LOCUS       sequence-260250-        9414 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Visualization of Akt and ERK activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9414)
  AUTHORS   Chavez-Abiega S, Gronloh MLB, Gadella TWJ, Bruggeman FJ, Goedhart J
  TITLE     Single-cell imaging of ERK and Akt activation dynamics and 
            heterogeneity induced by G-protein-coupled receptors.
  JOURNAL   J Cell Sci. 2022 Mar 15;135(6):jcs259685. doi: 10.1242/jcs.259685. 
            Epub 2022 Mar 11.
  PUBMED    35107584
REFERENCE   2  (bases 1 to 9414)
  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: "J Cell 
            Sci."; date: "2022-03-15"; volume: "135(6):jcs259685. doi"; pages: "
            10.1242/jcs.259685. Epub 2022 Mar 11"
COMMENT     Sequence Label: PB-H2A-mScarlet-2A-AKT_KTR-mTq2-2A-ERK_KTR-mNG
FEATURES             Location/Qualifiers
     source          1..9414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   13..47
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        440..975
                     /label=mCMV promoter
                     /note="mouse cytomegalovirus (CMV) immediate early 
                     promoter"
     intron          999..1941
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1898..1918
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      2399..2408
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2405..3097
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELY"
     primer_bind     complement(2538..2556)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2806..2825
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3116..3172
                     /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"
     regulatory      4379..4388
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4385..4387,4388..4390,4391..5098)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(4430..4451)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4691..4710)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5038..5059
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5117..5173
                     /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"
     regulatory      5435..5444
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5441..6151
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      5696..5705
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6179..6197)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6180..6197)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6217..6234)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6223..6447
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(6579..7167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6659..6678)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    7242..7376
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7279..7298)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7333..7352
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(7381..7980)
                     /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(7464..7484)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7961..7980
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     repeat_region   8383..8445
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     CDS             complement(join(8537..9328,9329..9397))
                     /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     9160..9179
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggcgcgcctt aaccctagaa agatagtctg cgtaaaattg acgcatgcat tcttgaaata
       61 ttgctctctc tttctaaata gcgcgaatcc gtcgctgtgc atttaggaca tctcagtcgc
      121 cgcttggagc tcccgtgagg cgtgcttgtc aatgcggtaa gtgtcactga ttttgaacta
      181 taacgaccgc gtgagtcaaa atgacgcatg attatctttt acgtgacttt taagatttaa
      241 ctcatacgat aattatattg ttatttcatg ttctacttac gtgataactt attatatata
      301 tattttcttg ttatagatat catcgataac aggaaagttc cattggagcc aagtacattg
      361 agtcaatagg gactttccaa tgggttttgc ccagtacata aggtcaatgg gaggtaagcc
      421 aatgggtttt tcccattact ggcacgtata ctgagtcatt agggactttc caatgggttt
      481 tgcccagtac ataaggtcaa taggggtgaa tcaacaggaa agttccattg gagccaagta
      541 cactgagtca atagggactt tccattgggt tttgcccagt acaaaaggtc aatagggggt
      601 gagtcaatgg gtttttccca ttattggcac gtacataagg tcaatagggg tgagtcattg
      661 ggtttttcca gccaatttaa ttaaaacgcc atgtactttc ccaccattga cgtcaatggg
      721 ctattgaaac taatgcaacg tgacctttaa acggtacttt cccatagctg attaatggga
      781 aagtaccgtt ctcgagccaa tacacgtcaa tgggaagtga aagggcagcc aaaacgtaac
      841 accgccccgg ttttcccctg gaaattccat attggcacgc attctattgg ctgagctgcg
      901 ttctacgtgg gtataagagg cgcgaccagc gtcggtaccg tcgcagtctt cggtctgacc
      961 accgtagaac gcagagctcc tcgctgcagg caagcttggt aagtgccgtg tgtggttccc
     1021 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacgcccc
     1081 tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg
