LOCUS       sequence-232105-       13529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-inducible expression of GFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13529)
  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 13529)
  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-GFP
FEATURES             Location/Qualifiers
     source          1..13529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /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    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    2377..2401
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    join(2440..2458,2459..2481,2482..2500,2501..2523,2524..2542,
                     2543..2565,2566..2584,2585..2607,2608..2626,2627..2649,
                     2650..2668,2669..2691,2692..2710)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2743..2781
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2778..2802
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(2888..2890,2891..2893,2894..3607)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2888..2890,2891..2893,2894..3604)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2933..2954)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3194..3213)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3541..3562
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(3660..3684)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        3734..4945
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4782..4801
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(5070..5687,5688..5693,5694..5729,5730..5732,5733..5768,
                     5769..5771,5772..5807,5808..5816)
                     /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    5974..6436
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6030..6047)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6257..6276
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6495..7514
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             6495..7514
                     /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    7980..8568
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8033..8053)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8451..8462)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8571..8587)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8572..8588)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9097..9277
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9303..9320)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9309..9533
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9579..10007
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9666..9685)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9876..9897
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10016..10036)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10021..10350
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10201..10336
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10263..10282
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10398..10445
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10464..10838
                     /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    10968..11089
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11005..11024)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11059..11078
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11138..11154)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11138..11154)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11151..11173)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11162..11178
                     /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(11186..11192,11193..11210,11211..11216))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11231..11252
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11369..11386)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11540..12128)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11620..11639)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12299..13090,13091..13159))
                     /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     12922..12941
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13160..13264)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(13339..13358)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttctagacag ttatcaacaa gtttgtacaa aaaagcaggc
     2401 tttaaaggaa ccaattcagt cgactggatc ctttaccact ccctatcagt gatagagaaa
     2461 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
     2521 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata
     2581 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
     2641 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca
     2701 gtgatagaga aaagtgaaag tcgagctcgg tacccgggtc gaggtaggcg tgtacggtgg
     2761 gaggcctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca
     2821 cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccgcgg ccccgaacta
     2881 gtccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     2941 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     3001 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     3061 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     3121 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     3181 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     3241 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     3301 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     3361 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     3421 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     3481 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     3541 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     3601 caagtaaact agtcgactag ggataacagg gtaattgttt gaatgaggct tcagtctaga
     3661 cccagctttc ttgtacaaag tggttgataa catctcgagc cataaagatg gttaattaac
     3721 ccacccaaga tctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcctc
     3781 acggcgagcg ctgccacgtc agacgaaggg cgcagcgagc gtcctgatcc ttccgcccgg
     3841 acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca gtatcagcag
     3901 aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt tccagagagc
     3961 ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc tccgtggggc
     4021 ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag
     4081 ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc
     4141 gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt
     4201 gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg
     4261 gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg
     4321 aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag
     4381 gtcttgaggc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctggggcacc
     4441 atctggggac cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg
     4501 cgggggcggc agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg
     4561 ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacct
     4621 gccggtaggt gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta
     4681 ggctctcctg aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca
     4741 gtttctttgg tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac
     4801 tatgcgctcg gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta
     4861 atcatttggg tcaatatgta attttcagtg ttagactagt aaattgtccg ctaaattctg
     4921 gccgtttttg gcttttttgt tagacgaagc ttggtaccga gctcggatct ccaccccgta
     4981 ccggtcctgc aggatttaaa tggcgcgccg ctagcgctac cggactcaga tctcgagctc
     5041 aagcttcgaa ttctgcagtc gaattcacca tgtctagact ggacaagagc aaagtcataa
     5101 acggagctct ggaattactc aatggtgtcg gtatcgaagg cctgacgaca aggaaactcg
     5161 ctcaaaagct gggagttgag cagcctaccc tgtactggca cgtgaagaac aagcgggccc
     5221 tgctcgatgc cctgccaatc gagatgctgg acaggcatca tacccacttc tgccccctgg
     5281 aaggcgagtc atggcaagac tttctgcgga acaacgccaa gtcataccgc tgtgctctcc
     5341 tctcacatcg cgacggggct aaagtgcatc tcggcacccg cccaacagag aaacagtacg
     5401 aaaccctgga aaatcagctc gcgttcctgt gtcagcaagg cttctccctg gagaacgcac
     5461 tgtacgctct gtccgccgtg ggccacttta cactgggctg cgtattggag gaacaggagc
     5521 atcaagtagc aaaagaggaa agagagacac ctaccaccga ttctatgccc ccacttctga
     5581 gacaagcaat tgagctgttc gaccggcagg gagccgaacc tgccttcctt ttcggcctgg
     5641 aactaatcat atgtggcctg gagaaacagc taaagtgcga aagcggcggg ccgaccgacg
     5701 cccttgacga ttttgactta gacatgctcc cagccgatgc ccttgacgac tttgaccttg
