LOCUS       sequence-235890-       13965 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-inducible expression of IDH1 in mammalian cells, 3rd generation 
            lentiviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13965)
  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 13965)
  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-FLAG
FEATURES             Location/Qualifiers
     source          1..13965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    2614..2638
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        2670..2984
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    2678..2696
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2714..2732
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2749..2767
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2785..2803
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2821..2839
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2856..2874
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2892..2910
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2957..2981
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             4260..4325
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(4346..4370)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        4420..5631
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5468..5487
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(5756..6373,6374..6379,6380..6415,6416..6418,6419..6454,
                     6455..6457,6458..6493,6494..6502)
                     /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    6647..7109
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6703..6720)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6930..6949
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7168..8187
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             7168..8187
                     /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    8653..9241
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8706..8726)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9124..9135)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9244..9260)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9245..9261)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9770..9950
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9976..9993)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9982..10206
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10252..10680
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10339..10358)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10549..10570
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10689..10709)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10694..11023
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10874..11009
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10936..10955
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11071..11118
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11137..11511
                     /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    11641..11762
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11678..11697)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11732..11751
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11811..11827)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11811..11827)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11824..11846)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11835..11851
                     /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(11859..11865,11866..11883,11884..11889))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11904..11925
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12042..12059)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12213..12801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12293..12312)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12972..13763,13764..13832))
                     /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     13595..13614
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13833..13937)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaattc tagacagtta tcaacaagtt tgtacaaaaa agcaggcttt
     2641 aaaggaacca attcagtcga ctggatcccg agtttactcc ctatcagtga tagagaacgt
     2701 atgtcgagtt tactccctat cagtgataga gaacgatgtc gagtttactc cctatcagtg
     2761 atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttac
     2821 tccctatcag tgatagagaa cgtatgtcga gtttatccct atcagtgata gagaacgtat
     2881 gtcgagttta ctccctatca gtgatagaga acgtatgtcg aggtaggcgt gtacggtggg
     2941 aggcctatat aagcagagct cgtttagtga accgtcagat cgccactagt gccaccatgt
     3001 ccaaaaaaat cagtggcggt tctgtggtag agatgcaagg agatgaaatg acacgaatca
     3061 tttgggaatt gattaaagag aaactcattt ttccctacgt ggaattggat ctacatagct
     3121 atgatttagg catagagaat cgtgatgcca ccaacgacca agtcaccaag gatgctgcag
     3181 aagctataaa gaagcataat gttggcgtca aatgtgccac tatcactcct gatgagaaga
     3241 gggttgagga gttcaagttg aaacaaatgt ggaaatcacc aaatggcacc atacgaaata
     3301 ttctgggtgg cacggtcttc agagaagcca ttatctgcaa aaatatcccc cggcttgtga
     3361 gtggatgggt aaaacctatc atcataggtc gtcatgctta tggggatcaa tacagagcaa
     3421 ctgattttgt tgttcctggg cctggaaaag tagagataac ctacacacca agtgacggaa
     3481 cccaaaaggt gacatacctg gtacataact ttgaagaagg tggtggtgtt gccatgggga
     3541 tgtataatca agataagtca attgaagatt ttgcacacag ttccttccaa atggctctgt
     3601 ctaagggttg gcctttgtat ctgagcacca aaaacactat tctgaagaaa tatgatgggc
     3661 gttttaaaga catctttcag gagatatatg acaagcagta caagtcccag tttgaagctc
     3721 aaaagatctg gtatgagcat aggctcatcg acgacatggt ggcccaagct atgaaatcag
     3781 agggaggctt catctgggcc tgtaaaaact atgatggtga cgtgcagtcg gactctgtgg
     3841 cccaagggta tggctctctc ggcatgatga ccagcgtgct ggtttgtcca gatggcaaga
     3901 cagtagaagc agaggctgcc cacgggactg taacccgtca ctaccgcatg taccagaaag
     3961 gacaggagac gtccaccaat cccattgctt ccatttttgc ctggaccaga gggttagccc
     4021 acagagcaaa gcttgataac aataaagagc ttgccttctt tgcaaatgct ttggaagaag
     4081 tctctattga gacaattgag gctggcttca tgaccaagga cttggctgct tgcattaaag
     4141 gtttacccaa tgtgcaacgt tctgactact tgaatacatt tgagttcatg gataaacttg
     4201 gagaaaactt gaagatcaaa ctagctcagg ccaaacttta ttctagagga tcccgggctg
     4261 actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg
     4321 acaagtagtg actcgagata tctagaccca gctttcttgt acaaagtggt tgataacatc
     4381 tcgagccata aagatggtta attaacccac ccaagatctg gcctccgcgc cgggttttgg
     4441 cgcctcccgc gggcgccccc ctcctcacgg cgagcgctgc cacgtcagac gaagggcgca
     4501 gcgagcgtcc tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact
     4561 cggccttaga accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta
     4621 gggcactggt tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg
     4681 attctgcgga gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg
     4741 gtgtggcaca gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc
     4801 gctgtgatcg tcacttggtg agtagcgggc tgctgggctg gccggggctt tcgtggccgc
     4861 cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc
     4921 gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg cggctgttcc
     4981 cgagtcttga atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg
     5041 ggcatggtgg gcggcaagaa cccaaggtct tgaggccttc gctaatgcgg gaaagctctt
     5101 attcgggtga gatgggctgg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac
     5161 tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag
     5221 tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg
     5281 gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc
     5341 aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg
     5401 tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt
     5461 aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa
     5521 gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag
     5581 actagtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga cgaagcttgg
     5641 taccgagctc ggatctccac cccgtaccgg tcctgcagga tttaaatggc gcgccgctag
     5701 cgctaccgga ctcagatctc gagctcaagc ttcgaattct gcagtcgaat tcaccatgtc
     5761 tagactggac aagagcaaag tcataaacgg agctctggaa ttactcaatg gtgtcggtat
     5821 cgaaggcctg acgacaagga aactcgctca aaagctggga gttgagcagc ctaccctgta
     5881 ctggcacgtg aagaacaagc gggccctgct cgatgccctg ccaatcgaga tgctggacag
     5941 gcatcatacc cacttctgcc ccctggaagg cgagtcatgg caagactttc tgcggaacaa
     6001 cgccaagtca taccgctgtg ctctcctctc acatcgcgac ggggctaaag tgcatctcgg
     6061 cacccgccca acagagaaac agtacgaaac cctggaaaat cagctcgcgt tcctgtgtca
     6121 gcaaggcttc tccctggaga acgcactgta cgctctgtcc gccgtgggcc actttacact
