LOCUS       sequence-330719-       11293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of dominant negative HRas(N17) and P2A-coupled 
            reporter mKate2-NLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11293)
  AUTHORS   Ponsioen B, Post JB, Buissant des Amorie JR, Laskaris D, van Ineveld
            RL, Kersten S, Bertotti A, Sassi F, Sipieter F, Cappe B, Mertens S, 
            Verlaan-Klink I, Boj SF, Vries RGJ, Rehmann H, Vandenabeele P, 
            Riquet FB, Trusolino L, Bos JL, Snippert HJG
  TITLE     Quantifying single-cell ERK dynamics in colorectal cancer organoids 
            reveals EGFR as an amplifier of oncogenic MAPK pathway signalling.
  JOURNAL   Nat Cell Biol. 2021 Apr;23(4):377-390. doi: 
            10.1038/s41556-021-00654-5. Epub 2021 Apr 1.
  PUBMED    33795873
REFERENCE   2  (bases 1 to 11293)
  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: 
            "10.1038/s41556-021-00654-5"; journalName: "Nat Cell Biol"; date: 
            "2021-04"; volume: "23"; issue: "4"; pages: "377-390"
COMMENT     Sequence Label: pInducer(p20) x mKate2-NLS-P2A-HRas(N17)_ires-...
FEATURES             Location/Qualifiers
     source          1..11293
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    join(2230..2248,2249..2271,2272..2290,2291..2313,2314..2332,
                     2333..2355,2356..2374,2375..2397,2398..2416,2417..2439,
                     2440..2458,2459..2481,2482..2500)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2533..2571
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2568..2592
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2690..3385
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     CDS             3386..3406
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3416..3436
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3443..3463
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3509..3565
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3572..4141
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=H-Ras
                     /translation="MTEYKLVVVGAGGVGKNALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVK
                     DSDDVPMVLVGNKCDLAARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREI
                     RQHKLRKLNPPDESGPGCMSCKCVLS"
     CDS             4170..4196
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        4282..5493
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5330..5349
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(5552..6169,6170..6175,6176..6211,6212..6214,6215..6250,
                     6251..6253,6254..6289,6290..6298)
                     /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    6331..6883
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6475..6492)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6702..6721
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6910..7509
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6910..6929)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7406..7426
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7683..8271
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7736..7756)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8154..8165)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8346..8579
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8698..8716)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8784..8888
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8889..8957,8958..9749)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(9107..9126)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9920..10508
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10409..10428
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10662..10679
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10919..10940
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10955..10960,10961..10978,10979..10985)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10993..11009
                     /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)."
     primer_bind     10998..11020
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11017..11033
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11017..11033
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11083..11102
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactaggatc
     2221 ctttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca
     2281 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa
     2341 agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
     2401 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
     2461 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagctcgg
     2521 tacccgggtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg
     2581 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
     2641 gaccgatcca gcctccgcgg ccccgaacta gtccagacgc gtttccacca tggtgagcga
     2701 gctgattaag gagaacatgc acatgaagct gtacatggag ggcaccgtga acaaccacca
     2761 cttcaagtgc acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgagaat
     2821 caaggcggtc gagggcggcc ctctcccctt cgccttcgac atcctggcta ccagcttcat
     2881 gtacggcagc aaaaccttca tcaaccacac ccagggcatc cccgacttct ttaagcagtc
     2941 cttccccgag ggcttcacat gggagagagt caccacatac gaagacgggg gcgtgctgac
     3001 cgctacccag gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg
     3061 ggtgaacttc ccatccaacg gccctgtgat gcagaagaaa acactcggct gggaggcctc
     3121 caccgagacc ctgtaccccg ctgacggcgg cctggaaggc agagccgaca tggccctgaa
     3181 gctcgtgggc gggggccacc tgatctgcaa cttgaagacc acatacagat ccaagaaacc
     3241 cgctaagaac ctcaagatgc ccggcgtcta ctatgtggac agaagactgg aaagaatcaa
     3301 ggaggccgac aaagagacct acgtcgagca gcacgaggtg gctgtggcca gatactgcga
     3361 cctccctagc aaactggggc acagaccaaa gaaaaagaga aaagtcggag gtggaccaaa
     3421 gaagaagcga aaagttagcg ctccaaaaaa gaaaaggaaa gtcagcatgt taacggagct
     3481 gtacaaggct agagatatcg gatctggcgc cacaaacttc tccctgttga agcaggcggg
     3541 agatgttgaa gagaaccctg gacctgctag catgacggaa tataagcttg tggtggtggg
     3601 cgccggcggt gtgggcaaga atgcgctgac catccagctg atccagaacc attttgtgga
     3661 cgaatacgac cccactatag aggattccta ccggaagcag gtggtcattg atggggagac
     3721 gtgcctgttg gacatcctgg ataccgccgg ccaggaggag tacagcgcca tgcgggacca
     3781 gtacatgcgc accggggagg gcttcctgtg tgtgtttgcc atcaacaaca ccaagtcttt
     3841 tgaggacatc caccagtaca gggagcagat caaacgggtg aaggactcgg atgacgtgcc
     3901 catggtgctg gtggggaaca agtgtgacct ggctgcacgc actgtggaat ctcggcaggc
     3961 tcaggacctc gcccgaagct acggcatccc ctacatcgag acctcggcca agacccggca
     4021 gggagtggag gatgccttct acacgttggt gcgtgagatc cggcagcaca agctgcggaa
     4081 gctgaaccct cctgatgaga gtggccccgg ctgcatgagc tgcaagtgtg tgctctcctg
     4141 actcgagtct agagggcccg cggttcgaat acccatacga cgtcccagac tacgcttagt
     4201 aatgattaat taaactagaa attctaccgg gtaggggagg cgcttttccc aaggcagtct
     4261 ggagtaaccc acccaagatc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc
     4321 ccctcctcac ggcgagcgct gccacgtcag acgaagggcg cagcgagcgt cctgatcctt
     4381 ccgcccggac gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt
     4441 atcagcagaa ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc
     4501 cagagagcgg aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc
     4561 cgtggggcgg tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc
     4621 cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg
     4681 tgagtagcgg gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac
     4741 ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg
     4801 aactgggggt tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga
     4861 cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag
     4921 aacccaaggt cttgaggcct tcgctaatgc gggaaagctc ttattcgggt gagatgggct
     4981 ggggcaccat ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc
     5041 gtctgttgcg ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg
     5101 agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg
     5161 ggccacctgc cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc
     5221 ctagggtagg ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg
     5281 aggcgtcagt ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg
     5341 ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt
     5401 gaaatgtaat catttgggtc aatatgtaat tttcagtgtt agactagtaa attgtccgct
     5461 aaattctggc cgtttttggc ttttttgtta gacgaagctt ggtaccgagc tcggatctcc
     5521 accccgtacc ggtcctgcag tcgaattcac catgtctaga ctggacaaga gcaaagtcat
     5581 aaacggagct ctggaattac tcaatggtgt cggtatcgaa ggcctgacga caaggaaact
     5641 cgctcaaaag ctgggagttg agcagcctac cctgtactgg cacgtgaaga acaagcgggc
     5701 cctgctcgat gccctgccaa tcgagatgct ggacaggcat catacccact tctgccccct
     5761 ggaaggcgag tcatggcaag actttctgcg gaacaacgcc aagtcatacc gctgtgctct
     5821 cctctcacat cgcgacgggg ctaaagtgca tctcggcacc cgcccaacag agaaacagta
     5881 cgaaaccctg gaaaatcagc tcgcgttcct gtgtcagcaa ggcttctccc tggagaacgc
     5941 actgtacgct ctgtccgccg tgggccactt tacactgggc tgcgtattgg aggaacagga
     6001 gcatcaagta gcaaaagagg aaagagagac acctaccacc gattctatgc ccccacttct
     6061 gagacaagca attgagctgt tcgaccggca gggagccgaa cctgccttcc ttttcggcct
     6121 ggaactaatc atatgtggcc tggagaaaca gctaaagtgc gaaagcggcg ggccgaccga
     6181 cgcccttgac gattttgact tagacatgct cccagccgat gcccttgacg actttgacct
     6241 tgatatgctg cctgctgacg ctcttgacga ttttgacctt gacatgctcc ccgggtaact
     6301 aagtaaggat ccgcggccgc atcgatacta acgttactgg ccgaagccgc ttggaataag
     6361 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     6421 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     6481 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     6541 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     6601 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     6661 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     6721 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     6781 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     6841 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggatat ggccacaacg
     6901 gtcgccacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     6961 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     7021 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     7081 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     7141 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     7201 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     7261 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     7321 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     7381 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     7441 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     7501 ggtgcctgac gcatgcatgc ggccgcgact ctagatcata atcagccata ccacatttgt
     7561 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     7621 gaatgcaatt gttgttgttt agtccctccc aattcgatat caagcttatc gcgatagatc
     7681 ctaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     7741 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     7801 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     7861 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     7921 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     7981 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     8041 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     8101 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     8161 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     8221 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctgagatcc
     8281 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     8341 gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg
     8401 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     8461 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     8521 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     8581 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     8641 gagtgagagg cccgggttaa ttaaggaaag ggctagatca ttcttgaaga cgaaagggcc
     8701 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     8761 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     8821 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     8881 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     8941 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     9001 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     9061 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     9121 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     9181 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     9241 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     9301 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     9361 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     9421 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     9481 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     9541 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     9601 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     9661 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     9721 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     9781 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     9841 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     9901 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     9961 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
    10021 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
    10081 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
    10141 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
    10201 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
    10261 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
    10321 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
    10381 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
    10441 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
    10501 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
    10561 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
    10621 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
    10681 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
    10741 gcagcaagct catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
    10801 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
    10861 gctccccgtg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
    10921 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
    10981 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gacatgatta
    11041 cgaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
    11101 tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa aatcatccca
    11161 aacttcccac cccataccct attaccactg ccaattacct gtggtttcat ttactctaaa
    11221 cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa atatgtacta
    11281 caaacttagt agt
//
