LOCUS       sequence-422772-       11038 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-Off controlled mRuby3-Pho8 with TRP1 selection. 
            Tetracycline-controlled transactivator (tTA) present on same 
            plasmid. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11038)
  AUTHORS   He CW, Cui XF, Ma SJ, Xu Q, Ran YP, Chen WZ, Mu JX, Li H, Zhu J, 
            Gong Q, Xie Z
  TITLE     Membrane recruitment of Atg8 by Hfl1 facilitates turnover of 
            vacuolar membrane proteins in yeast cells approaching stationary 
            phase.
  JOURNAL   BMC Biol. 2021 Jun 4;19(1):117. doi: 10.1186/s12915-021-01048-7.
  PUBMED    34088313
REFERENCE   2  (bases 1 to 11038)
  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: "BMC Biol.";
            date: "2021-06-4"; volume: "19(1):117. doi"; pages: " 
            10.1186/s12915-021-01048-7"
COMMENT     Sequence Label: BS-TRP1-TetOff(7 repeats)-mRuby3-Pho8
FEATURES             Location/Qualifiers
     source          1..11038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..21
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5..21
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     40..60
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        42..60
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        796..1073
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     1810..1826
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1811..1827
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(join(1863..2249,2250..2870))
                     /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="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTI
                     EGLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHA
                     DALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     2778..2797
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(2955..2979)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2976..3179)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3005..3025)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3180..3559
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     terminator      3671..3858
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3726..3750)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     protein_bind    join(3916..3934,3935..3957,3958..3976,3977..3999,4000..4018,
                     4019..4041,4042..4060,4061..4083,4084..4102,4103..4125,
                     4126..4144,4145..4167,4168..4186)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     CDS             4410..5120
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     primer_bind     complement(7225..7241)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7225..7241)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(8268..8287)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8269..8287)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8294..8311)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8294..8310)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8303..8325)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8452..8907
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8539..8558)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8749..8770
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8933..9037
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9038..9106,9107..9898)
                     /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(9256..9275)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10069..10657
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10558..10577
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10811..10828
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10945..10966
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10981..10986,10987..11004,11005..11011)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11019..11035
                     /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     join(11024..11038,1..8)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
       61 gaacaaaagc tggtactatt agctgaattg ccactgctat cgttgttagt ggcgttagtg
      121 cttgcattca aagacatgga gggcgttatt acgccggagc tcctcgacag cagatctgat
      181 gactggtcaa tatatttttg cattgaggct ctgtttggaa ttatattttg agatgaccca
