LOCUS       sequence-415818-       10268 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Protein quality control substrate and associated control YIp 
            ADE2-LEU2 pTDH3::lys1-W151D-GFP::tADH1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10268)
  AUTHORS   Flagg MP, Lam B, Lam DK, Le TM, Kao A, Slaiwa YI, Hampton RY
  TITLE     Exploring the ""misfolding problem"" by systematic discovery and 
            analysis of functional-but-degraded proteins.
  JOURNAL   Mol Biol Cell. 2023 Dec 1;34(13):ar125. doi: 
            10.1091/mbc.E23-06-0248. Epub 2023 Sep 20.
  PUBMED    37729018
REFERENCE   2  (bases 1 to 10268)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Biol 
            Cell."; date: "2023-12-1"; volume: "34(13):ar125. doi"; pages: " 
            10.1091/mbc.E23-06-0248. Epub 2023 Sep 20"
COMMENT     Sequence Label: pRH3214
FEATURES             Location/Qualifiers
     source          1..10268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..665
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     558..581
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     complement(697..713)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(698..714)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             1863..2573
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1890..1918)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2513..2533
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2597..2784
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2652..2676)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2788..2807)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2789..2807)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2817..2834)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2817..2833)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2826..2848)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2974..3429
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3061..3080)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3271..3292
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3634..5349)
                     /codon_start=1
                     /gene="S. cerevisiae ADE2"
                     /product="phosphoribosylaminoimidazole carboxylase, 
                     required for adenine biosynthesis"
                     /label=ADE2
                     /note="yeast auxotrophic marker"
                     /translation="MDSRTVGILGGGQLGRMIVEAANRLNIKTVILDAENSPAKQISNS
                     NDHVNGSFSNPLDIEKLAEKCDVLTIEIEHVDVPTLKNLQVKHPKLKIYPSPETIRLIQ
                     DKYIQKEHLIKNGIAVTQSVPVEQASETSLLNVGRDLGFPFVLKSRTLAYDGRGNFVVK
                     NKEMIPEALEVLKDRPLYAEKWAPFTKELAVMIVRSVNGLVFSYPIVETIHKDNICDLC
                     YAPARVPDSVQLKAKLLAENAIKSFPGCGIFGVEMFYLETGELLINEIAPRPHNSGHYT
                     IDACVTSQFEAHLRSILDLPMPKNFTSFSTITTNAIMLNVLGDKHTKDKELETCERALA
                     TPGSSVYLYGKESRPNRKVGHINIIASSMAECEQRLNYITGRTDIPIKISVAQKLDLEA
                     MVKPLVGIIMGSDSDLPVMSAACAVLKDFGVPFEVTIVSAHRTPHRMSAYAISASKRGI
                     KTIIAGAGGAAHLPGMVAAMTPLPVIGVPVKGSCLDGVDSLHSIVQMPRGVPVATVAIN
                     NSTNAALLAVRLLGAYDSSYTTKMEQFLLKQEEEVLVKAQKLETVGYEAYLENK"
     promoter        complement(5350..5725)
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     primer_bind     complement(5916..5935)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6035..6057
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6095..6113)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(6399..7493)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(7494..7901)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     promoter        8069..8173
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8174..8242,8243..9034)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8392..8411)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9205..9793
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9694..9713
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9947..9964
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10081..10102
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10117..10122,10123..10140,10141..10147)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10155..10171
                     /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)."
