LOCUS       sequence-483843-       12858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express EGFP-P2A-mRFP1 in filamentous fungi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12858)
  AUTHORS   Guoliang Yuan, Tatiana Serrano-Zayas, Ziyu Dai, Erica I. DeMers, 
            Shuang Deng, Beth A. Hofstad,, Kyle R. Pomraning
  TITLE     Strong Translation Elongation Factor 1-Alpha (tef1-alpha) Promoter 
            Reporter Systems for Aspergillus niger
  JOURNAL   ACS Synthetic Biology Article ASAP
REFERENCE   2  (bases 1 to 12858)
  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: "ACS 
            Synthetic Biology Article ASAP"
COMMENT     Sequence Label: pGY232
FEATURES             Location/Qualifiers
     source          1..12858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1144..1146,1147..1857)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(1144..1146,1147..1857)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1186..1207)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1447..1466)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1794..1815
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1867..1923
                     /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             1924..2601
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1924..2601
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(2054..2072)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2322..2341
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2516..2539
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             3424..4449
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANILFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     protein_bind    complement(4521..4554)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(5024..5040)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5139..5156)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5310..5898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5390..5409)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6020..6811,6812..6880))
                     /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     6643..6662
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6881..6972)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7014..7033
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7039..7058
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    7265..7298
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     rep_origin      7611..12858
                     /label=AMA1
                     /note="replication origin from Aspergillus nidulans 
                     (Aleksenko and Clutterbuck, 1996)"
ORIGIN
        1 taacaatcta ttctccgctt ggtgatgcta aagggctttc aatagacctt gtaagtgaag
       61 gagatggagc cgtcaatccg ctaccctgcc tctggtcagt tggtctcagc aatgtaccct
      121 gtgagtcttg atagactagt tggtgactag tctctgtaga tggaacaaat ggtgcttgag
      181 agggcacgta tggcagagct ccagtcatct gcggaacata tactgggccc gggaagatct
      241 catggtcata gctgtttccg ctgagggttt aatcgttgac ctcacaggga tttccaccaa
      301 aaagtggacc gggcgggcgc cactcggccg tgccacagca gcctgagagc ggacaaataa
      361 caacagccgc ctgccgcggg gttcggttgc aaacatgacc aacaggccag gccatcatca
      421 acccaccgct gcgttgatgc ccaggatttc agtccaataa tccacaattt accaacggat
      481 agagctaggt gaactagata gacaggaggg ccagagggag gggaccgaga tgaaaaattt
      541 tcgatgaaag agtggtcaag gtggggtcgt agttcggcgc tccgagggcg aggaaccaag
      601 gaaaggcgag aaaaggacag gctgatcgcg ctgcgttgct gggctgcaag cgtgtccagt
      661 tgagtctgga aaaggctccg ccgtgaagat tctgcgttgg tcccgcacct gcgcggtggg
      721 ggcattaccc ctccatgtcc aatgatttca agtcaaagcc aagggttgaa gcccgcccgc
      781 ttagtcgcct tctcgcttga cccctccata taagtatttc ccctcctccc cctcccacaa
      841 atttttcctt tccctttcct ccctcgtccg cttcagtacg tatatcttcc ccccctctct
      901 cttccttctc actcttctct ccttctttct tgattcatcc tctctctaac tgacttcttt
      961 gctcagcacc tctacgcgtt ctggccgtag tatctgagca atttttctac agactttttc
     1021 tatctaattc caaaaaagaa cttcgagttc attcacaacc gtcatcatgg ggtgagtcaa
     1081 gacaacgatc atcatctgac aaaggaagga catgggctaa cggatatctt ctacaacagt
