LOCUS       sequence-233142-       13969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-inducible expression of mutant IDH1 in mammalian cells, 3rd 
            generation lentiviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13969)
  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 13969)
  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-IDH1-R132H-FLAG
FEATURES             Location/Qualifiers
     source          1..13969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1981..2000)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2172..2551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2553..2751
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             2769..2949
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2996..3121
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3614..3847
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4032..4076
                     /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    4374..4491
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    4548..4572
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        4604..4918
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    4612..4630
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4648..4666
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4683..4701
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4719..4737
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4755..4773
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4790..4808
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4826..4844
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4891..4915
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             6194..6259
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(6280..6304)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        6354..7565
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     7402..7421
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(7690..8307,8308..8313,8314..8349,8350..8352,8353..8388,
                     8389..8391,8392..8427,8428..8436)
                     /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    8474..9047
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8641..8658)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8868..8887
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             9106..10125
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             9106..10125
                     /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    10591..11179
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10644..10664)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11062..11073)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(11182..11198)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11183..11199)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             11708..11888
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11914..11931)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11920..12144
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      12190..12618
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12277..12296)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12487..12508
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12627..12647)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12632..12961
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12812..12947
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12874..12893
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        13009..13056
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             13075..13449
                     /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    13579..13700
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13616..13635)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13670..13689
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13749..13765)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13749..13765)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13762..13784)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13773..13789
                     /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(13797..13803,13804..13821,13822..13827))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13842..13863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     1981 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     2041 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     2101 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     2161 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     2221 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2281 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     2341 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     2401 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     2461 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     2521 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     2581 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     2641 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     2701 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg
     2761 cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     2821 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     2881 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     2941 tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc
     3001 tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt
     3061 gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt
     3121 cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg
     3181 aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc
     3241 agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca
     3301 accatccctt cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct
     3361 ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag acaagataga
     3421 ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg
     3481 gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa
     3541 ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa
     3601 gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg
     3661 gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc
     3721 agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct
     3781 ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac
     3841 agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga
     3901 atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt
     3961 gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa
     4021 accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga
     4081 attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag
     4141 gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg
     4201 gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg
     4261 aaggaataga agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg
     4321 gatcggcact gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa
     4381 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
     4441 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat
     4501 tacagggaca gcagagatcc agtttggtta attctagaca gttatcaaca agtttgtaca
     4561 aaaaagcagg ctttaaagga accaattcag tcgactggat cccgagttta ctccctatca
     4621 gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacga tgtcgagttt
     4681 actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg
     4741 tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttat ccctatcagt
     4801 gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgaggtag
     4861 gcgtgtacgg tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgccac
     4921 tagtgccacc atgtccaaaa aaatcagtgg cggttctgtg gtagagatgc aaggagatga
     4981 aatgacacga atcatttggg aattgattaa agagaaactc atttttccct acgtggaatt
     5041 ggatctacat agctatgatt taggcataga gaatcgtgat gccaccaacg accaagtcac
     5101 caaggatgct gcagaagcta taaagaagca taatgttggc gtcaaatgtg ccactatcac
     5161 tcctgatgag aagagggttg aggagttcaa gttgaaacaa atgtggaaat caccaaatgg
     5221 caccatacga aatattctgg gtggcacggt cttcagagaa gccattatct gcaaaaatat
     5281 cccccggctt gtgagtggat gggtaaaacc tatcatcata ggtcatcatg cttatgggga
     5341 tcaatacaga gcaactgatt ttgttgttcc tgggcctgga aaagtagaga taacctacac
     5401 accaagtgac ggaacccaaa aggtgacata cctggtacat aactttgaag aaggtggtgg
     5461 tgttgccatg gggatgtata atcaagataa gtcaattgaa gattttgcac acagttcctt
     5521 ccaaatggct ctgtctaagg gttggccttt gtatctgagc accaaaaaca ctattctgaa
     5581 gaaatatgat gggcgtttta aagacatctt tcaggagata tatgacaagc agtacaagtc
     5641 ccagtttgaa gctcaaaaga tctggtatga gcataggctc atcgacgaca tggtggccca
     5701 agctatgaaa tcagagggag gcttcatctg ggcctgtaaa aactatgatg gtgacgtgca
     5761 gtcggactct gtggcccaag ggtatggctc tctcggcatg atgaccagcg tgctggtttg
     5821 tccagatggc aagacagtag aagcagaggc tgcccacggg actgtaaccc gtcactaccg
     5881 catgtaccag aaaggacagg agacgtccac caatcccatt gcttccattt ttgcctggac
     5941 cagagggtta gcccacagag caaagcttga taacaataaa gagcttgcct tctttgcaaa
     6001 tgctttggaa gaagtctcta ttgagacaat tgaggctggc ttcatgacca aggacttggc
     6061 tgcttgcatt aaaggtttac ccaatgtgca acgttctgac tacttgaata catttgagtt
