LOCUS       sequence-445477-       14098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-inducible expression of mutant IDH2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14098)
  AUTHORS   Lewis CA, Parker SJ, Fiske BP, McCloskey D, Gui DY, Green CR, Vokes 
            NI, Feist AM, Vander Heiden MG, Metallo CM
  TITLE     Tracing compartmentalized NADPH metabolism in the cytosol and 
            mitochondria of mammalian cells.
  JOURNAL   Mol Cell. 2014 Jul 17;55(2):253-63. doi: 
            10.1016/j.molcel.2014.05.008. Epub 2014 May 29.
  PUBMED    24882210
REFERENCE   2  (bases 1 to 14098)
  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.2014.05.008"; journalName: "Mol Cell"; date: 
            "2014-07-17- 17"; volume: "55"; issue: "2"; pages: "253-63"
COMMENT     Sequence Label: pSLIK-IDH2-R172K-FLAG
FEATURES             Location/Qualifiers
     source          1..14098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..315
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    9..27
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    45..63
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    80..98
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    116..134
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    152..170
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    187..205
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    223..241
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     288..312
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      322..331
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1705..1770
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(1791..1815)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        1865..3076
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2913..2932
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(3201..3818,3819..3824,3825..3860,3861..3863,3864..3899,
                     3900..3902,3903..3938,3939..3947)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    4000..4573
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4167..4184)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4394..4413
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4632..5651
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             4632..5651
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     misc_feature    6117..6705
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6170..6190)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6588..6599)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6708..6724)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6709..6725)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7234..7414
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7440..7457)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7446..7670
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7716..8144
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7803..7822)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8013..8034
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8153..8173)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8158..8487
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8338..8473
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8400..8419
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8535..8582
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8601..8975
                     /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    9105..9226
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9142..9161)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9196..9215
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9275..9291)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9275..9291)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9288..9310)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9299..9315
                     /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(9323..9329,9330..9347,9348..9353))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9368..9389
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9506..9523)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9677..10265)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9757..9776)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10436..11227,11228..11296))
                     /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     11059..11078
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11297..11401)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(11476..11495)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        11667..12046
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        12048..12246
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             12264..12444
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    12491..12616
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    13109..13342
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             13527..13571
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    13869..13986
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    14043..14067
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 gagtttactc cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag
       61 agaacgatgt cgagtttact ccctatcagt gatagagaac gtatgtcgag tttactccct
      121 atcagtgata gagaacgtat gtcgagttta ctccctatca gtgatagaga acgtatgtcg
      181 agtttatccc tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag
      241 aacgtatgtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg
      301 aaccgtcaga tcgccactag tgccaccatg gccggctacc tgcgggtcgt gcgctcgctc
      361 tgcagagcct caggctcgcg gccggcctgg gcgccggcgg ccctgacagc ccccacctcg
      421 caagagcagc cgcggcgcca ctatgccgac aaaaggatca aggtggcgaa gcccgtggtg
      481 gagatggatg gtgatgagat gacccgtatt atctggcagt tcatcaagga gaagctcatc
      541 ctgccccacg tggacatcca gctaaagtat tttgacctcg ggctcccaaa ccgtgaccag
      601 actgatgacc aggtcaccat tgactctgca ctggccaccc agaagtacag tgtggctgtc
      661 aagtgtgcca ccatcacccc tgatgaggcc cgtgtggaag agttcaagct gaagaagatg
      721 tggaaaagtc ccaatggaac tatccggaac atcctggggg ggactgtctt ccgggagccc
      781 atcatctgca aaaacatccc acgcctagtc cctggctgga ccaagcccat caccattggc
      841 aagcacgccc atggcgacca gtacaaggcc acagactttg tggcagaccg ggccggcact
      901 ttcaaaatgg tcttcacccc aaaagatggc agtggtgtca aggagtggga agtgtacaac
      961 ttccccgcag gcggcgtggg catgggcatg tacaacaccg acgagtccat ctcaggtttt
     1021 gcgcacagct gcttccagta tgccatccag aagaaatggc cgctgtacat gagcaccaag
     1081 aacaccatac tgaaagccta cgatgggcgt ttcaaggaca tcttccagga gatctttgac
     1141 aagcactata agaccgactt cgacaagaat aagatctggt atgagcaccg gctcattgat
     1201 gacatggtgg ctcaggtcct caagtcttcg ggtggctttg tgtgggcctg caagaactat
     1261 gacggagatg tgcagtcaga catcctggcc cagggctttg gctcccttgg cctgatgacg
     1321 tccgtcctgg tctgccctga tgggaagacg attgaggctg aggccgctca tgggaccgtc
     1381 acccgccact atcgggagca ccagaagggc cggcccacca gcaccaaccc catcgccagc
     1441 atctttgcct ggacacgtgg cctggagcac cgggggaagc tggatgggaa ccaagacctc