     1141 aaggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1201 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1261 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1321 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     1381 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1441 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1501 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1561 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     1621 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     1681 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1741 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1801 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc
     1861 tccggcgcca ccaacttctc cctcctgaag caggccggtg acgtcgagga gaaccccggc
     1921 cccatggcct cctccgagga cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag
     1981 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     2041 ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac
     2101 atcctgtccc ctcagttcca gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
     2161 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     2221 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
     2281 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     2341 accatgggct gggaggcctc caccgagcgg atgtaccccg aggacggcgc cctgaagggc
     2401 gagatcaaga tgaggctgaa gctgaaggac ggcggccact acgacgccga ggtcaagacc
     2461 acctacatgg ccaagaagcc cgtgcagctg cccggcgcct acaagaccga catcaagctg
     2521 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgagcg cgccgagggc
     2581 cgccactcca ccggcgccta agcggacatt cgatttatgc cgttatgact tccttaaaaa
     2641 agcctttacg aatgaaagaa atggaattag acttgttatg tagttgattc tacaatggat
     2701 tatgattcct gaacttcaaa tccgctgttc attattaatc tcagctcttc ccgtaaagcc
     2761 aatgttgaaa ctattcgtaa atgtacctcg ttttgcgtgt accttgctta tcacgtgata
     2821 ttacatgacc tggacagagt tctgcgcgaa agtcataacg taaatcccgg gcggtaggtg
     2881 cgtcccgggc ggaaggtagt tttctcgtcc accccaacgc gtttatcaac ctcaactttc
     2941 aacaaccatc atgccaccaa aagcgcgtaa aacaaagcga gatttgattg agcaagaggg
     3001 caggatccaa tgcgcgattc aagacattaa aaatggaaaa tttcaaaaaa ttgcgcccgc
     3061 agcgcgtgca tacaaaattc atcccaatac aagtgtaatg ctagtggagg tcaacacatc
     3121 aatgcctatt ttggtttagt cgtccaggcg gtgagcacaa aatttgtgtc gtttgacaag
     3181 atggttcatt taggcaactg gtcagatcag ccccacttgt agcagtagcg gcggcgctcg
     3241 aagtgtgact cttattagca gacaggaacg aggacattat tatcatctgc tgcttggtgc
     3301 acgataactt ggtgcgtttg tcaagcaagg taagtggacg acccggtcat accttcttaa
     3361 gttcgccctt cctcccttta tttcagattc aatctgactt acctattcta cccaagcatc
     3421 caaatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
     3481 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
     3541 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
     3601 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac
     3661 attggggagt tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg
     3721 ttgcaagacc tgcctgaaac cgaactgccc gctgttctcc agccggtcgc ggaggccatg
     3781 gatgcgattg ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa
     3841 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg
     3901 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat
     3961 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca tgcggatttc
     4021 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
     4081 gcgatgttcg gggattccca atacgaggtc gccaacatcc tcttctggag gccgtggttg
     4141 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
     4201 ccgcgcctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
     4261 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
     4321 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat
     4381 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
     4441 aaggaataga gtagatgccg accgatcccc actaccgcat taagacctca gcgcggccgc
     4501 aaatttaaat aaaatgaagt gaagttccta tactttctag agaataggaa cttctatagt
     4561 gagtcgaata agggcgacac aaaatttatt ctaaatgcat aataaatact gataacatct
     4621 tatagtttgt attatatttt gtattatcgt tgacatgtat aattttgata tcaaaaactg
     4681 attttccctt tattattttc gagatttatt ttcttaattc tctttaacaa actagaaata
     4741 ttgtatatac aaaaaatcat aaataataga tgaatagttt aattataggt gttcatcaat
     4801 cgaaaaagca acgtatctta tttaaagtgc gttgcttttt tctcatttat aaggttaaat
     4861 aattctcata tatcaagcaa agtgacaggc gcccttaaat attctgacaa atgctctttc
     4921 cctaaactcc ccccataaaa aaacccgccg aagcgggttt ttacgttatt tgcggattaa
     4981 cgattactcg ttatcagaac cgcccagggg gcccgagctt aagactggcc gtcgttttac