     5761 atatgctgcc tgctgacgct cttgacgatt ttgaccttga catgctcccc gggtaactaa
     5821 gtaaggatcc gcggccgcac tagaggaatt ccgcccctct cccccccccc cctncccccc
     5881 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     5941 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     6001 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     6061 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     6121 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     6181 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     6241 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     6301 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     6361 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     6421 aaaaacacga tgataagctt accggtacgc gtgatagctt gccacaaccc gtaccaaaga
     6481 tggatagatc cggaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa
     6541 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca
     6601 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
     6661 acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc
     6721 ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg
     6781 tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg
     6841 ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac
     6901 cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc
     6961 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc
     7021 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg
     7081 atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga
     7141 gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt
     7201 ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag
     7261 gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct
     7321 tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc
     7381 gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga
     7441 ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga
     7501 gggcaaagga atagagtaga tgccgaccga acaagagctg atttcgagaa cgcctcagcc
     7561 agcaactcgc gcgagcctag caaggcaaat gcgagagaac ggccttacgc ttggtggcac
     7621 agttctcgtc cacagttcgc taagctcgct cggctgggtc gcgggagggc cggtcgcagt
     7681 gattcaggcc cttctggatt gtgttggtcc ccagggcacg attgtcatgc ccacgcactc
     7741 gggtgatctg actgatcccg cagattggag atcgccgccc gtgcctgccg attgggtgca
     7801 gatctctaga tctgtacaag taaagcggcc gcgactctag atcataatca gccataccac
     7861 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     7921 taaaatgaat gcaattgttg ttgtttagtc cctcccaatt cgatatcaag cttatcgata
     7981 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     8041 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     8101 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     8161 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     8221 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     8281 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     8341 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     8401 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     8461 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     8521 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     8581 gacctcgatc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     8641 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca
     8701 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     8761 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat
     8821 ctgtggatct accacacaca aggctacttc cctgattggc agaactacac accagggcca
     8881 gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag
     8941 aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat
     9001 gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt
     9061 catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc
     9121 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     9181 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     9241 ctcagaccct tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc
     9301 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     9361 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     9421 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     9481 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     9541 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     9601 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     9661 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     9721 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     9781 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     9841 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     9901 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     9961 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
    10021 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
    10081 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
    10141 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
    10201 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
    10261 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
    10321 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
    10381 ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat
    10441 acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg
    10501 cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact
    10561 tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg
    10621 tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg
    10681 agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg
    10741 ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg
    10801 gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt
    10861 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
    10921 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
    10981 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
    11041 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
    11101 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
    11161 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
    11221 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
    11281 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
    11341 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
    11401 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
    11461 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
    11521 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
    11581 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
    11641 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
    11701 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
    11761 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
    11821 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
    11881 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
    11941 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
    12001 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
    12061 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
    12121 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
    12181 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
    12241 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
    12301 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
    12361 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
    12421 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
    12481 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
    12541 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
    12601 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
    12661 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
    12721 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
    12781 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
    12841 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
    12901 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
    12961 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
    13021 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
    13081 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
    13141 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
    13201 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
    13261 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
    13321 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
    13381 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
    13441 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
    13501 tcgcgatgta cgggccagat atacgcgtt
//
