LOCUS       sequence-476828-       10240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the reverse tetracycline activator (rtTA) and T7 
            polymerase; Plasmid #1 of the tet-on overexpression system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10240)
  AUTHORS   Nostramo RT, Sinopoli PL, Bao A, Metcalf S, Peltier LM, Hopper AK
  TITLE     Free introns of tRNAs as complementarity-dependent regulators of 
            gene expression.
  JOURNAL   Mol Cell. 2025 Feb 20;85(4):726-741.e6. doi: 
            10.1016/j.molcel.2025.01.019. Epub 2025 Feb 11.
  PUBMED    39938518
REFERENCE   2  (bases 1 to 10240)
  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.2025.01.019"; journalName: "Mol Cell"; date: 
            "2025-02-20- 20"; volume: "85"; issue: "4"; pages: "726-741.e6"
COMMENT     Sequence Label: TDH3pro-rtTA-tetO7pro-T7pol-pRS413
FEATURES             Location/Qualifiers
     source          1..10240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        317..504
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             505..1167
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(1427..1882)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1564..1585)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1776..1795
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2008..2030
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2022..2039
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2023..2039
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2049..2068
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2049..2067
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2114..2303)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2315..2333
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(4968..4988)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    join(5224..5242,5243..5265,5266..5284,5285..5307,5308..5326,
                     5327..5350,5351..5369,5370..5392,5393..5411,5412..5434,
                     5435..5453,5454..5476,5477..5495)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     CDS             complement(join(5742..6125,6126..6746))
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /label=tTA
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKIINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEPESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTI
                     EGLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHA
                     DALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     6654..6673
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        complement(6747..7414)
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     complement(6854..6877)
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     complement(7467..7487)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7467..7485)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7506..7522)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7506..7522)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7519..7541)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7530..7546
                     /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(7554..7560,7561..7578,7579..7584))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7599..7620
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7737..7754)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7908..8496)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7988..8007)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8667..9458,9459..9527))
                     /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     9290..9309
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9528..9632)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    9662..10165
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     10213..10231
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaatt cccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt
      241 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta
      301 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat
      361 tttttttttt ccacctagcg gatgactctt tttttttctt agcgattggc attatcacat
      421 aatgaattat acattatata aagtaatgtg atttcttcga agaatatact aaaaaatgag
      481 caggcaagat aaacgaaggc aaagatgaca gagcagaaag ccctagtaaa gcgtattaca
      541 aatgaaacca agattcagat tgcgatctct ttaaagggtg gtcccctagc gatagagcac
      601 tcgatcttcc cagaaaaaga ggcagaagca gtagcagaac aggccacaca atcgcaagtg
      661 attaacgtcc acacaggtat agggtttctg gaccatatga tacatgctct ggccaagcat
      721 tccggctggt cgctaatcgt tgagtgcatt ggtgacttac acatagacga ccatcacacc
      781 actgaagact gcgggattgc tctcggtcaa gcttttaaag aggccctagg ggccgtgcgt
      841 ggagtaaaaa ggtttggatc aggatttgcg cctttggatg aggcactttc cagagcggtg
      901 gtagatcttt cgaacaggcc gtacgcagtt gtcgaacttg gtttgcaaag ggagaaagta
      961 ggagatctct cttgcgagat gatcccgcat tttcttgaaa gctttgcaga ggctagcaga
     1021 attaccctcc acgttgattg tctgcgaggc aagaatgatc atcaccgtag tgagagtgcg
     1081 ttcaaggctc ttgcggttgc cataagagaa gccacctcgc ccaatggtac caacgatgtt
     1141 ccctccacca aaggtgttct tatgtagtga caccgattat ttaaagctgc agcatacgat
     1201 atatatacat gtgtatatat gtatacctat gaatgtcagt aagtatgtat acgaacagta
     1261 tgatactgaa gatgacaagg taatgcatca ttctatacgt gtcattctga acgaggcgcg
     1321 ctttcctttt ttctttttgc tttttctttt tttttctctt gaactcgacg gatctatgcg
     1381 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt
     1441 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     1501 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     1561 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     1621 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     1681 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     1741 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     1801 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     1861 aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc
     1921 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     1981 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     2041 agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc ctcgaggtcg
     2101 accaaattaa agccttcgag cgtcccaaaa ccttctcaag caaggttttc agtataatgt
     2161 tacatgcgta cacgcgtctg tacagaaaaa aaagaaaaat ttgaaatata aataacgttc
     2221 ttaatactaa cataactata aaaaaataaa tagggaccta gacttcaggt tgtctaactc
     2281 cttccttttc ggttagagcg gatgtggggg gagggcgtga atgtaagcgt gacataacta
     2341 atttacgcga acgcgaagtc cgactctaag atgtcacgga ggttcaagtt acctttagcc
     2401 ggaagtgctg gcattttgtc caattgagac tcgtgcaact ggtcagcgaa ctggtcgtag
     2461 aaatcagcca gtacatcaca agactcatat gtgtcaacca tagtttcgcg cactgctttg
     2521 aacaggttcg cagcgtcagc cggaatggta ccgaaggagt cgtgaatcag tgcaaaagat
     2581 tcgattccgt acttctcgtg tgcccacact acagtcttac gaaggtggct accgtcttgg
     2641 ctgtgtacaa agttaggagc gataccagac tcctgtttgt gtgcatcaat ctcgctatct
     2701 ttgttggtgt taatggtagg ctgtaagcgg aactgaccga ggaacatcag gttcaagcgc
     2761 gtctgaatag gcttcttgta ttcctgccac acagggaaac catcaggagt tacccaatgc
     2821 acagcgcaac gcttgcgaag aatctctcca gtcttcttat ctttgacctc agcagccagc
     2881 agcttagcag cagacttaag ccagttcatt gcttcaaccg cagctaccac cgtcacgctc
     2941 acagattccc aaatcagctt agccatgtat ccagcagcct gattcggctg agtgaacatc
     3001 agacccttgc cggaatcaat agctggctga atggtatctt ccagcacttg ttgacggaag
     3061 ccgaactctt tggacccgta agccagcgtc atgactgaac gcttagtcac actgcgagta
     3121 acaccgtaag ccagccattg accagccagt gccttagtgc ccagcttgac tttctcagag
     3181 atttcaccag tgttctcatc ggtcacggta actacttcgt tatcggtccc attgattgcg
     3241 tctgcttgta gaatctcgtt gactttctta gcaacaatcc cgtagatgtc ctgaacggtt
     3301 tcactaggaa gcaagttaac cgcgcgacca cctacctcat ctcggagcat cgcggagaag
     3361 tgctggatgc cagagcaaga cccgtcaaac gccagcggaa gggagcagtt atagctcagg
     3421 ccgtggtgct gtaccccagc gtactcaaag cagaacgcaa ggaagcagaa cggagaatct
     3481 tgctcagccc accaagtgtt ctccagtgga gacttagcgc aagccatgat gttctcgtgg