     primer_bind     10160..10182
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10179..10195
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10179..10195
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10214..10234
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10216..10234
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gctgcttggc ggtcgagttt atcattatca atactgccat ttcaaagaat acgtaaataa
       61 ttaatagtag tgattttcct aactttattt agtcaaaaaa ttagcctttt aattctgctg
      121 taacccgtac atgcccaaaa tagggggcgg gttacacaga atatataaca tcgtaggtgt
      181 ctgggtgaac agtttattcc tggcatccac taaatataat ggagcccgct ttttaagctg
      241 gcatccagaa aaaaaaagaa tcccagcacc aaaatattgt tttcttcacc aaccatcagt
      301 tcataggtcc attctcttag cgcaactaca gagaacaggg gcacaaacag gcaaaaaacg
      361 ggcacaacct caatggagtg atgcaacctg cctggagtaa atgatgacac aaggcaattg
      421 acccacgcat gtatctatct cattttctta caccttctat taccttctgc tctctctgat
      481 ttggaaaaag ctgaaaaaaa aggttgaaac cagttccctg aaattattcc cctacttgac
      541 taataagtat ataaagacgg taggtattga ttgtaattct gtaaatctat ttcttaaact
      601 tcttaaattc tacttttata gttagtcttt tttttagttt taaaacacca agaacttagt
      661 ttcgacggat ctgcaggaat tcgatatcaa gcttatcgat accgtcgacc tcgagaatgg
      721 ctgccgtcac attacatcta agagctgaaa ctaaacccct agaggcacgt gctgccttaa
      781 cacctaccac ggttaaaaaa ctgatagcta agggcttcaa aatatatgta gaggacagtc
      841 cacaatctac tttcaatatt aacgaatatc gtcaagcagg tgccattata gtgcctgcag
      901 gttcatggaa aaccgctcca cgcgacagaa tcattatagg tttgaaggaa atgcctgaaa
      961 ccgatacttt ccctctagtc cacgaacaca tccagtttgc tcactgctac aaagaccaag
     1021 ctgggtggca aaatgtcctt atgagattta ttaagggaca cggtactcta tatgatttgg
     1081 aatttttgga aaatgaccaa ggtagaagag ttgctgcctt tggattttac gctgggttcg
     1141 caggtgcagc ccttggtgta agagacgatg cattcaagca aacgcattct gacgatgaag
     1201 acttgcctgc agtgtcgcct taccccaatg aaaaggcatt ggttaaagat gttaccaaag
     1261 attataaaga agccttagcc accggagcca gaaagccaac cgtgttaatc attggtgcgc
     1321 taggaagatg tggttccggt gccatcgatc tgttgcacaa agttggtatt ccagatgcta
     1381 acatattaaa atgggatatc aaagaaactt cccgtggtgg tccctttgac gaaattccac
     1441 aagctgatat ttttatcaat tgtatatatc tatcgaagcc aattgctcct ttcactaaca
     1501 tggagaaact gaataatcct aacagaagac taaggaccgt ggtggacgta tcagcagaca
     1561 ctaccaaccc tcacaacccc atcccaatat acactgtggc tactgtgttt aacaaaccta
     1621 ccgttctggt acctaccact gccgggccta aattatctgt catctctatt gatcacttgc
     1681 cttctttgct gccaagagaa gcttcagaat ttttctctca tgatctctta ccatctttag
     1741 agctcctacc tcaaagaaaa actgctcctg tctgggttag agccaagaaa ttgttcgata
     1801 gacattgcgc tcgtgttaaa agatcttcaa gattgggtcg acggatcccc gggttaatta
     1861 acagtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg
     1921 atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa
     1981 aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg ccaacacttg
     2041 tcactacttt cacttatggt gttcaatgct tttcaagata cccagatcat atgaaacggc
     2101 atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca
     2161 aagatgacgg gaactacaag acacgtgctg aagtcaagtt tgaaggtgat acccttgtta
     2221 atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt ggacacaaat
     2281 tggaatacaa ctataactca cacaatgtat acatcatggc agacaaacaa aagaatggaa
     2341 tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa ctagcagacc
     2401 attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac aaccattacc
     2461 tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa gagagaccac atggtccttc
     2521 ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac aaatagggcg
     2581 cgccacttct aaataagcga atttcttatg atttatgatt tttattatta aataagttat
     2641 aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt
     2701 attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagt atgaggtcgc
     2761 tcttattgac cacacctcta ccggcagccc tatagtgagt cgtattacgc gcgctcactg
     2821 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     2881 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     2941 tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg
     3001 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3061 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3121 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3181 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3241 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3301 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3361 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     3421 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3481 agatcttatg tatgaaattc ttaaaaaagg acacctgtaa gcgttgattt ctatgtatga
     3541 agtccacatt tgatgtaatc ataacaaagc ctaaaaaata ggtatatcat tttataatta
     3601 tttgctgtac aagtatatca ataaacttat atattacttg ttttctagat aagcttcgta
     3661 accgacagtt tctaactttt gtgctttgac aagaacttct tcttcttgct ttaataaaaa
     3721 ctgttccatt ttcgttgtat aacttgaatc ataagcgcca agcagtctga cagccaacag
     3781 cgcagcgttc gtactattat taatagcgac ggtagctact ggaacacctc taggcatttg
     3841 cacaattgaa tgtaaagaat ctactccatc tagacaagaa ccttttacgg gcacaccgat
     3901 gacaggaagt ggtgtcattg cagccaccat acctggcaag tgagcagccc caccagctcc
     3961 agcgataatt gttttaattc cacgcttgct tgcggaaata gcatatgctg acatcctatg
     4021 tggagttcta tgagcagaga ctattgtcac ttcaaatgga acgccaaaat cttttaaaac
     4081 cgcacatgcg gcagacatta ccggcaagtc agagtctgat cccatgatga ttccaaccaa
     4141 tggtttgacc attgcttcca agtccaactt ttgagcgaca gagattttga ttggaatatc
     4201 agttctacct gtaatgtagt tcagcctttg ttcacattcc gccatactgg aggcaataat
     4261 atttatgtga cctacttttc tgttaggtct agactctttt ccatataagt acactgagga
     4321 acctggagtc gccaatgctc tttcgcaagt ttctagctct ttatcttttg tatgtttgtc
     4381 tccaagaaca tttagcataa tggcgttcgt tgtaatggtg gagaaagatg tgaaattctt
     4441 tggcattggc aaatccaata ttgatctcaa atgagcttca aattgagaag tgacgcaagc
     4501 atcaatggta taatgtccag agttgtgagg ccttggggca atttcgttaa taagcaattc
     4561 ccctgtttct aaatagaaca tttccacacc aaatatacca caaccgggaa aagatttgat
     4621 tgcattttct gccaacaact tcgccttaag ttgaacggag tccggaactc tagcaggcgc
     4681 ataacataag tcacaaatat tgtccttgtg gatagtctct acaattgggt aagaaaacac
     4741 taaaccgtta acagacctca caatcatgac tgctaattct ttagtaaatg gtgcccattt
     4801 ttcggcgtac aaaggacgat ccttcagtac ttccaaagct tccggaatca tttccttatt
     4861 ctttacaacg aagttacctc ttccatcgta tgccaaagtc ctcgacttca agacgaatgg
     4921 aaaacccaaa tctcttccaa cattcaatag ggacgtctca ctggcttgtt ccacaggaac
     4981 actttgggta actgctatac catttttgat taaatgctct ttttgaatat atttgtcttg
     5041 tatcaatctg attgtttctg gagaagggta aatttttaat ttgggatgtt ttacttgaag
     5101 attctttagt gtaggaacat caacatgctc aatctcaatc gttagcacat cacatttttc
     5161 agctagtttt tcgatatcaa gaggattgga aaaggagcca ttaacgtggt cattggagtt
     5221 gcttatttgt ttggcaggag aattttcagc atctagtatt accgtcttaa tgttgagcct
     5281 gtttgctgcc tcaacaatca tacgtcccaa ttgtccccct cctaatatac caactgttct
     5341 agaatccata cttgattgtt ttgtccgatt ttcttgtttt tcttgattgt tatagtagga
     5401 tgtacttaga agagagatcc aacgatttta cgcaccaatt tatacatgaa atgctccata
     5461 atattgtcca tttagttctt aataaaaggt cagcaagagt caatcactta gtattacccg
     5521 gttcgtagcc atgcaacaag agtcatttgt cagcatagct gtaataatca atcatgacgt
     5581 aagaaatgta tcataattaa aagttgttaa agatgtcagt gttatgttgg tgttacaaaa
     5641 ttctcggctt ctcactaata tttaatatct cttaaatttt atctgtcttt gattctttta
     5701 agaaaagtta tgtattattc aagaattaat tcagatctat tcttttgatt tataagggat
     5761 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5821 ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
     5881 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag
     5941 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     6001 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     6061 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     6121 gttaatgtca tgatcttttt gtgtggtgcc ctcctccttg tcaatattaa tgttaaagtg
     6181 caattctttt tccttatcac gttgagccat tagtatcaat ttgcttacct gtattccttt
     6241 acatcctcct ttttctcctt cttgataaat gtatgtagat tgcgtatata gtttcgtcta
     6301 ccctatgaac atattccatt ttgtaatttc gtgtcgtttc tattatgaat ttcatttata
     6361 aagtttatgt acaaatatca taaaaaaaga gaatcttttt aagcaaggat tttcttaact
     6421 tcttcggcga cagcatcacc gacttcggtg gtactgttgg aaccacctaa atcaccagtt
     6481 ctgatacctg catccaaaac ctttttaact gcatcttcaa tggccttacc ttcttcaggc
     6541 aagttcaatg acaatttcaa catcattgca gcagacaaga tagtggcgat agggttgacc