     1141 aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc
     1201 tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag
     1261 tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt
     1321 cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc
     1381 gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca
     1441 ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg
     1501 ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga
     1561 gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc
     1621 tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc
     1681 gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg
     1741 agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc
     1801 cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg
     1861 gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa
     1921 agtttttttc ttccatttca gggatccact agcgctaccg gactcagatc tcgagctcaa
     1981 gcttcgaatt ctgatgtcgg gacgcggcaa gcagggaggc aaagctcgcg ccaaagccaa
     2041 gacccgctct tctcgtgccg gtctccagtt ccccgtgggc cgagtgcacc gactgctccg
     2101 caagggcaac tatgctgagc gggtcggggc cggcgcgccg gtgtacctgg cggcggtgct
     2161 ggagtacctg actgccgaga tcctggagct ggcgggcaac gccgcccgcg acaacaagaa
     2221 gacccgcatt atcccgcgcc acttgcagct ggccatccgc aacgacgagg agctcaacaa
     2281 gctgctgggc aaagtaacca tcgctcaggg tggtgtcctg cccaacatcc aggctgtgct
     2341 actgcccaag aagaccgaga gtcaccacaa ggccaaaggc aaacgggatc caccggtcgc
     2401 caccatggtg agcaagggcg aggcagtgat caaggagttc atgcggttca aggtgcacat
     2461 ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     2521 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttctcctg
     2581 ggacatcctg tcccctcagt tcatgtacgg ctccagggcc ttcatcaagc accccgccga
     2641 catccccgac tactataagc agtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     2701 cttcgaggac ggcggcgccg tgaccgtgac ccaggacacc tccctggagg acggcaccct
     2761 gatctacaag gtgaagctcc gcggcaccaa cttccctcct gacggccccg taatgcagaa
     2821 gaagacaatg ggctgggaag cgtccaccga gcggttgtac cccgaggacg gcgtgctgaa
     2881 gggcgacatt aagatggccc tgcgcctgaa ggacggcggc cgctacctgg cggacttcaa
     2941 gaccacctac aaggccaaga agcccgtgca gatgcccggc gcctacaacg tcgaccgcaa
     3001 gttggacatc acctcccaca acgaggacta caccgtggtg gaacagtacg aacgctccga
     3061 gggccgccac tccaccggcg gcatggacga gctgtaccaa accatggggt caggggccac
     3121 caacttctct cttctgaaac aagccggcga cgtcgaggaa aaccccggac ctatggcaga
     3181 ggcaccggct tccccggccc cgctctctcc gctcgaagtg gagctggacc cggagttcga
     3241 gccccagagc cgtccgcgat cctgtacgtg gcccctgcaa aggccggagc tccaagcgag
     3301 ccctgccaag ccctcggggg agacggccgc cgactccatg atccccgagg aggaggacga
     3361 tgaagacgac gaggacggcg ggggacgggc cggctcggcc atggcgatcg gcggcggcgg
     3421 cgggagcggc acgctgggct ccgggctgct ccttgaggac tcggcccggg tgctggcacc
     3481 cggagggcaa gaccccgggt ctgggccagc caccgcggcg ggcgggctga gcgggggtac
     3541 acaggcgctg ctgcagcctc agcaaccgct gccaccgccg cagccggggg tggctggggg
     3601 ctccgggcag ccgaggaaat gttcgtcgcg gcggaacgcc tggggaaacc tgtcctacgc
     3661 ggacctgatc acccgcgcca tcgagagctc cccggacaaa cggctcactc tgtcccagat
     3721 ctacgagtgg atggtgcgtt gcgtgcccta cttcaaggat aagggcgaca gcaacagctc
     3781 tgccggctgg aagaactcca tccggcgcaa cctgtcactg catagtcgat tcatgcgggt
     3841 ccagaatgag ggaactggca agagctcttg gtggatcatc aaccctgatg gggggaagag
     3901 cggaaaagcc ccccggcggc gggctgtctc catggacaat agcaacaagt ataccaagag
     3961 ccgtggccgc gcagccaaga agaaggcagc cctgcagaca gcccccgaat cagctgacga
     4021 cagtccctcc cagctctcca agtggcctgg cgcccccacg tcacgcagca gtgatgagct
     4081 ggatgcgtgg acggacttcc gttcacgcac caattctaac gccagcacag tcagtggccg
     4141 cctgtcgccc atcatggcaa gcacagagtt ggatgaagtc caggacgatg atgcgcctct
     4201 cgcgcccatg ctctacagca gctcagccag cctgtcacct tcagtaagca agccgtgcac
     4261 ggtggaactg ccacggctga ctgatatggc aggcaccatg aatctgaatg atgggctgac
     4321 tgaaaacctc atggacgacc tgctggataa catcacgctc ggcggccgca taccggtcgc
     4381 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     4441 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     4501 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     4561 caccctcgtg accaccctgt cctggggcgt gcagtgcttc gcccgctacc ccgaccacat
     4621 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     4681 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     4741 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     4801 gcacaagctg gagtacaact actttagcga caacgtctat atcaccgccg acaagcagaa
     4861 gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct
     4921 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     4981 ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     5041 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacca
     5101 aaccatgggg agcggggcta ccaacttctc cctcctcaag caggcagggg acgtcgagga
     5161 aaatcctgga cccatgaaag gccgcaagcc tcgcgatctg gagttgcctc tcagccctag
