LOCUS       sequence-423946-       13308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline inducible expression of myc-tagged ERK2-R67S/S153D/D321N
            cDNA in mammilian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13308)
  AUTHORS   Klomp JE, Diehl JN, Klomp JA, Edwards AC, Yang R, Morales AJ, Taylor
            KE, Drizyte-Miller K, Bryant KL, Schaefer A, Johnson JL, Huntsman 
            EM, Yaron TM, Pierobon M, Baldelli E, Prevatte AW, Barker NK, 
            Herring LE, Petricoin EF 3rd, Graves LM, Cantley LC, Cox AD, Der CJ,
            Stalnecker CA
  TITLE     Determining the ERK-regulated phosphoproteome driving KRAS-mutant 
            cancer.
  JOURNAL   Science. 2024 Jun 7;384(6700):eadk0850. doi: 
            10.1126/science.adk0850. Epub 2024 Jun 7.
  PUBMED    38843329
REFERENCE   2  (bases 1 to 13308)
  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: "Science."; 
            date: "2024-06-7"; volume: "384(6700):eadk0850. doi"; pages: " 
            10.1126/science.adk0850. Epub 2024 Jun 7"
COMMENT     Sequence Label: pInducer-myc-ERK2-R67S/S153D/D321N
FEATURES             Location/Qualifiers
     source          1..13308
                     /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"
     protein_bind    join(1983..2001,2002..2024,2025..2043,2044..2066,2067..2085,
                     2086..2108,2109..2127,2128..2150,2151..2169,2170..2192,
                     2193..2211,2212..2234,2235..2253)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    join(2025..2043,2044..2066,2067..2085,2086..2108,2109..2127,
                     2128..2150,2151..2169,2170..2192,2193..2211,2212..2234,
                     2235..2253,2254..2276,2277..2295)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2328..2366
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2363..2387
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2459..2488
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3545..3580
                     /codon_start=1
                     /product="epitope tag derived from MAP kinase 2"
                     /label=Tag-100
                     /translation="EETARFQPGYRS"
     promoter        3597..4808
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4645..4664
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    4815..4848
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             join(4855..5472,5473..5478,5479..5514,5515..5517,5518..5553,
                     5554..5556,5557..5592,5593..5601)
                     /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    5650..6222
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5817..5834)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6044..6063
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6225..6827
                     /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     6724..6744
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6849..7437
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6902..6922)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7320..7331)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7644..7877
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7978..7995)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7984..8208
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8254..8682
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8341..8360)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8551..8572
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8691..8711)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8696..9025
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8876..9011
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8938..8957
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    9848..9969
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10018..10034)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10018..10034)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10031..10053)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10042..10058
                     /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(10066..10072,10073..10090,10091..10096))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10111..10132
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10249..10266)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10420..11008)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10500..10519)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11179..11970,11971..12039))
                     /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     11802..11821
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12040..12144)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    12192..12326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             12494..13127
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    13174..13299
                     /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 gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc
     2281 tatcagtgat agagaaaagt gaaagtcgag ctcggtaccc gggtcgaggt aggcgtgtac
     2341 ggtgggaggc ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc
     2401 atccacgctg ttttgacctc catagaagac accgggaccg atccagccta ccggtatgga
     2461 gcagaagctg atctccgagg aggacctggg atctgggcat atggcggcgg cggcggcggc
     2521 gggcgcgggc ccggagatgg tccgcgggca ggtgttcgac gtggggccgc gctacaccaa
     2581 cctctcgtac atcggcgagg gcgcctacgg catggtgtgc tctgcttatg ataatgtcaa
     2641 caaagttcga gtagctatca agaaaatcag cccctttgag caccagacct actgccagag
     2701 taccctgagg gagataaaaa tcttactgcg cttcagacat gagaacatca ttggaatcaa
     2761 tgacattatt cgagcaccaa ccatcgagca aatgaaagat gtatatatag tacaggacct
     2821 catggaaaca gatctttaca agctcttgaa gacacaacac ctcagcaatg accatatctg
     2881 ctattttctc taccagatcc tcagagggtt aaaatatatc cattcagcta acgttctgca
     2941 ccgtgacctc aagcctgaca acctgctgct caacaccacc tgtgatctca agatctgtga
     3001 ctttggcctg gcccgtgttg cagatccaga ccatgatcac acagggttcc tgacagaata
     3061 tgtggccaca cgttggtaca gggctccaga aattatgttg aattccaagg gctacaccaa
     3121 gtccattgat atttggtctg taggctgcat tctggcagaa atgctttcta acaggcccat
     3181 ctttccaggg aagcattatc ttgaccagct gaaccacatt ttgggtattc ttggatcccc
     3241 atcacaagaa gacctgaatt gtataataaa tttaaaagct aggaactatt tgctttctct
     3301 tccacacaaa aataaggtgc catggaacag gctgttccca aatgctgact ccaaagctct
     3361 ggacttattg gacaaaatgt tgacattcaa cccacacaag aggattgaag tagaacaggc
     3421 tctggcccac ccatatctgg agcagtatta cgacccgagt aacgagccca tcgccgaagc
     3481 accattcaag ttcgacatgg aattggatga cttgcctaag gaaaagctca aagaactaat
     3541 ttttgaagag actgctagat tccagccagg atacagatct taaacgcgta agatctggcc
     3601 tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc ctcacggcga gcgctgccac
     3661 gtcagacgaa gggcgcagcg agcgtcctga tccttccgcc cggacgctca ggacagcggc
     3721 ccgctgctca taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg
     3781 ggacttgggt gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa
     3841 gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt
     3901 atataaggac gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg
     3961 gttcttgttt gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc
     4021 ggggctttcg tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag
     4081 ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg ggggttgggg ggagcgcagc
     4141 aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc
     4201 gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc aaggtcttga ggccttcgct
     4261 aatgcgggaa agctcttatt cgggtgagat gggctggggc accatctggg gaccctgacg
     4321 tgaagtttgt cactgactgg agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg
     4381 gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga
     4441 cgtcacccgt tctgttggct tataatgcag ggtggggcca cctgccggta ggtgtgtggt
     4501 aggcttttct ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc ctgaatcgac
     4561 aggcgccgga cctctggtga ggggagggat aagtgaggcg tcagtttctt tggtcggttt
     4621 tatgtaccta tcttcttaag tagctgaagc tccggttttg aactatgcgc tcggggttgg
     4681 cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt gggtcaatat
     4741 gtaattttca gtgttagact agtaaattgt ccgctaaatt ctggccgttt ttggcttttt
     4801 tgttagacgg atccataact tcgtataatg tatgctatac gaagttatgc caccatgtct
     4861 agactggaca agagcaaagt cataaacgga gctctggaat tactcaatgg tgtcggtatc
     4921 gaaggcctga cgacaaggaa actcgctcaa aagctgggag ttgagcagcc taccctgtac
     4981 tggcacgtga agaacaagcg ggccctgctc gatgccctgc caatcgagat gctggacagg
     5041 catcataccc acttctgccc cctggaaggc gagtcatggc aagactttct gcggaacaac
     5101 gccaagtcat accgctgtgc tctcctctca catcgcgacg gggctaaagt gcatctcggc
     5161 acccgcccaa cagagaaaca gtacgaaacc ctggaaaatc agctcgcgtt cctgtgtcag
     5221 caaggcttct ccctggagaa cgcactgtac gctctgtccg ccgtgggcca ctttacactg
     5281 ggctgcgtat tggaggaaca ggagcatcaa gtagcaaaag aggaaagaga gacacctacc
