LOCUS       sequence-330703-       10637 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 10637)
  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 10637)
  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_EKAREN4-ires-blast
FEATURES             Location/Qualifiers
     source          1..10637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    73..625
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(217..234)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     444..463
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      646..655
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             652..1050
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             1899..2132
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2204..2325
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2241..2260)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2295..2314
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2365..2500
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2520..2539)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2521..2539)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2549..2566)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2549..2565)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2558..2580)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2707..3162
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2794..2813)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3004..3025
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3188..3292
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3293..3361,3362..4153)
                     /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(3511..3530)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4324..4912
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4813..4832
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5066..5083
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5200..5221
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5236..5241,5242..5259,5260..5266)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5274..5290
                     /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     5279..5301
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5298..5314
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5298..5314
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5333..5353
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5335..5353
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5381..5607
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5608..5788
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5835..5960
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6453..6686
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6871..6915
                     /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    7177..7294
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7322..8484
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          7553..8475
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     8432..8452
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(8566..8568,8569..8571,8572..9279)
                     /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(8872..8891)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             9454..9801
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             10588..10608
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 cggtcgttaa cattaagcta gcctagtgcc atttgttcag tggcccggga attccgcccc
       61 tctccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
      121 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
      181 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
      241 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
      301 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
      361 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
      421 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
      481 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
      541 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
      601 tcctttgaaa aacacgatga taatatggat atggccacaa cggtcgccac catggccaag
      661 cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc
      721 cccatctctg aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc
      781 actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc
      841 actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac
      901 aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg
      961 atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg
     1021 ctgccctctg gttatgtgtg ggagggctaa tacgtaatgc atgtattcaa tctaagcagg
     1081 ctttcacttt ctcgccaact tacaaggcct ttctgtgtaa acaatacctg aacctttacc
     1141 ccgttgcccg gcaacggcca ggtctgtgcc aagtgtttgc tgacgcaacc cccactggct
     1201 ggggcttggt catgggccat cagcgcatgc gtggaacctt ttcggctcct ctgccgatcc
     1261 atactgcgga actcctagcc gcttgttttg ctcgcagcag gtctggagca aacattatcg
     1321 ggactgataa ctctgttgtc ctatcccgca aatatacatc gtttccatgg ctgctaggct
     1381 gtgctgccaa ctggatcctg cgcgggacgt cctttgttta cgtcccgtcg gcgctgaatc
     1441 ctgcggacga cccttctcgg ggtcgcttgg gactctctcg tccccttctc cgtctgccgt
     1501 tccgaccgac cacggggcgc acctctcttt acgcggactc cccgtctgtg ccttctcatc
     1561 tgccggaccg tgtgcacttc gcttcacctc tgcacgtcgc atggagacca ccgtgaacgc
     1621 ccaccaaata ttgcccaagg tcttacataa gaggactctt ggactctcag caatgtcaac
     1681 gaccgacctt gaggcatact tcaaagactg tttgtttaaa gactgggagg agttggggga
     1741 ggagattagg ttaaaggtct ttgtactagg aggctgtagg cataaattgg tctgcgcacc
     1801 agcaccatgg cgcaatcact agagcggggt acctttaaga ccaatgactt acaaggcagc
     1861 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     1921 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     1981 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     2041 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     2101 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     2161 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
     2221 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2281 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
     2341 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2401 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     2461 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc
     2521 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
     2581 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     2641 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     2701 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2761 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2821 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2881 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     2941 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3001 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3061 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3121 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     3181 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3241 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3301 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3361 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3421 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3481 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3541 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3601 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3661 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3721 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3781 