LOCUS       sequence-338960-       10487 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optimized EKAREV FRET biosensor sensor for ERK.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10487)
  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 10487)
  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: pLV x EF1a_EKAREN5-ires-blast
FEATURES             Location/Qualifiers
     source          1..10487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    51..603
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(195..212)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     422..441
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      624..633
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             630..1028
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             1871..2104
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2176..2297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2213..2232)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2267..2286
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2337..2472
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2492..2511)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2493..2511)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2521..2538)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2521..2537)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2530..2552)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2679..3134
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2766..2785)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2976..2997
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3160..3264
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3265..3333,3334..4125)
                     /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     complement(3483..3502)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4296..4884
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4785..4804
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5038..5055
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5172..5193
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5208..5213,5214..5231,5232..5238)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5246..5262
                     /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     5251..5273
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5270..5286
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5270..5286
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5305..5325
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5307..5325
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5353..5579
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5580..5760
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5807..5932
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6425..6658
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6843..6887
                     /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    7149..7266
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7283..8445
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          7514..8436
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     8393..8413
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(8536..8538,8539..8541,8542..9249)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLLCTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLL
                     EFLTAAGITEGMNELY"
     primer_bind     complement(8842..8861)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             10438..10458
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ctagtgccat ttgttcagtg gcccgggaat tccgcccctc tcccccccta acgttactgg
       61 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt
      121 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
      181 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
      241 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
      301 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
      361 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
      421 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
      481 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa
      541 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata
      601 atatggatat ggccacaacg gtcgccacca tggccaagcc tttgtctcaa gaagaatcca
      661 ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg
      721 tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt
      781 ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg
      841 gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg
      901 gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca
      961 gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg
     1021 agggctaaat gcatgtattc aatctaagca ggctttcact ttctcgccaa cttacaaggc
     1081 ctttctgtgt aaacaatacc tgaaccttta ccccgttgcc cggcaacggc caggtctgtg
     1141 ccaagtgttt gctgacgcaa cccccactgg ctggggcttg gtcatgggcc atcagcgcat
     1201 gcgtggaacc ttttcggctc ctctgccgat ccatactgcg gaactcctag ccgcttgttt
     1261 tgctcgcagc aggtctggag caaacattat cgggactgat aactctgttg tcctatcccg
     1321 caaatataca tcgtttccat ggctgctagg ctgtgctgcc aactggatcc tgcgcgggac
     1381 gtcctttgtt tacgtcccgt cggcgctgaa tcctgcggac gacccttctc ggggtcgctt
     1441 gggactctct cgtccccttc tccgtctgcc gttccgaccg accacggggc gcacctctct
     1501 ttacgcggac tccccgtctg tgccttctca tctgccggac cgtgtgcact tcgcttcacc
     1561 tctgcacgtc gcatggagac caccgtgaac gcccaccaaa tattgcccaa ggtcttacat
     1621 aagaggactc ttggactctc agcaatgtca acgaccgacc ttgaggcata cttcaaagac
     1681 tgtttgttta aagactggga ggagttgggg gaggagatta ggttaaaggt ctttgtacta
     1741 ggaggctgta ggcataaatt ggtctgcgca ccagcaccat ggcgcaatca ctagagcggg
     1801 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     1861 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     1921 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     1981 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     2041 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     2101 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     2161 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2221 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     2281 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc
     2341 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     2401 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     2461 tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc
     2521 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     2581 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     2641 cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt
     2701 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2761 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2821 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2881 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2941 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3001 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3061 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3121 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3181 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3241 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3301 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3361 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3421 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     3481 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     3541 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3601 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3661 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3721 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3781 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     3841 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3901 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     3961 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     4021 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4081 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4141 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4201 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4261 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4321 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4381 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4441 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4501 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4561 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4621 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4681 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4741 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4801 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4861 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4921 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4981 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     5041 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     5101 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     5161 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     5221 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     5281 tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag
     5341 ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga
     5401 gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa
     5461 ggtggtacga tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa
     5521 ccactgaatt gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg
     5581 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     5641 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     5701 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     5761 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
     5821 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
     5881 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
     5941 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
     6001 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
     6061 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
     6121 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
     6181 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
     6241 aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag
     6301 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     6361 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     6421 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     6481 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     6541 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     6601 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     6661 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     6721 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     6781 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     6841 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     6901 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     6961 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     7021 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     7081 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac
     7141 ggtatcggtt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     7201 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     7261 aattttatcg ataagcttgg gtggctccgg tgcccgtcag tgggcagagc gcacatcgcc
     7321 cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg
     7381 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     7441 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     7501 cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta
     7561 tggcccttgc gtgccttgaa ttacttccac ctggctccag tacgtgattc ttgatcccga
     7621 gctggagcca ggggcgggcc ttgcgcttta ggagcccctt cgcctcgtgc ttgagttgag
     7681 gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc
     7741 gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt
     7801 ttctggcaag atagtcttgt aaatgcgggc caggatctgc acactggtat ttcggttttt
     7861 ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc
     7921 ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg
     7981 gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg
     8041 gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctccagg gggctcaaaa
     8101 tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaggggcc
     8161 tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac
     8221 ctcgattagt tctggagctt ttggagtacg tcgtctttag gttgggggga ggggttttat
     8281 gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg
     8341 atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct
     8401 cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaggatc cccgggcgcg
     8461 ccacgcgtac cgatagcttc gaattctgca gtcgacgata tcactagtct cgagcatttt
     8521 ggcaaagaat tcggcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     8581 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     8641 ggcgatgcca cctacggcaa gctgaccctg aagcttctat gcaccaccgg caagctgccc
     8701 gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt cgcccgctac
     8761 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     8821 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     8881 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     8941 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc
     9001 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     9061 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     9121 ctgcccgaca accactacct gagctaccag tccgccctgt tcaaagaccc caacgagaag
     9181 cgcgatcaca tggtcctgct ggagttcctg accgccgccg ggatcactga gggcatgaac
     9241 gagctgtacc tcgagatggc ggacgaggag aagctgccgc ccggctggga gaagcgcatg
     9301 agccgcagct caggccgagt gtactacttc aaccacatca ctaacgccag ccagtgggag
     9361 cggcccagcg gcaacagcag cagtggtggc aaaaacgggc agggggagcc tgccaggggt
     9421 accagtgctg gtggtagtgc tggtggtagt gctggtggtt ccggcagtgc tggtggtagt
     9481 gctggtggta gtaccagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt
     9541 ggtagtgctg gtggttccgg cagtgctggt ggtagtgctg gtggtagtac cagtgctggt
     9601 ggtagtgctg gtggtagtgc tggtggtagt gctggtggta gtgctggtgg ttccggacca
     9661 gatgtcccta gaactccagt ggatccagca tggctgtcat tccaatttcc gggcggccgc
     9721 atggtgagta aaggggaaga actgtttact ggcgtggtgc caattctggt cgaactggat
     9781 ggagatgtaa atgggcataa atttagtgtg tctggagaag gggaaggaga cgctacatat
     9841 gggaaactga cactgaaatt tatttgtact actggaaaac tgcctgtgcc ttggcctaca
     9901 ctcgtgacaa ccctgtcttg gggagtgcag tgttttgccc ggtatcctga tcatatgaaa
     9961 cagcatgatt ttttcaaatc tgccatgcct gagggatatg tccaggaacg gacaattttc
    10021 tttaaagatg atgggaatta taaaacacgg gccgaagtga aatttgaagg ggatacactg
    10081 gtgaatcgga ttgaactgaa aggaattgat tttaaagaag atgggaatat tctgggccat
    10141 aaactggaat ataattattt cagcgataat gtctacatta cagccgataa acagaagaat
    10201 gggattaaag ccaattttaa aattcggcat aatattgaag atggaggggt gcagctcgcc
    10261 gatcattatc agcagaatac acctattggg gatgggcctg tgctgctgcc tgataatcat
    10321 tatctgagca cacagtctgc cctgagcaag gaccctaatg aaaagcggga ccatatggtc
    10381 ctgctggaat ttctgactgc cgccggcatt acactcggga tggatgaact gtctagacct
    10441 aagaagaagc gtaaggtgta agtcgacggt cgagtcgaga cgctagc
//