     5041 aacacagaaa gagtttgtag aaacgcaaaa aggccatccg tcaggggcct tctgcttagt
     5101 ttgatgcctg gcagttccct actctcgcct tccgcttcct cgctcactga ctcgctgcgc
     5161 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5221 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5281 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5341 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5401 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5461 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5521 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5581 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5641 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5701 ggtgctacag agttcttgaa gtggtgggct aactacggct acactagaag aacagtattt
     5761 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5821 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5881 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5941 aacgacgcgc gcgtaactca cgttaaggga ttttggtcat gagcttgcgc cgtcccgtca
     6001 agtcagcgta atgctctgct taccaatgct taatcagtga ggcacctatc tcagcgatct
     6061 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6121 agggcttacc atctggcccc agcgctgcga tgataccgcg agaaccacgc tcaccggctc
     6181 cggatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6241 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6301 cagttaatag tttgcgcaac gttgttgcca tcgctacagg catcgtggtg tcacgctcgt
     6361 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     6421 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     6481 tggccgcagt gttatcactc atggttatgg cagcgctaca taattctctt actgtcatgc
     6541 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     6601 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6661 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6721 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6781 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6841 aaaagggaat aagggcgaca cggaaatgtt gaatactcat attcttcctt tttcaatatt
     6901 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6961 aaaataaaca aataggggtc agtgttacaa ccaattaacc aattctgaac attatcgcga
     7021 gcccatttat acctgaatat ggctcataac accccttgtt tgcctggcgg cagtagcgcg
     7081 gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt
     7141 gtggggactc cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca
     7201 gtcgaaagac tgggcctttc gcccgggcta attatggggt gtcgccctta ttcgactcta
     7261 tagtgaagtt cctattctct agaaagtata ggaacttctg aagtggggat ttaaatgcgg
     7321 ccgcgctgag ggtttaatcg acgaagcagc tgacggccag tgccaagctt aacgcgtacc
     7381 gggcccagta tatgttccgc agatgactgg agctctgcca tacgtgccct ctcaagcacc
     7441 atttgttcca tctacagaga ctagtcacca actagtctat caagactcac agggtacatt
     7501 gctgagacca actgaccaga ggcagggtag cggattgacg gctccatctc cttcacttac
     7561 aaggtctatt gaaagccctt tagcatcacc aagcggagaa tagattgtta agcttatttt
     7621 ttgtatactg ttttgtgata gcacgaagtt tttccacggt atcttgttaa aaatatatat
     7681 ttgtggcggg cttacctaca tcaaattaat aagagactaa ttataaacta aacacacaag
     7741 caagctactt tagggtaaaa gtttataaat gcttttgacg tataaacgtt gcttgtattt
     7801 attattacaa ttaaaggtgg atagaaaacc tagagactag ttagaaacta atctcaggtt
     7861 tgcgttaaac taaatcagag cccgagaggt taacagaacc tagaagggga ctagatatcc
     7921 gggtagggaa acaaaaaaaa aaacaagaca gccacatatt agggagacta gttagaagct
     7981 agttccagga ctaggaaaat aaaagacaat gataccacag tctagttgac aactagatag
     8041 attctagatt gaggccaaag tctctgagat ccaggttagt tgcaactaat actagttagt
     8101 atctagtctc ctataactct gaagctagaa taacttacta ctattatcct caccactgtt
     8161 cagctgcgca aacggagtga ttgcaaggtg ttcagagact agttattgac tagtcagtga
     8221 ctagcaataa ctaacaaggt attaacctac catgtctgcc atcaccctgc acttcctcgg
     8281 gctcagcagc cttttcctcc tcattttcat gctcattttc cttgtttaag actgtgacta
     8341 gtcaaagact agtccagaac cacaaaggag aaatgtctta ccactttctt cattgcttgt
     8401 ctcttttgca ttatccatgt ctgcaactag ttagagtcta gttagtgact agtccgacga
     8461 ggacttgctt gtctccggat tgttggagga actctccagg gcctcaagat ccacaacaga
     8521 gccttctaga agactggtca ataactagtt ggtctttgtc tgagtctgac ttacgaggtt
     8581 gcatactcgc tccctttgcc tcgtcaatcg atgagaaaaa gcgccaaaac tcgcaatatg
     8641 gctttgaacc acacggtgct gagactagtt agaatctagt cccaaactag cttggatagc
     8701 ttacctttgc cctttgcgtt gcgacaggtc ttgcagggta tggttccttt ctcaccagct
     8761 gatttagctg ccttgctacc ctcacggcgg atctgcataa agagtggcta gaggttataa
