LOCUS       sequence-333648-       13403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EGFP cDNA and KRAS G12V cDNA in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13403)
  AUTHORS   Carver J, Dexheimer TS, Hsu D, Weng MT, Smith JL, Guha R, Jadhav A, 
            Simeonov A, Luo J
  TITLE     A high-throughput assay for small molecule destabilizers of the KRAS
            oncoprotein.
  JOURNAL   PLoS One. 2014 Aug 5;9(8):e103836. doi: 
            10.1371/journal.pone.0103836. eCollection 2014.
  PUBMED    25093678
REFERENCE   2  (bases 1 to 13403)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2014-08-5"; volume: "9(8):e103836. doi"; pages: " 
            10.1371/journal.pone.0103836. eCollection 2014"
COMMENT     Sequence Label: pInducer10b-EGFP-KRAS G12V
FEATURES             Location/Qualifiers
     source          1..13403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    488..721
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             906..950
                     /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    1248..1365
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(1430..1804)
                     /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"
     regulatory      1801..1810
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1814..1832)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1824..1871)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    join(1941..1959,1960..1982,1983..2001,2002..2024,2025..2043,
                     2044..2066,2067..2085,2086..2108,2109..2127,2128..2150,
                     2151..2169,2170..2192,2193..2211)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2244..2282
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2279..2303
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(2375..2377,2378..2380,2381..3091)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2375..2377,2378..2380,2381..3091)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2420..2441)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2681..2700)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3028..3049
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             join(3113..3145,3146..3148,3149..3679)
                     /codon_start=1
                     /product="human oncoprotein generated by the G12V mutation 
                     in the small GTPase K-Ras"
                     /label=K-Ras (G12V)
                     /translation="MTEYKLVVVGAVGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     promoter        3692..4903
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4740..4759
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    4910..4943
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             join(4950..5567,5568..5573,5574..5609,5610..5612,5613..5648,
                     5649..5651,5652..5687,5688..5696)
                     /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    5745..6317
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5912..5929)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6139..6158
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6320..6922
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     6819..6839
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6944..7532
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6997..7017)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7415..7426)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7739..7972
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8073..8090)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8079..8303
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8349..8777
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8436..8455)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8646..8667
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8786..8806)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8791..9120
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8971..9106
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9033..9052
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    9943..10064
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10113..10129)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10113..10129)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10126..10148)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10137..10153
                     /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(10161..10167,10168..10185,10186..10191))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10206..10227
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10344..10361)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10515..11103)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10595..10614)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11274..12065,12066..12134))
                     /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     11897..11916
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12135..12239)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    12287..12421
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             12589..13222
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    13269..13394
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
ORIGIN
        1 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
       61 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
      121 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
      181 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
      241 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
      301 aacaaaagta agaccaccgc acagcaagcg gccggccgct gatcttcaga cctggaggag
      361 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
      421 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
      481 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
      541 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
      601 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
      661 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
      721 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
      781 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
      841 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
      901 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
      961 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     1021 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     1081 caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa
     1141 tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg
     1201 cactgcgtgc gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa
     1261 ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata
     1321 caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg
     1381 gacagcagag atccagtttg gttagtaccg ggcccgctct agtccggaat cagtcctgct