      241 tctaatgtac tggtatcacc agatttcatg tcgtttttta aagcggctgc ttgagtctta
      301 gcaatagcgt caccatctgg tgaatccttt gaaggaacca ctgacgaagg tttggacagt
      361 gacgaagagg atctttcctg ctttgaatta gtcgcgctgg gagcagatga cgagttggtg
      421 gagctggggg caggattgct ggccgtcgtg ggtcctgaat gggtccttgg ctggtccatc
      481 tctattctga aaacggaaga ggagtaggga atattactgg ctgaaaataa gtcttgaatg
      541 aacgtatacg cgtatatttc taccaatctc tcaacactga gtaatggtag ttataagaaa
      601 gagaccgagt tagggacagt tagaggcggt ggagatattc cttatggcat gtctggcgat
      661 gataaaactt ttcaaacggc agccccgatc taaaagagct gacagggaaa tggtcagaaa
      721 aagaaacgtg cacccgcccg tctggacgcg ccgctcaccc gcacggcaga gaccaatcag
      781 taaaaatcaa cggttaacga cattactata tatataatat aggaagcatt taatagaaca
      841 gcatcgtaat atatgtgtac tttgcagtta tgacgccaga tggcagtagt ggaagatatt
      901 ctttattgaa aaatagcttg tcaccttacg tacaatcttg atccggagct tttctttttt
      961 tgccgattaa gaattcggtc gaaaaaagaa aaggagaggg ccaagaggga gggcattggt
     1021 gactattgag cacgtgagta tacgtgatta agcacacaaa ggcagcttgg agtatgctcg
     1081 ttaaagtgtg tggtttgcaa accgttgaag ctgcaaagac tgctgtggat gatggtgctg
     1141 attacttagg tatcatttgt gttcccggta ggaaaagaac cattgactca tctgttgcga
     1201 aaggtatttc aactgcagtt caccaacaag agaacgtgaa aggtactaaa ctagtcgggg
     1261 tgtttagaaa tcagtccgtt gatgatgtcc ttcaactgta ccacgaatat aatctagatg
     1321 tgatacaatt acatggagat gaagatatta aagaatacag atctttgatc ccatcttcaa
     1381 ttccaatcat taagaggttc cagttcccac aggattgtga attactactg gacctgtatg
     1441 aacacgtaga caatgtgctg acgttgttcg attctggtga aggtggcact ggtgagaaat
     1501 tgaattggag tgcaatttcc agttggtctg caagtcatcc cgagataaaa ttcattatcg
     1561 ctggtggatt gaatcctgat aacgtttctg ttgccattaa tatgttacca aatgcgatcg
     1621 gtgtcgatgt aagtggagga gtagagactg atggtatcaa ggatttagaa aaggtaaagc
     1681 tattcatcca gcaggcctct caatagagga aaaaacgtat ctttcttcaa ttacacatac
     1741 gtatatataa attgatatat atatagtgaa tgatcgttcc gcggccgctc tagggtaccg
     1801 ggccccccct cgaggtcgac ggtatcgata agcttgaatt cttattacga tcctcgcgcc
     1861 ccctacccac cgtactcgtc aattccaagg gcatcggtaa acatctgctc aaactcgaag
     1921 tcggccatat ccagagcgcc gtagggggcg gagtcgtggg gggtaaatcc cggacccggg
     1981 gaatccccgt cccccaacat gtccagatcg aaatcgtcta gcgcgtcggc atgcgccatc
     2041 gccacgtcct cgccgtctaa gtggagctcg tcccccaggc tgacatcggt cgggggggcc
     2101 gtcgacagtc tgcgcgtgtg tcccgcgggg agaaaggaca ggcgcggagc cgccagcccc
     2161 gcctcttcgg gggcgtcgtc gtccgggaga tcgagcaggc cctcgatggt agacccgtaa
     2221 ttgtttttcg tacgcgcgcg gctgtacgcg gacccacttt cacatttaag ttgtttttct
     2281 aatccgcata tgatcaattc aaggccgaat aagaaggctg gctctgcgcc ttggtgatca
     2341 aataattcga tagcttgtcg taataatggc ggcatactat cagtagtagg tgtttccctt
     2401 tcttctttag cgacttgatg ctcttgatct tccaatacgc aacctaaagt aaaatgcccc
     2461 acagcgctga gtgcatataa tgcattctct agtgaaaaac cttgttggca taaaaaggct
     2521 aattgatttt cgagagtttc atactgtttt tctgtaggcc gtgtacctaa atgtactttt
     2581 gctccatcgc gatgacttag taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa
     2641 tcttgccagc tttccccttc taaagggcaa aagtgagtat ggtgcctatc taacatctca
     2701 atggctaagg cgtcgagcaa agcccgctta ttttttacat gccaatacaa tgtaggctgc
     2761 tctacaccta gcttctgggc gagtttacgg gttgttaaac cttcgattcc gacctcatta
     2821 agcagctcta atgcgctgtt aatcacttta cttttatcta atctagacat atgaattaat
     2881 tcggggccgc ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa acagcgtgga
     2941 tggcgtctcc aggcgatctg acggttcact aaacgagctc tgcttatata gacctcccac
     3001 cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag
     3061 tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga
     3121 aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcacc
     3181 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
     3241 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
     3301 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     3361 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     3421 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     3481 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa
     3541 ctagtcaata atcaatgtca acatggcggt aatgttggac atgagccaat ataaatgtac
     3601 atattatgat atggatacaa cgtatgcaat gggccaagct cctcgagtaa ttcgcgccac
     3661 ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa
     3721 aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt
     3781 gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg tcgctcttat
     3841 tgaccacacc tctaccggca gatccgctag ggataacagg gtaatataga tcaattcctc
     3901 gatcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
     3961 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
     4021 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
     4081 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata
     4141 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
     4201 gctcggtacc ctatggcatg catgtgctct gtatgtatat aaaactcttg ttttcttctt
     4261 ttctctaaat attctttcct tatacattag gtcctttgta gcataaatta ctatacttct
     4321 atagacacgc aaacacaaat acacacacta aattaccgga tcaattcggg ggatccgttt
     4381 aaacaggcct gttaacatcg atagcggcga tggtgtctaa gggcgaagag ctgatcaagg
     4441 aaaatatgcg tatgaaggtg gtcatggaag gttcggtcaa cggccaccaa ttcaaatgca
     4501 caggtgaagg agaaggcaga ccgtacgagg gagtgcaaac catgaggatc aaagtcatcg
     4561 agggaggacc cctgccattt gcctttgaca ttcttgccac gtcgttcatg tatggcagcc
     4621 gtacctttat caagtacccg gccgacatcc ctgatttctt taaacagtcc tttcctgagg
     4681 gttttacttg ggaaagagtt acgagatacg aagatggtgg agtcgtcacc gtcacgcagg
     4741 acaccagcct tgaggatggc gagctcgtct acaacgtcaa ggtcagaggg gtaaactttc
     4801 cctccaatgg tcccgtgatg cagaagaaga ccaagggttg ggagcctaat acagagatga
     4861 tgtatccagc agatggtggt ctgagaggat acactgacat cgcactgaaa gttgatggtg
     4921 gtggccatct gcactgcaac ttcgtgacaa cttacaggtc aaaaaagacc gtcgggaaca
     4981 tcaagatgcc cggtgtccat gccgttgatc accgcctgga aaggatcgag gagagtgaca
     5041 atgaaaccta cgtagtgcaa agagaagtgg cagttgccaa atacagcaac cttggtggtg
     5101 gcatggacga gctgtacaag ggagccggaa caagtgacat gatgactcac acattaccaa
     5161 gcgaacagac acgtcttgtt cctggatctg actcgagctc tcgtcctaaa aagagacgga
     5221 tctcgaagag atcaaagata atagtatcca ctgtggtctg tattggtttg ttgctggttt
     5281 tagtgcaact ggcatttcca agcagttttg cattacgttc tgcatcacac aagaagaaga
     5341 atgtcatatt cttcgtgacg gatgggatgg gacctgcgtc tttgtccatg gcaagatcat
     5401 tcaaccagca tgttaatgat ttacctatag acgacatcct gacacttgac gagcatttta
     5461 tcgggtcttc gagaacaagg tcctcggact cactagtcac tgactcagct gcgggcgcca
     5521 ccgcatttgc atgtgcatta aagagctata atggggccat cggtgttgat ccgcatcaca
     5581 gaccttgtgg tacagtgctc gaggcggcta agttagcagg ttatctcact gggcttgttg
     5641 ttacgacaag aatcacagat gccactccgg cttcattcag ttcacatgtt gattacagat
     5701 ggcaggaaga tcttatcgct acgcaccagt taggcgaata tcctctgggg agagtagtag
     5761 acttactcat gggtggcggc aggagccatt tctaccctca aggcgaaaag gcttctccat
     5821 atggtcacca cggtgctaga aaagatggaa gagatttaat cgatgaggct caatctaatg
     5881 gctggcagta cgtgggtgac cgtaaaaatt ttgattcttt gttgaaaagc cacggtgaaa
     5941 acgttacttt acctttcttg ggtctctttg cagataacga cattcctttt gaaattgaca
     6001 gagatgaaaa ggagtatcct tcccttaagg aacaagtcaa ggtggctttg ggtgccttgg
     6061 aaaaagcttc caatgaggac aaagattcta atgggttttt cttaatggtg gaaggctcca
     6121 gaattgatca tgctggtcac caaaacgatc cagcttcgca ggtaagggag gtgttggctt
     6181 tcgatgaagc attccaatac gttttggagt ttgctgaaaa ttccgatacg gagactgttt
     6241 tggtctctac gtcggatcat gaaacgggtg gtttagtcac ttcaagacaa gtcactgcta
     6301 gctatcctca gtatgtctgg tatccacagg tgctcgccaa cgccacccat tcaggtgagt
     6361 ttttgaaaag aaaactagta gattttgttc atgagcataa gggcgcaagc agcaaaatag
     6421 aaaatttcat caaacatgag atcctggaaa aagatttggg tatttacgat tacacggata
     6481 gcgatttaga aacattaatt catttggatg acaacgcaaa tgctatccaa gacaaactga
     6541 atgacatggt atcatttagg gcccaaattg gctggaccac gcatggtcac agcgcagtcg
     6601 acgtcaacat atatgcatac gccaataaga aagctacatg gtcatatgtc ttgaacaatt
     6661 tacaaggcaa tcacgaaaat acggaagttg gccagttctt agaaaacttt ttagaattga
     6721 acctgaacga agtcactgat ttgattaggg acaccaaaca cacatcagat tttgatgcta
     6781 cagaaatagc tagtgaagta caacattatg acgaatacta ccatgagttg accaactgat
     6841 cctatctaac tctccctctt ttttcacata ttatttaata cgttataatt tatttgaata
     6901 tatatatata tatatatgta tatatactaa ccgcatgaca cgtactgtat cgcatttcgt
     6961 ttcggagtgc aggcgagttt taggtgcatt ctatagcgtg aagaagtgaa atctcgtcat
     7021 ttcttcaagt tatgccattc agtccagatt gtttcacatg ctttcatata gtcatttttg
     7081 tcgtcgcgaa cttcaaattt gaaaacatca tcatacagag agatttctcc aataatgatc
     7141 ctttctttgt cgttgtcatc atcggcatcg tcgttaagtg aagcgccctc agcaagcttg
