg tacctaattg cctagaaagg agcagacgat atggcgtcgc tccctgcagg
     6721 tcgactctag aaaccagcag agcatatggg ctcgctggct gcagtattcc cgggttcatt
     6781 agatcctaag gtacctaatt gcctagaaag gagcagacga tatggcgtcg ctccctgcag
     6841 gtcgactcta gaaaccagca gagcatatgg gctcgctggc tgcagtattc ccgggttcat
     6901 tagatcctaa ggtacctaat tgcctagaaa ggagcagacg atatggcgtc gctccctgca
     6961 ggtcgactct agaaaccagc agagcatatg ggctcgctgg ctgcagtatt cccgggttca
     7021 ttagatccta aggtacctaa ttgcctagaa aggagcagac gatatggcgt cgctccctgc
     7081 aggtcgactc tagaaaccag cagagcatat gggctcgctg gctgcagtat tcccgggttc
     7141 attagatcct aaggtaccta attgcctaga aaggagcaga cgatatggcg tcgctccctg
     7201 caggtcgact ctagaaacca gcagagcata tgggctcgct ggctgcagta ttcccgggtt
     7261 cattagatcc taaggtacct aattgcctag aaaggagcag acgatatggc gtcgctccct
     7321 gcaggtcgac tctagaaacc agcagagcat atgggctcgc tggctgcagt attcccgggt
     7381 tcattagatc ctaaggtacc taattgccta gaaaggagca gacgatatgg cgtcgctccc
     7441 tgcaggtcga ctctagaaac cagcagagca tatgggctcg ctggctgcag tattcccggg
     7501 ttcattagat cctaaggtac ctaattgcct agaaaggagc agacgatatg gcgtcgctcc
     7561 ctgcaggtcg actctagaaa ccagcagagc atatgggctc gctggctgca gtattcccgg
     7621 gttcattaga tcctaaggta cctaattgcc tagaaaggag cagacgatat ggcgtcgctc
     7681 cctgcaggtc gactctagaa accagcagag catatgggct cgctggctgc agtattcccg
     7741 ggttcattag atctcgcgaa gggcgaattc acgaggcgcg cccggatcgc gatcgcaatc
     7801 ttaattaact cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     7861 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     7921 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     7981 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     8041 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     8101 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     8161 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     8221 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     8281 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     8341 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     8401 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     8461 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     8521 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     8581 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     8641 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     8701 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     8761 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     8821 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     8881 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt
     8941 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
     9001 taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga
     9061 gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc
     9121 gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg
     9181 ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg
     9241 tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc
     9301 gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc
     9361 gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat
     9421 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     9481 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     9541 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     9601 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     9661 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     9721 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     9781 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     9841 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     9901 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     9961 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
    10021 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
    10081 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
    10141 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
    10201 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
    10261 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
    10321 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
    10381 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
    10441 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
    10501 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
    10561 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
    10621 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
    10681 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
    10741 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
    10801 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
    10861 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
    10921 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
    10981 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
    11041 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
    11101 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
    11161 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
    11221 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
    11281 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
    11341 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
    11401 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
    11461 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
    11521 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
    11581 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
    11641 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
    11701 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
    11761 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
    11821 aaagtgccac ctgacgtc
//