     3541 ttttcctcaa tgaacttgat gcgctcaggg aacggaacct tatcgacacc cgcacagttt
     3601 gcaccgtgga ttttcagcca gtagtaacct tccttaccga ttggtttacc tttcgccagc
     3661 gtaagcagtc ctttggtcat atcgttacct tgcgggttga acattgacac agcgtaaaca
     3721 cgaccgcgcc agtccatgtt gtaagggaac cagatggcct tatggttagc aaacttattg
     3781 gcttgctcaa gcatgaactc aaggctgata cggcgagact tgcgagcctt gtccttgcgg
     3841 tacacagcag cggcagcacg tttccacgcg gtgagagcct caggattcat gtcgatgtct
     3901 tccggtttca tcgggagttc ttcacgctca atcgcaggga tgtcctcgac cggacaatgc
     3961 ttccacttgg tgattacgtt ggcgaccgct aggactttct tgttgatttt ccatgcggtg
     4021 ttttgcgcaa tgttaatcgc tttgtacacc tcaggcatgt aaacgtcttc gtagcgcatc
     4081 agtgctttct tactgtgagt acgcaccagc gccagaggac gacgaccgtt agcccaatag
     4141 ccaccaccag taatgccagt ccacggctta ggaggaacta cgcaaggttg gaacatcaga
     4201 gagatgccag ccagcgcacc tgcacgggtt gcgatagcct cagcgtattc aggtgcgagt
     4261 tcgatagtct cagagtcttg acctactacg ccagcatttt ggcggtgtaa gctaaccatt
     4321 ccggttgact caatgagcat ctcgatgcag cgtactccta catgaataga gtcttcctta
     4381 tgccacgaag accacgcctc gccaccgagt agacccttag agagcatgtc agcctcgaca
     4441 acttgcataa atgctttctt gtagacgtgc cctacgcgct tgttgagttg ttcctcaacg
     4501 tttttcttga agtgcttagc ttcaaggtca cggatacgac cgaagcgagc ctcgtcctca
     4561 atggcccgac cgattgcgct tgctacagcc tgaacggttg tattgtcagc actggttagg
     4621 caagccagag tggtcttaat ggtgatgtac gctacggctt ccggcttgat ttcttgcagg
     4681 aactggaagg ctgtcgggcg cttgccgcgc ttagctttca cttcctcaaa ccagtcgttg
     4741 atgcgtgcaa tcatcttagg gagtagggta gtgatgagag gcttggcggc agcgttatcc
     4801 gcaacctcac cagctttaag ttgacgctca aacatcttgc ggaagcgtgc ttcacccatc
     4861 tcgtaagact catgctcaag ggccaactgt tcgcgagcta aacgctcacc gtaatggtca
     4921 gccagagtgt tgaacgggat agcagccagt tcgatgtcag agaattcgac ctttctcttc
     4981 ttttttggag gctcgaggaa ttcgttctta gcgatgttaa tcgtgttcat ggatccgacc
     5041 ccccgatgat ccggtaattt agtgtgtgta tttgtgtttg cgtgtctata gaagtatagt
     5101 aatttatgct acaaaggacc taatgtataa ggaaagaata tttagagaaa agaagaaaac
     5161 aagagtttta tatacataca gagcacatgc atgccatagg gtaccgagct cgactttcac
     5221 ttttctctat caccgatagg gagtggtaaa ctcgactttc acttttctct atcactgata
     5281 gggagtggta aactcgactt tcacttttct ctatcactga tagggagtgg taaacctcga
     5341 ctttcacttt tctctatcac cgatagggag tggtaaactc gactttcact tttctctatc
     5401 actgataggg agtggtaaac tcgactttca cttttctcta tcactgatag ggagtggtaa
     5461 actcgacttt cacttttctc tatcactgat agggagtggt aaactcgatc gaggaattga
     5521 tctatattac ccgcggccgc ctaaaggcat taaaagagga gcgaattttt ttttaataaa
     5581 aatcttaata atcattaaaa gataaataat agtctatata tacgtatata aataaaaaat
     5641 attcaaaaaa taaaataaac tattatttta gcgtaaagga tggggaaaga gaaaagaaaa
     5701 aaattgatct atcgatttca attcaattca atttatctag tcccaccgta ctcgtcaatt
     5761 ccaagggcat cggtaaacat ctgctcaaac tcgaagtcgg ccatatccag agcgccgtag
     5821 ggggcggagt cgtggggggt aaatcccgga cccggggaat ccccgtcccc caacatgtcc
     5881 agatcgaaat cgtctagcgc gtcggcatgc gccatcgcca cgtcctcgcc gtctaagtgg
     5941 agctcgtccc ccaggctgac atcggtcggg ggggccgtcg acagtctgcg cgtgtgtccc
     6001 gcggggagaa aggacaggcg cggagccgcc agccccgcct cttcgggggc gtcgtcgtcc
     6061 gggagatcga gcaggccctc gatggtagac ccgtaattgt ttttcgtacg cgcgcggctg
     6121 tacgcggacc cactttcaca tttaagttgt ttttctaatc cgcatatgat caattcaagg
     6181 ccgaataaga aggctggctc tgcaccttgg cgatcaaata attcgatagc ttgttttaat
     6241 aatggcggca tactatcagt agtaggtgtt tccctttctt ctttagcgac ttgatgctct
     6301 tgttcttcca atacgcaacc taaagtaaag tgccccacag cgctgagtgc atataatgca
     6361 ttctctagtg aaaaaccttg ttggcataaa aaggctaatt gattttcgag agtttcatac
     6421 tgtttttctg taggccgtgt acctaaatgt acttttgctc catcgcgatg acttagtaaa
     6481 gcacatctat aacttttagc gttattacgt aaaaaatctt gccagctttc aggttctaaa
     6541 gggcaagagt gagtatggtg cctatctaac atctcaatgg gtaaggcgtc gagcaaagcc
     6601 cgcttatttt ttacatgcca atacaatgta ggctgctcta cacctagctt ctgggcgagt
     6661 ttacgggttg ttaaaccttc gattccgacc ccattaagca gctctaatgc gccgttaata