     5221 cctcctgggg ggacagggcc ctgagcgcac acccgggtcc ggcacctcta gtggtttgca
     5281 ggctccggga cctgccctgt caccctccaa gcgcagcggc ctggaggatc ctgccacacc
     5341 cagcaaaaaa ccaaggaccc cctcagtcag ctcgcggtta gagcgactga cgctccagtc
     5401 ttctttccaa ttcccgtccg gcggcatacc ggtcgccacc atggtgagca agggcgagga
     5461 ggataacatg gcctctctcc cagcgacaca tgagttacac atctttggct ccatcaacgg
     5521 tgtggacttt gacatggtgg gtcagggcac cggcaatcca aatgatggtt atgaggagtt
     5581 aaacctgaag tccaccaagg gtgacctcca gttctccccc tggattctgg tccctcatat
     5641 cgggtatggc ttccatcagt acctgcccta ccctgacggg atgtcgcctt tccaggccgc
     5701 catggtagat ggctccggct accaagtcca tcgcacaatg cagtttgaag atggtgcctc
     5761 ccttactgtt aactaccgct acacctacga gggaagccac atcaaaggag aggcccaggt
     5821 gaaggggact ggtttccctg ctgacggtcc tgtgatgacc aactcgctga ccgctgcgga
     5881 ctggtgcagg tcgaagaaga cttaccccaa cgacaaaacc atcatcagta cctttaagtg
     5941 gagttacacc actggaaatg gcaagcgcta ccggagcact gcgcggacca cctacacctt
     6001 tgccaagcca atggcggcta actatctgaa gaaccagccg atgtacgtgt tccgtaagac
     6061 ggagctcaag cactccaaga ccgagctcaa cttcaaggag tggcaaaagg cctttaccga
     6121 tgtgatgggc atggacgagc tgtacaagta aagcggccgc gactctagag ggccctattc
     6181 tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     6241 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     6301 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     6361 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     6421 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagt ggcggtaata
     6481 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6541 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6601 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6661 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6721 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6781 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6841 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6901 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6961 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     7021 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     7081 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     7141 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     7201 gctcagtgga acgaaaactc acgttaaggg attttggtca taacttgttt attgcagctt
     7261 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     7321 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct
     7381 tcaggcaccg ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc
     7441 ggcggtgacg gtgaagccga gccgctcgta gaaggggagg ttgcggggcg cggaggtctc
     7501 caggaaggcg ggcaccccgg cgcgctcggc cgcctccact ccggggagca cgacggcgct
     7561 gcccagaccc ttgccctggt ggtcgggcga gacgccgacg gtggccagga accacgcggg
     7621 ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt tgctgcgcgg ccagcctgga
     7681 accgctcaac tcggccatgc gcgggccgat ctcggcgaac accgcccccg cttcgacgct
     7741 ctccggcgtg gtccagaccg ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat
     7801 gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg
     7861 gtccgggtcg acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg
     7921 tacggcccgg gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat
     7981 ggtggcctgc agagtcgctc tgtgttcgag gccacacgcg tcaccttaat atgcgaagtg
     8041 gacctgggac cgcgccgccc cgactgcatc tgcgtgtttt cgccaatgac aagacgctgg
     8101 gcggggtttg tgtcatcata gaactaaaga catgcaaata tatttcttcc ggggacaccg
     8161 ccagcaaacg cgagcaacgg gccacgggga tgaagcagct ggctagctaa aagttttgtt
     8221 actttataga agaaattttg agtttttgtt tttttttaat aaataaataa acataaataa
     8281 attgtttgtt gaatttatta ttagtatgta agtgtaaata taataaaact taatatctat
     8341 tcaaattaat aaataaacct cgatatacag accgataaaa cacatgcgtc aattttacgc
     8401 atgattatct ttaacgtacg tcacaatatg attatctttc tagggttaat tcgaacagct
     8461 ggttctttcc gcctcaggac tcttcctttt tcaataaatc aatctaaagt atatatgagt
     8521 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     8581 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     8641 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     8701 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     8761 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     8821 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     8881 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     8941 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     9001 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     9061 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     9121 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     9181 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     9241 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     9301 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     9361 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caat
//