     7201 atatcgaatt cctgcagccc gggggatcca ctagttctag agcggccgcc accgcggtgg
     7261 gttattactg agtagtattt atttaagtat tgtttgtgca cttgcctgca ggccttttga
     7321 aaagcaagca taaaagatct aaacataaaa tctgtaaaat aacaagatgt aaagataatg
     7381 ctaaatcatt tggctttttg attgattgta caggaaaata tacatcgcag ggggttgact
     7441 tttaccattt caccgcaatg gaatcaaact tgttgaagag aatgttcaca ggcgcatacg
     7501 ctacaatgac ccgattcttg ctagcctttt ctcggtcttg caaacaaccg ccggcagctt
     7561 agtatataaa tacacatgta catacctctc tccgtatcct cgtaatcatt ttcttgtatt
     7621 tatcgtcttt tcgctgtaaa aactttatca cacttatctc aaatacactt attaaccgct
     7681 tttactatta tcttctacgc tgacagtaat atcaaacagt gacacatatt aaacacagtg
     7741 gtttctttgc ataaacacca tcagcctcaa gtcgtcaagt aaagatttcg tgttcatgca
     7801 gatagataac aatctatatg ttgataatta gcgttgcctc atcaatgcga gatccgttta
     7861 accggaccct agtgcactta ccccacgttc ggtccactgt gtgccgaaca tgctccttca
     7921 ctattttaac atgtggaatt cttgaaagaa tgaaatcgcc atgccaagcc atcacacggt
     7981 cttttatgca attgattgac cgcctgcaac acataggcag taaaattttt actgaaacgt
     8041 atataatcat cataagcgac aagtgaggca acacctttgt taccacattg acaaccccag
     8101 gtattcatac ttcctattag cggaatcagg agtgcaaaaa gagaaaataa aagtaaaaag
     8161 gtagggcaac acatagtatg aatacaaacg ttccaatatt cagttctccg gtcagagatt
     8221 taccaaggtc tttcgaacaa aaacatttag cggttgtaga caattcgccc tatagtgagt
     8281 cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     8341 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     8401 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     8461 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     8521 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     8581 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     8641 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     8701 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     8761 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     8821 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     8881 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     8941 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     9001 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     9061 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     9121 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     9181 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     9241 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     9301 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     9361 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     9421 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     9481 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     9541 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     9601 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     9661 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     9721 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     9781 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     9841 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     9901 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     9961 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
    10021 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
    10081 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
    10141 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
    10201 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
    10261 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
    10321 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
    10381 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
    10441 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
    10501 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
    10561 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
    10621 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
    10681 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
    10741 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
    10801 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
    10861 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
    10921 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
    10981 tttacacttt atgctcccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaattt
//