     6721 attttacttt tatctaatct agacattttg tttgtttatg tgtgtttatt cgaaactaag
     6781 ttcttggtgt tttaaaacta aaaaaaagac taactataaa agtagaattt aagaagttta
     6841 agaaatagat ttacagaatt acaatcaata cctaccgtct ttatatactt attagtcaag
     6901 taggggaata atttcaggga actggtttca accttttttt tcagcttttt ccaaatcaga
     6961 gagagcagaa ggtaatagaa ggtgtaagaa aatgagatag atacatgcgt gggtcaattg
     7021 ccttgtgtca tcatttactc caggcaggtt gcatcactcc attgaggttg tgcccgtttt
     7081 ttgcctgttt gtgcccctgt tctctgtagt tgcgctaaga gaatggacct atgaactgat
     7141 ggttggtgaa gaaaacaata ttttggtgct gggattcttt tttttttctg gatgccagct
     7201 taaaaagcgg gctccattat atttagtgga tgccaggaat aaactgttca cccagacacc
     7261 tacgatgtta tatattctgt gtaacccgcc ccctattttg ggcatgtacg ggttacagca
     7321 gaattaaaag gctaattttt tgactaaata aagttaggaa aatcactact attaattatt
     7381 tacgtattct ttgaaatggc agtattgata atgaaagctt atcgataccg gcggccgcca
     7441 ccgcggtgga gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     7501 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     7561 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     7621 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     7681 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     7741 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     7801 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     7861 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     7921 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     7981 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     8041 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     8101 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     8161 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     8221 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     8281 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     8341 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     8401 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     8461 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     8521 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     8581 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     8641 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     8701 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     8761 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     8821 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     8881 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     8941 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     9001 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     9061 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     9121 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     9181 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     9241 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     9301 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     9361 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     9421 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     9481 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     9541 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     9601 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     9661 ggatcgcttg cctgtaactt acacgcgcct cgtatctttt aatgatggaa taatttggga
     9721 atttactctg tgtttattta tttttatgtt ttgtatttgg attttagaaa gtaaataaag
     9781 aaggtagaag agttacggaa tgaagaaaaa aaaataaaca aaggtttaaa aaatttcaac
     9841 aaaaagcgta ctttacatat atatttatta gacaagaaaa gcagattaaa tagatataca
     9901 ttcgattaac gataagtaaa atgtaaaatc acaggatttt cgtgtgtggt cttctacaca
     9961 gacaagatga aacaattcgg cattaatacc tgagagcagg aagagcaaga taaaaggtag
    10021 tatttgttgg cgatccccct agagtctttt acatcttcgg aaaacaaaaa ctattttttc
    10081 tttaatttct ttttttactt tctattttta atttatatat ttatattaaa aaatttaaat
    10141 tataattatt tttatagcac gtgatgaaaa ggaccctaag aaaccattat tatcatgaca
    10201 ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