     1441 cctcggccac gaagtgcacg cagttgccgg ccgggtcgcg cagggcgaac tcccgccccc
     1501 acggctgctc gccgatctcg gtcatggccg gcccggaggc gtcccggaag ttcgtggaca
     1561 cgacctccga ccactcggcg tacagctcgt ccaggccgcg cacccacacc caggccaggg
     1621 tgttgtccgg caccacctgg tcctggaccg cgctgatgaa cagggtcacg tcgtcccgga
     1681 ccacaccggc gaagtcgtcc tccacgaagt cccgggagaa cccgagccgg tcggtccaga
     1741 actcgaccgc tccggcgacg tcgcgcgcgg tgagcaccgg aacggcactg gtcaacttgg
     1801 ccatggtggc cctcctatag tgagtcgtat tatactatgc cgatatacta tgccgatgat
     1861 taattgtcaa cacgtgctgc aggtccgagg ttctaggcta gcttaaagga accaattcag
     1921 tcgactggat cctttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
     1981 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
     2041 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
     2101 tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag
     2161 tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa
     2221 gtcgagctcg gtacccgggt cgaggtaggc gtgtacggtg ggaggcctat ataagcagag
     2281 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     2341 gaagacaccg ggaccgatcc agcctaccgg taccatggtg agcaagggcg aggagctgtt
     2401 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     2461 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     2521 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     2581 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     2641 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     2701 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     2761 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     2821 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     2881 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     2941 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     3001 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     3061 gatcactctc ggcatggacg agctgtacaa gggactcgag ggaggatcag gaatgactga
     3121 atataaactt gtggtagttg gagctgttgg cgtaggcaag agtgccttga cgatacagct
     3181 aattcagaat cattttgtgg acgaatatga tccaacaata gaggattcct acaggaagca
     3241 agtagtaatt gatggagaaa cctgtctctt ggatattctc gacacagcag gtcaagagga
     3301 gtacagtgca atgagggacc agtacatgag gactggggag ggctttcttt gtgtatttgc
     3361 cataaataat actaaatcat ttgaagatat tcaccattat agagaacaaa ttaaaagagt
     3421 taaggactct gaagatgtac ctatggtcct agtaggaaat aaatgtgatt tgccttctag
     3481 aacagtagac acaaaacagg ctcaggactt agcaagaagt tatggaattc cttttattga
     3541 aacatcagca aagacaagac agggtgttga tgatgccttc tatacattag ttcgagaaat
     3601 tcgaaaacat aaagaaaaga tgagcaaaga tggtaaaaag aagaaaaaga agtcaaagac
     3661 aaagtgtgta attatgtaac gcgtaagatc tggcctccgc gccgggtttt ggcgcctccc
     3721 gcgggcgccc ccctcctcac ggcgagcgct gccacgtcag acgaagggcg cagcgagcgt
     3781 cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta
     3841 gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg
     3901 gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg
     3961 gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca
     4021 cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat
     4081 cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc
     4141 tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa
     4201 ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt
     4261 gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt
     4321 gggcggcaag aacccaaggt cttgaggcct tcgctaatgc gggaaagctc ttattcgggt
     4381 gagatgggct ggggcaccat ctggggaccc tgacgtgaag tttgtcactg actggagaac
     4441 tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt gccgttgggc agtgcacccg
     4501 tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa
     4561 tgcagggtgg ggccacctgc cggtaggtgt gtggtaggct tttctccgtc gcaggacgca
     4621 gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg ccggacctct ggtgagggga
     4681 gggataagtg aggcgtcagt ttctttggtc ggttttatgt acctatcttc ttaagtagct
     4741 gaagctccgg ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg aagtttttta
     4801 ggcacctttt gaaatgtaat catttgggtc aatatgtaat tttcagtgtt agactagtaa
     4861 attgtccgct aaattctggc cgtttttggc ttttttgtta gacggatcca taacttcgta
     4921 taatgtatgc tatacgaagt tatgccacca tgtctagact ggacaagagc aaagtcataa
     4981 acggagctct ggaattactc aatggtgtcg gtatcgaagg cctgacgaca aggaaactcg
     5041 ctcaaaagct gggagttgag cagcctaccc tgtactggca cgtgaagaac aagcgggccc
     5101 tgctcgatgc cctgccaatc gagatgctgg acaggcatca tacccacttc tgccccctgg
     5161 aaggcgagtc atggcaagac tttctgcgga acaacgccaa gtcataccgc tgtgctctcc
     5221 tctcacatcg cgacggggct aaagtgcatc tcggcacccg cccaacagag aaacagtacg
     5281 aaaccctgga aaatcagctc gcgttcctgt gtcagcaagg cttctccctg gagaacgcac
     5341 tgtacgctct gtccgccgtg ggccacttta cactgggctg cgtattggag gaacaggagc
     5401 atcaagtagc aaaagaggaa agagagacac ctaccaccga ttctatgccc ccacttctga
     5461 gacaagcaat tgagctgttc gaccggcagg gagccgaacc tgccttcctt ttcggcctgg
     5521 aactaatcat atgtggcctg gagaaacagc taaagtgcga aagcggcggg ccgaccgacg
     5581 cccttgacga ttttgactta gacatgctcc cagccgatgc ccttgacgac tttgaccttg
     5641 atatgctgcc tgctgacgct cttgacgatt ttgaccttga catgctcccc gggtaaataa
     5701 cttcgtatag catacattat acgaagtatg gcgcgccaat tccgcccctc tccctccccc
     5761 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     5821 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     5881 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
     5941 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     6001 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     6061 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     6121 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     6181 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt
     6241 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     6301 tgaaaaacac gataatacca tggccaccga gtacaagccc acggtgcgcc tcgccacccg
     6361 cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     6421 gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     6481 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     6541 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggctc
     6601 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     6661 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     6721 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     6781 cgctggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     6841 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     6901 gacccgcaag cccggtgcct gattaattaa gcgcgtctgg aacaatcaac ctctggatta