     6121 catggataaa cttggagaaa acttgaagat caaactagct caggccaaac tttattctag
     6181 aggatcccgg gctgactaca aagaccatga cggtgattat aaagatcatg acatcgacta
     6241 caaggatgac gatgacaagt agtgactcga gatatctaga cccagctttc ttgtacaaag
     6301 tggttgataa catctcgagc cataaagatg gttaattaac ccacccaaga tctggcctcc
     6361 gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcctc acggcgagcg ctgccacgtc
     6421 agacgaaggg cgcagcgagc gtcctgatcc ttccgcccgg acgctcagga cagcggcccg
     6481 ctgctcataa gactcggcct tagaacccca gtatcagcag aaggacattt taggacggga
     6541 cttgggtgac tctagggcac tggttttctt tccagagagc ggaacaggcg aggaaaagta
     6601 gtcccttctc ggcgattctg cggagggatc tccgtggggc ggtgaacgcc gatgattata
     6661 taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt
     6721 cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg gctggccggg
     6781 gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt gtggagagac cgccaagggc
     6841 tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg gttgggggga gcgcagcaaa
     6901 atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt
     6961 gaaacaaggt ggggggcatg gtgggcggca agaacccaag gtcttgaggc cttcgctaat
     7021 gcgggaaagc tcttattcgg gtgagatggg ctggggcacc atctggggac cctgacgtga
     7081 agtttgtcac tgactggaga actcggtttg tcgtctgttg cgggggcggc agttatggcg
     7141 gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt
     7201 cacccgttct gttggcttat aatgcagggt ggggccacct gccggtaggt gtgcggtagg
     7261 cttttctccg tcgcaggacg cagggttcgg gcctagggta ggctctcctg aatcgacagg
     7321 cgccggacct ctggtgaggg gagggataag tgaggcgtca gtttctttgg tcggttttat
     7381 gtacctatct tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg gggttggcga
     7441 gtgtgttttg tgaagttttt taggcacctt ttgaaatgta atcatttggg tcaatatgta
     7501 attttcagtg ttagactagt aaattgtccg ctaaattctg gccgtttttg gcttttttgt
     7561 tagacgaagc ttggtaccga gctcggatct ccaccccgta ccggtcctgc aggatttaaa
     7621 tggcgcgccg ctagcgctac cggactcaga tctcgagctc aagcttcgaa ttctgcagtc
     7681 gaattcacca tgtctagact ggacaagagc aaagtcataa acggagctct ggaattactc
     7741 aatggtgtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct gggagttgag
     7801 cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc cctgccaatc
     7861 gagatgctgg acaggcatca tacccacttc tgccccctgg aaggcgagtc atggcaagac
     7921 tttctgcgga acaacgccaa gtcataccgc tgtgctctcc tctcacatcg cgacggggct
     7981 aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga aaatcagctc
     8041 gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct gtccgccgtg
     8101 ggccacttta cactgggctg cgtattggag gaacaggagc atcaagtagc aaaagaggaa
     8161 agagagacac ctaccaccga ttctatgccc ccacttctga gacaagcaat tgagctgttc
     8221 gaccggcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat atgtggcctg
     8281 gagaaacagc taaagtgcga aagcggcggg ccgaccgacg cccttgacga ttttgactta
     8341 gacatgctcc cagccgatgc ccttgacgac tttgaccttg atatgctgcc tgctgacgct
     8401 cttgacgatt ttgaccttga catgctcccc gggtaactaa gtaaggatcc gcggccgcac
     8461 tagaggaatt ccgcccctct cccccccccc cccctaacgt tactggccga agccgcttgg
     8521 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     8581 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     8641 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
     8701 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     8761 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     8821 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     8881 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     8941 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
     9001 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataagct taccggtacg
     9061 cgtgatagct tgccacaacc cgtaccaaag atggatagat ccggaaagcc tgaactcacc
     9121 gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag
     9181 ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc
     9241 ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt
     9301 gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag cgagagcctg
     9361 acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa
     9421 ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt
     9481 agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg
     9541 cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac
     9601 gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac
     9661 tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac
     9721 aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac
     9781 gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc
     9841 tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc
     9901 cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct
     9961 tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca
    10021 caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat
    10081 agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagagtag atgccgaccg
    10141 aacaagagct gatttcgaga acgcctcagc cagcaactcg cgcgagccta gcaaggcaaa
    10201 tgcgagagaa cggccttacg cttggtggca cagttctcgt ccacagttcg ctaagctcgc
    10261 tcggctgggt cgcgggaggg ccggtcgcag tgattcaggc ccttctggat tgtgttggtc
    10321 cccagggcac gattgtcatg cccacgcact cgggtgatct gactgatccc gcagattgga
    10381 gatcgccgcc cgtgcctgcc gattgggtgc agatctctag atctgtacaa gtaaagcggc
    10441 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
    10501 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgtttagt
    10561 ccctcccaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa
    10621 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
    10681 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
    10741 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
    10801 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
    10861 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
    10921 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
    10981 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
    11041 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
    11101 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
    11161 ctttgggccg cctccccgca tcgataccgt cgacctcgat cgagacctag aaaaacatgg
    11221 agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca
    11281 agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta
    11341 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
    11401 tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt
    11461 ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga cctttggatg
    11521 gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg aaggagagaa
    11581 cacccgcttg ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtatt
    11641 agagtggagg tttgacagcc gcctagcatt tcatcacatg gcccgagagc tgcatccgga
    11701 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
    11761 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
    11821 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
    11881 ctctagcagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
    11941 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
    12001 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
    12061 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
    12121 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
    12181 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
    12241 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
    12301 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
    12361 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
    12421 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
    12481 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
    12541 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
    12601 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
    12661 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
    12721 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
    12781 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
    12841 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
    12901 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
    12961 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa
    13021 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc
    13081 aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc
    13141 tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc
    13201 cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc
    13261 ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg
    13321 tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg
    13381 gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag
    13441 caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
    13501 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
    13561 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
    13621 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
    13681 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
    13741 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
    13801 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
    13861 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
    13921 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgct
//