     6181 gggctgcgta ttggaggaac aggagcatca agtagcaaaa gaggaaagag agacacctac
     6241 caccgattct atgcccccac ttctgagaca agcaattgag ctgttcgacc ggcagggagc
     6301 cgaacctgcc ttccttttcg gcctggaact aatcatatgt ggcctggaga aacagctaaa
     6361 gtgcgaaagc ggcgggccga ccgacgccct tgacgatttt gacttagaca tgctcccagc
     6421 cgatgccctt gacgactttg accttgatat gctgcctgct gacgctcttg acgattttga
     6481 ccttgacatg ctccccgggt aactaagtaa ggatccgcgg ccgcactaga ggaattccgc
     6541 ccctctcccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     6601 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     6661 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     6721 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     6781 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     6841 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     6901 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     6961 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     7021 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     7081 tggttttcct ttgaaaaaca cgatgataag cttaccggta cgcgtgatag cttgccacaa
     7141 cccgtaccaa agatggatag atccggaaag cctgaactca ccgcgacgtc tgtcgagaag
     7201 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     7261 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     7321 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
     7381 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc
     7441 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
     7501 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
     7561 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
     7621 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
     7681 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
     7741 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
     7801 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc
     7861 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
     7921 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa
     7981 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
     8041 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
     8101 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc
     8161 agcactcgtc cgagggcaaa ggaatagagt agatgccgac cgaacaagag ctgatttcga
     8221 gaacgcctca gccagcaact cgcgcgagcc tagcaaggca aatgcgagag aacggcctta
     8281 cgcttggtgg cacagttctc gtccacagtt cgctaagctc gctcggctgg gtcgcgggag
     8341 ggccggtcgc agtgattcag gcccttctgg attgtgttgg tccccagggc acgattgtca
     8401 tgcccacgca ctcgggtgat ctgactgatc ccgcagattg gagatcgccg cccgtgcctg
     8461 ccgattgggt gcagatctct agatctgtac aagtaaagcg gccgcgactc tagatcataa
     8521 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     8581 tgaacctgaa acataaaatg aatgcaattg ttgttgttta gtccctccca attcgatatc
     8641 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     8701 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     8761 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     8821 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     8881 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     8941 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     9001 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     9061 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     9121 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     9181 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     9241 catcgatacc gtcgacctcg atcgagacct agaaaaacat ggagcaatca caagtagcaa
     9301 tacagcagct accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg
     9361 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
     9421 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     9481 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt ggcagaacta
     9541 cacaccaggg ccagggatca gatatccact gacctttgga tggtgctaca agctagtacc
     9601 agttgagcaa gagaaggtag aagaagccaa tgaaggagag aacacccgct tgttacaccc
     9661 tgtgagcctg catgggatgg atgacccgga gagagaagta ttagagtgga ggtttgacag
     9721 ccgcctagca tttcatcaca tggcccgaga gctgcatccg gactgtactg ggtctctctg
     9781 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     9841 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     9901 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gggcccgttt
     9961 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
    10021 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
    10081 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
    10141 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
    10201 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
    10261 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
    10321 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
    10381 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
    10441 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
    10501 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
    10561 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
    10621 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
    10681 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
    10741 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
    10801 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
    10861 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
    10921 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
    10981 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
    11041 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc
    11101 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt
    11161 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg
    11221 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg
    11281 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg
    11341 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac
    11401 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg
    11461 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg acacgtgcta
    11521 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
    11581 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
    11641 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
    11701 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
    11761 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
    11821 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
    11881 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
    11941 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
    12001 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
    12061 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
    12121 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
    12181 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
    12241 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
    12301 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
    12361 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
    12421 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
    12481 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
    12541 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
    12601 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
    12661 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
    12721 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
    12781 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
    12841 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
    12901 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
    12961 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
    13021 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
    13081 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
    13141 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
    13201 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
    13261 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
    13321 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
    13381 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
    13441 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
    13501 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
    13561 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
    13621 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
    13681 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
    13741 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
    13801 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
    13861 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
    13921 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgac
//