     6961 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     7021 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     7081 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     7141 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     7201 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     7261 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     7321 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     7381 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     7441 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     7501 gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattctgcag tcgagaccta
     7561 gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg
     7621 ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga
     7681 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa gaggggactg
     7741 gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc
     7801 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     7861 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     7921 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt
     7981 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
     8041 aggccttgac attgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     8101 atctgttgtt tgccccttcc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     8161 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     8221 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     8281 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     8341 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     8401 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     8461 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     8521 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     8581 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     8641 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     8701 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     8761 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     8821 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     8881 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     8941 gtaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     9001 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     9061 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     9121 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgat gaaaaagcct
     9181 gaactcaccg cgacgtgacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata
     9241 tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt
     9301 gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc
     9361 cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata
     9421 acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac
     9481 atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg
     9541 aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt
     9601 gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt
     9661 cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc
     9721 agaagcgcgg ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga
     9781 cgccccagca ctcgtccgag ggcaaaggaa tagcacgtgc tacgagattt cgattccacc
     9841 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     9901 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     9961 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
    10021 ctgcattcta gttgtggttt gtccaaacta atcaatgtat cttatcatgt ctgtataccg
    10081 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
    10141 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
    10201 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
    10261 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
    10321 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
    10381 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
    10441 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
    10501 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
    10561 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
    10621 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
    10681 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
    10741 taggtcgttc gctccaagct gggctgtgtg cactaacccc ccgttcagcc cgaccgctgc
    10801 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
    10861 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
    10921 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
    10981 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
    11041 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
    11101 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
    11161 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
    11221 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
    11281 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
    11341 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
    11401 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
    11461 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
    11521 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
    11581 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
    11641 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
    11701 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
    11761 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
    11821 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
    11881 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
    11941 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
    12001 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
    12061 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
    12121 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
    12181 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
    12241 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatcaact tgtttattgc
    12301 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
    12361 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
    12421 caactggata actcaagcta accaaaatca tcccaaactt cccaccccat accctattac
    12481 cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg aattattttc
    12541 attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagttg gaagggctaa
    12601 ttcactccca aagaagacaa gatatccttg atctgtggat ctaccacaca caaggctact
    12661 tccctgatta gcagaactac acaccagggc caggggtcag atatccactg acctttggat
    12721 ggtgctacaa gctagtacca gttgagccag ataaggtaga agaggccaat aaaggagaga
    12781 acaccagctt gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtgt
    12841 tagagtggag gtttgacagc cgcctagcat ttcatcacgt ggcccgagag ctgcatccgg
    12901 agtacttcaa gaactgctga tatcgagctt gctacaaggg actttccgct ggggactttc
    12961 cagggaggcg tggcctgggc gggactgggg agtggcgagc cctcagatcc tgcatataag
    13021 cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
    13081 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
    13141 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
    13201 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
    13261 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
    13321 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
    13381 ggtgcgagag cgtcagtatt aag
//