     1501 atcaggtttg cccagatgct ggagaaggtg tgcgtggaga cggtggagag tggagccatg
     1561 accaaggacc tggcgggctg cattcacggc ctcagcaatg tgaagctgaa cgagcacttc
     1621 ctgaacacca cggacttcct cgacaccatc aagagcaacc tggacagagc cctgggcagg
     1681 cagtattcta gaggatcccg ggctgactac aaagaccatg acggtgatta taaagatcat
     1741 gacatcgact acaaggatga cgatgacaag tagtgactcg agatatctag acccagcttt
     1801 cttgtacaaa gtggttgata acatctcgag ccataaagat ggttaattaa cccacccaag
     1861 atctggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcct cacggcgagc
     1921 gctgccacgt cagacgaagg gcgcagcgag cgtcctgatc cttccgcccg gacgctcagg
     1981 acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt
     2041 ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc
     2101 gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc
     2161 cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt
     2221 gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg
     2281 ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg tgtggagaga
     2341 ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg ggttgggggg
     2401 agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt gaggcgggct
     2461 gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa ggtcttgagg
     2521 ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctggggcac catctgggga
     2581 ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg
     2641 cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg
     2701 tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg
     2761 tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct
     2821 gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg
     2881 gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc
     2941 ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg
     3001 gtcaatatgt aattttcagt gttagactag taaattgtcc gctaaattct ggccgttttt
     3061 ggcttttttg ttagacgaag cttggtaccg agctcggatc tccaccccgt accggtcctg
     3121 caggatttaa atggcgcgcc gctagcgcta ccggactcag atctcgagct caagcttcga
     3181 attctgcagt cgaattcacc atgtctagac tggacaagag caaagtcata aacggagctc
     3241 tggaattact caatggtgtc ggtatcgaag gcctgacgac aaggaaactc gctcaaaagc
     3301 tgggagttga gcagcctacc ctgtactggc acgtgaagaa caagcgggcc ctgctcgatg
     3361 ccctgccaat cgagatgctg gacaggcatc atacccactt ctgccccctg gaaggcgagt
     3421 catggcaaga ctttctgcgg aacaacgcca agtcataccg ctgtgctctc ctctcacatc
     3481 gcgacggggc taaagtgcat ctcggcaccc gcccaacaga gaaacagtac gaaaccctgg
     3541 aaaatcagct cgcgttcctg tgtcagcaag gcttctccct ggagaacgca ctgtacgctc
     3601 tgtccgccgt gggccacttt acactgggct gcgtattgga ggaacaggag catcaagtag
     3661 caaaagagga aagagagaca cctaccaccg attctatgcc cccacttctg agacaagcaa
     3721 ttgagctgtt cgaccggcag ggagccgaac ctgccttcct tttcggcctg gaactaatca
     3781 tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg gccgaccgac gcccttgacg
     3841 attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt gatatgctgc
     3901 ctgctgacgc tcttgacgat tttgaccttg acatgctccc cgggtaacta agtaaggatc
     3961 cgcggccgca ctagaggaat tccgcccctc tccccccccc ccctctccct cccccccccc
     4021 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     4081 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     4141 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     4201 cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct
     4261 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     4321 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     4381 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     4441 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     4501 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     4561 aacacgatga taagcttacc ggtacgcgtg atagcttgcc acaacccgta ccaaagatgg
     4621 atagatccgg aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     4681 tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct
     4741 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     4801 aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg
     4861 acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     4921 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca
     4981 tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
     5041 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
     5101 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
     5161 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
     5221 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
     5281 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
     5341 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat
     5401 cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
     5461 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
     5521 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg
     5581 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
     5641 caaaggaata gagtagatgc cgaccgaaca agagctgatt tcgagaacgc ctcagccagc
     5701 aactcgcgcg agcctagcaa ggcaaatgcg agagaacggc cttacgcttg gtggcacagt
     5761 tctcgtccac agttcgctaa gctcgctcgg ctgggtcgcg ggagggccgg tcgcagtgat
     5821 tcaggccctt ctggattgtg ttggtcccca gggcacgatt gtcatgccca cgcactcggg
     5881 tgatctgact gatcccgcag attggagatc gccgcccgtg cctgccgatt gggtgcagat
     5941 ctctagatct gtacaagtaa agcggccgcg actctagatc ataatcagcc ataccacatt
     6001 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     6061 aatgaatgca attgttgttg tttagtccct cccaattcga tatcaagctt atcgataatc
     6121 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     6181 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     6241 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     6301 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     6361 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     6421 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     6481 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     6541 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     6601 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     6661 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac
     6721 ctcgatcgag acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat
     6781 gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct
     6841 caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     6901 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg
     6961 tggatctacc acacacaagg ctacttccct gattggcaga actacacacc agggccaggg
     7021 atcagatatc cactgacctt tggatggtgc tacaagctag taccagttga gcaagagaag
     7081 gtagaagaag ccaatgaagg agagaacacc cgcttgttac accctgtgag cctgcatggg
     7141 atggatgacc cggagagaga agtattagag tggaggtttg acagccgcct agcatttcat
     7201 cacatggccc gagagctgca tccggactgt actgggtctc tctggttaga ccagatctga
     7261 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     7321 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     7381 agaccctttt agtcagtgtg gaaaatctct agcagggccc gtttaaaccc gctgatcagc