     6601 ttattctttg gcaaatctgg agcagaaccg tggcatggtt cgtacaaacc aaatgcggtg
     6661 ttcttgtctg gcaaagaggc caaggacgca gatggcaaca aacccaagga acctgggata
     6721 acggaggctt catcggagat gatatcacca aacatgttgc tggtgattat aataccattt
     6781 aggtgggttg ggttcttaac taggatcatg gcggcagaat caatcaattg atgttgaacc
     6841 ttcaatgtag ggaattcgtt cttgatggtt tcctccacag tttttctcca taatcttgaa
     6901 gaggccaaaa cattagcttt atccaaggac caaataggca atggtggctc atgttgtagg
     6961 gccatgaaag cggccattct tgtgattctt tgcacttctg gaacggtgta ttgttcacta
     7021 tcccaagcga caccatcacc atcgtcttcc tttctcttac caaagtaaat acctcccact
     7081 aattctctga caacaacgaa gtcagtacct ttagcaaatt gtggcttgat tggagataag
     7141 tctaaaagag agtcggatgc aaagttacat ggtcttaagt tggcgtacaa ttgaagttct
     7201 ttacggattt ttagtaaacc ttgttcaggt ctaacactac cggtacccca tttaggacca
     7261 cccacagcac ctaacaaaac ggcatcagcc ttcttggagg cttccagcgc ctcatctgga
     7321 agtggaacac ctgtagcatc gatagcagca ccaccaatta aatgattttc gaaatcgaac
     7381 ttgacattgg aacgaacatc agaaatagct ttaagaacct taatggcttc ggctgtgatt
     7441 tcttgaccaa cgtggtcacc tggcaaaacg acgatcttct taggggcaga cattagaatg
     7501 gtatatcctt gaaatatata tatatatatt gctgaaatgt aaaaggtaag aaaagttaga
     7561 aagtaagacg attgctaacc acctattgga aaaaacaata ggtccttaaa taatattgtc
     7621 aacttcaagt attgtgatgc aagcatttag tcatgaacgc ttctctattc tatatgaaaa
     7681 gccggttccg gcgctctcac ctttcctttt tctcccaatt tttcagttga aaaaggtata
     7741 tgcgtcaggc gacctctgaa attaacaaaa aatttccagt catcgaattt gattctgtgc
     7801 gatagcgccc ctgtgtgttc tcgttatgtt gaggaaaaaa ataatggttg ctaagagatt
     7861 cgaactcttg catcttacga tacctgagta ttcccacagt taactgcggt caagatattt
     7921 cttgaatcag gcgccttaga ccgctcggcc aaacaaccaa ttacttgttg agaaatagag
     7981 tataattatc ctataaatat aacgtttttg aacacacatg aacaaggaag tacaggagac
     8041 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     8101 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     8161 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     8221 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     8281 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     8341 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     8401 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     8461 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     8521 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     8581 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     8641 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     8701 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     8761 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     8821 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     8881 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     8941 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     9001 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     9061 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     9121 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     9181 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     9241 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     9301 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     9361 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     9421 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     9481 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     9541 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     9601 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     9661 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     9721 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     9781 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     9841 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     9901 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     9961 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
    10021 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
    10081 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
    10141 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
    10201 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc tccaccgcgg
    10261 tggcggcc
//