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3841 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3901 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3961 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4021 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4081 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4141 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4201 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4261 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4321 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4381 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4441 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4501 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4561 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4621 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4681 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4741 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4801 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4861 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4921 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4981 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     5041 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     5101 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     5161 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     5221 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     5281 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
     5341 ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc
     5401 ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag
     5461 caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa
     5521 cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta
     5581 tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg
     5641 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     5701 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     5761 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     5821 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     5881 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     5941 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     6001 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     6061 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     6121 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     6181 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     6241 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     6301 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     6361 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     6421 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     6481 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
     6541 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     6601 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     6661 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     6721 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
     6781 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
     6841 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
     6901 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
     6961 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
     7021 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
     7081 aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga
     7141 tccattcgat tagtgaacgg atctcgacgg tatcggtttt aaaagaaaag gggggattgg
     7201 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
     7261 attacaaaaa caaattacaa aaattcaaaa ttttatcgat catatgacta gagcggcctt
     7321 aggctccggt gcgcgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
     7381 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
     7441 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     7501 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
     7561 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
     7621 tacttccacc tggctccagt acgtgattct tgatcccgag ctggagccag gggcgggcct
     7681 tgcgctttag gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc
     7741 cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta
     7801 gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta
     7861 aatgcgggcc aggatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg
     7921 gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga
     7981 atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg
     8041 tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa
     8101 agatggccgc ttcccggccc tgctccaggg ggctcaaaat ggaggacgcg gcgctcggga
     8161 gagcgggcgg gtgagtcacc cacacaaagg aaaggggcct ttccgtcctc agccgtcgct
     8221 tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctggagcttt
     8281 tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact
     8341 gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt
     8401 gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt
     8461 tttcttccat ttcaggtgtc gtgattaatt aaacgcgtac cgatagcttc gaattctgca
     8521 gtcgacgata tcactagtct cgagcatttt ggcaaagaat tcggcatggt gagcaagggc
     8581 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     8641 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     8701 aagcttctat gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     8761 ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     8821 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     8881 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     8941 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     9001 tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     9061 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     9121 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     9181 tccgccctgt tcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcctg
     9241 accgccgccg ggatcactga gggcatgaac gagctgtacc tcgagatggc ggacgaggag
     9301 aagctgccgc ccggctggga gaagcgcatg agccgcagct caggccgagt gtactacttc
     9361 aaccacatca ctaacgccag ccagtgggag cggcccagcg gcaacagcag cagtggtggc
     9421 aaaaacgggc agggggagcc tgccaggggt accagtgctg gtggtagtgc tggtggtagt
     9481 gctggtggta gtgctggtgg tagtgctggt ggttccggca gtgctggtgg tagtgctggt
     9541 ggtagtacca gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc tggtggtagt
     9601 gctggtggtt ccggcagtgc tggtggtagt gctggtggta gtaccagtgc tggtggtagt
     9661 gctggtggta gtgctggtgg tagtgctggt ggtagtgctg gtggttccgg cagtgctggt
     9721 ggtagtgctg gtggtagtac cagtgctggt ggtagtgctg gtggtagtgc tggtggtagt
     9781 gctggtggta gtgctggtgg ttccggacca gatgtcccta gaactccagt ggatccagca
     9841 tggctgtcat tccaatttcc gggcggccgc atggtgagta aaggggaaga actgtttact
     9901 ggcgtggtgc caattctggt cgaactggat ggagatgtaa atgggcataa atttagtgtg
     9961 tctggagaag gggaaggaga cgctacatat gggaaactga cactgaaatt tatttgtact
    10021 actggaaaac tgcctgtgcc ttggcctaca ctcgtgacaa ccctgtcttg gggagtgcag
    10081 tgttttgccc ggtatcctga tcatatgaaa cagcatgatt ttttcaaatc tgccatgcct
    10141 gagggatatg tccaggaacg gacaattttc tttaaagatg atgggaatta taaaacacgg
    10201 gccgaagtga aatttgaagg ggatacactg gtgaatcgga ttgaactgaa aggaattgat
    10261 tttaaagaag atgggaatat tctgggccat aaactggaat ataattattt cagcgataat
    10321 gtctacatta cagccgataa acagaagaat gggattaaag ccaattttaa aattcggcat
    10381 aatattgaag atggaggggt gcagctcgcc gatcattatc agcagaatac acctattggg
    10441 gatgggcctg tgctgctgcc tgataatcat tatctgagca cacagtctgc cctgagcaag
    10501 gaccctaatg aaaagcggga ccatatggtc ctgctggaat ttctgactgc cgccggcatt
    10561 acactcggga tggatgaact gtctagacct aagaagaagc gtaaggtgta agtcgacggt
    10621 cgagtcgaga agctagc
//