     5341 accgattcta tgcccccact tctgagacaa gcaattgagc tgttcgaccg gcagggagcc
     5401 gaacctgcct tccttttcgg cctggaacta atcatatgtg gcctggagaa acagctaaag
     5461 tgcgaaagcg gcgggccgac cgacgccctt gacgattttg acttagacat gctcccagcc
     5521 gatgcccttg acgactttga ccttgatatg ctgcctgctg acgctcttga cgattttgac
     5581 cttgacatgc tccccgggta aataacttcg tatagcatac attatacgaa gtatggcgcg
     5641 ccaattccgc ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata
     5701 aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt
     5761 gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct
     5821 cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc
     5881 ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga
     5941 caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc
     6001 ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt
     6061 attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg
     6121 gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc
     6181 gaaccacggg gacgtggttt tcctttgaaa aacacgataa taccatggcc accgagtaca
     6241 agcccacggt gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc accctcgccg
     6301 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga cccggaccgc cacatcgagc
     6361 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     6421 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     6481 gggcggtgtt cgccgagatc ggctcgcgca tggccgagtt gagcggttcc cggctggccg
     6541 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     6601 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     6661 tccccggagt ggaggcggcc gagcgcgctg gggtgcccgc cttcctggag acctccgcgc
     6721 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     6781 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgatta attaagcgcg
     6841 tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     6901 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     6961 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     7021 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     7081 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     7141 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     7201 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat
     7261 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     7321 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     7381 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     7441 gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta gcaatacagc
     7501 agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc
     7561 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     7621 ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa gacaagatct
     7681 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     7741 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     7801 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     7861 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     7921 aaagaaatga atatcagaga gtgagaggcc ttgacattgt ttaaacccgc tgatcagcct
     7981 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc cttccccgtg ccttccttga
     8041 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     8101 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     8161 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     8221 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     8281 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     8341 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     8401 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     8461 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     8521 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     8581 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     8641 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     8701 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     8761 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     8821 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     8881 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     8941 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     9001 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     9061 tgatcagcac gtgatgaaaa agcctgaact caccgcgacg tgaccgcaag gaatcggtca
     9121 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     9181 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     9241 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     9301 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     9361 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     9421 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg
     9481 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     9541 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     9601 tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga
     9661 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca
     9721 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     9781 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     9841 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     9901 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactaatcaa
     9961 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
    10021 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
    10081 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
    10141 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
    10201 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
    10261 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
    10321 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
    10381 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
    10441 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
    10501 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
    10561 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
    10621 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacta
    10681 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
    10741 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
    10801 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
    10861 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
    10921 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
    10981 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
    11041 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
    11101 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
    11161 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
    11221 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
    11281 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
    11341 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
    11401 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
    11461 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
    11521 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
    11581 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
    11641 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
    11701 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
    11761 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
    11821 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
    11881 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
    11941 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
    12001 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
    12061 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
    12121 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
    12181 atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
    12241 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
    12301 tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa aatcatccca
    12361 aacttcccac cccataccct attaccactg ccaattacct gtggtttcat ttactctaaa
    12421 cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa atatgtacta
    12481 caaacttagt agttggaagg gctaattcac tcccaaagaa gacaagatat ccttgatctg
    12541 tggatctacc acacacaagg ctacttccct gattagcaga actacacacc agggccaggg
    12601 gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga gccagataag
    12661 gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag cctgcatggg
    12721 atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct agcatttcat
    12781 cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg agcttgctac
    12841 aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac tggggagtgg
    12901 cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt ctctctggtt
    12961 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
    13021 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
    13081 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac
    13141 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct
    13201 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact
    13261 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaag
//