     7441 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     7501 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     7561 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     7621 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     7681 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     7741 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     7801 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     7861 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     7921 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     7981 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     8041 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     8101 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     8161 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     8221 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     8281 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     8341 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     8401 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     8461 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     8521 tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg
     8581 acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc
     8641 gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg
     8701 tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc
     8761 aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc
     8821 tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca
     8881 tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca
     8941 actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca
     9001 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     9061 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     9121 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     9181 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     9241 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     9301 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     9361 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     9421 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     9481 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     9541 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     9601 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     9661 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     9721 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     9781 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     9841 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     9901 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     9961 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
    10021 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
    10081 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
    10141 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
    10201 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
    10261 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
    10321 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
    10381 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
    10441 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
    10501 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
    10561 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
    10621 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
    10681 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
    10741 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
    10801 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
    10861 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
    10921 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
    10981 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
    11041 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
    11101 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
    11161 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
    11221 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
    11281 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
    11341 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
    11401 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
    11461 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
    11521 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
    11581 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
    11641 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
    11701 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
    11761 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
    11821 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
    11881 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
    11941 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
    12001 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
    12061 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
    12121 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
    12181 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
    12241 aagcagcgcg ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc
    12301 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
    12361 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
    12421 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa
    12481 accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga
    12541 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga
    12601 tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg
    12661 gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga
    12721 gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat
    12781 actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa
    12841 tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc
    12901 tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc
    12961 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
    13021 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
    13081 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
    13141 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
    13201 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
    13261 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
    13321 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
    13381 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
    13441 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
    13501 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
    13561 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
    13621 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga
    13681 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg
    13741 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca
    13801 ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc
    13861 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
    13921 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
    13981 aattttcggg tttattacag ggacagcaga gatccagttt ggttaattct agacagttat
    14041 caacaagttt gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac tggatccc
//