     8821 attagcactg atcctaggta cggggctgaa tgtaacttgc ctttcctttc tcatcgcgcg
     8881 gcaagacagg cttgctcaaa ttcctaccag tcacaggggt atgcacggcg tacggaccac
     8941 ttgaactagt cacagattag ttagcaacta gtctgcattg aatggctgta cttacgggcc
     9001 ctcgccattg tcctgatcat ttccagcttc accctcgttg ctgcaaagta gttagtgact
     9061 agtcaaggac tagttgaaat gggagaagaa actcacgaat tctcgacacc cttagtattg
     9121 tggtccttgg acttggtgct gctatatatt agctaataca ctagttagac tcacagaaac
     9181 ttacgcagct cgcttgcgct tcttggtagg agtcggggtt gggagaacag tgccttcaaa
     9241 caagccttca taccatgcta cttgactagt cagggactag tcaccaagta atctagatag
     9301 gacttgcctt tggcctccat cagttccttc atagtgggag gtccattgtg caatgtaaac
     9361 tccatgccgt gggagttctt atccttcaag tgcttgacca atatgtttct gttggcagag
     9421 ggaacctgtc aactagttaa taactagtca gaaactagta tagcagtaga ctcactgtac
     9481 gcttgaggca tcccttcact cggcagtaga cttcatatgg atggatatca ggcacgccat
     9541 tgtcgtcctg tggactagtc agtaactagg cttaaagcta gtcgggtcgg cttactatct
     9601 tgaaatccgg cagcgtaagc tccccgtcct taactgcctc gagatagtga cagtactctg
     9661 gggactttcg gagatcgtta tcgcgaatgc tcggcatact aatcgttgac tagtcttgga
     9721 ctagtcccga gcaaaaagga ttggaggagg aggaggaagg tgagagtgag acaaagagcg
     9781 aaataagagc ttcaaaggct atctctaagc agtatgaagg ttaagtatct agttcttgac
     9841 tagatttaaa agagatttcg actagttatg tacctggagt ttggatatag gaatgtgttg
     9901 tggtaacgaa atgtaagggg gaggaaagaa aaagtcggtc aagaggtaac tctaagtcgg
     9961 ccattccttt ttgggaggcg ctaaccataa acggcatggt cgacttagag ttagctcagg
    10021 gaatttaggg agttatctgc gaccaccgag gaacggcgga atgccaaaga atcccgatgg
    10081 agctctagct ggcggttgac aaccccacct tttggcgttt ctgcggcgtt gcaggcggga
    10141 ctggatactt cgtagaacca gaaaggcaag gcagaacgcg ctcagcaaga gtgttggaag
    10201 tgatagcatg atgtgccttg ttaactaggt caaaatctgc agtatgcttg atgttatcca
    10261 aagtgtgaga gaggaaggtc caaacataca cgattgggag agggcctagg tataagagtt
    10321 tttgagtaga acgcatgtga gcccagccat ctcgaggaga ttaaacacgg gccggcattt
    10381 gatggctatg ttagtacccc aatggaaacg gtgagagtcc agtggtcgca gataactccc
    10441 taaattccct gagctaactc taagtcgacc atgccgttta tggttagcgc ctcccaaaaa
    10501 ggaatggccg acttagagtt acctcttgac cgactttttc tttcctcccc cttacatttc
    10561 gttaccacaa cacattccta tatccaaact ccaggtacat aactagtcga aatctctttt
    10621 aaatctagtc aagaactaga tacttaacct tcatactgct tagagatagc ctttgaagct
    10681 cttatttcgc tctttgtctc actctcacct tcctcctcct cctccaatcc tttttgctcg
    10741 ggactagtcc aagactagtc aacgattagt atgccgagca ttcgcgataa cgatctccga
    10801 aagtccccag agtactgtca ctatctcgag gcagttaagg acggggagct tacgctgccg
    10861 gatttcaaga tagtaagccg acccgactag ctttaagcct agttactgac tagtccacag
    10921 gacgacaatg gcgtgcctga tatccatcca tatgaagtct actgccgagt gaagggatgc
    10981 ctcaagcgta cagtgagtct actgctatac tagtttctga ctagttatta actagttgac
    11041 aggttccctc tgccaacaga aacatattgg tcaagcactt gaaggacaag aactcccacg
    11101 gcatggagtt tacattgcac aatggacctc ccactatgaa ggaactgatg gaggccaaag
    11161 gcaagtccta tctagattac ttggtgacta gtccctgact agtcaagtag catggtatga
    11221 aggcttgttt gaaggcactg ttctcccaac cccgactcct accaagaagc gcaagcgagc
    11281 tgcgtaagtt tctgtgagtc taactagtgt attagctaat atatagcagc accaagtcca
    11341 aggaccacaa tactaagggt gtcgagaatt cgtgagtttc ttctcccatt tcaactagtc
    11401 cttgactagt cactaactac tttgcagcaa cgagggtgaa gctggaaatg atcaggacaa
    11461 tggcgagggc ccgtaagtac agccattcaa tgcagactag ttgctaacta atctgtgact
    11521 agttcaagtg gtccgtacgc cgtgcatacc cctgtgactg gtaggaattt gagcaagcct
    11581 gtcttgccgc gcgatgagaa aggaagggca agttacattc agccccgtac ctaggatcag
    11641 tgctaattta taacctctag ccactcttta tgcagatccg ccgtgagggt agcaaggcag
    11701 ctaaatcagc tggtgagaaa ggaaccatac cctgcaagac ctgtcgcaac gcaaagggca
    11761 aaggtaagct atccaagcta gtttgggact agattctaac tagtctcagc accgtgtggt
    11821 tcaaagccat attgcgagtt ttggcgcttt ttctcatcga ttgacgaggc aaagggagcg
    11881 agtatgcaac ctcgtaagtc agactcagac aaagaccaac tagttattga ccagtcttct
    11941 agaaggctct gttgtggatc ttgaggccct ggagagttcc tccaacaatc cggagacaag
    12001 caagtcctcg tcggactagt cactaactag actctaacta gttgcagaca tggataatgc
    12061 aaaagagaca agcaatgaag aaagtggtaa gacatttctc ctttgtggtt ctggactagt
    12121 ctttgactag tcacagtctt aaacaaggaa aatgagcatg aaaatgagga ggaaaaggct
    12181 gctgagcccg aggaagtgca gggtgatggc agacatggta ggttaatacc ttgttagtta
    12241 ttgctagtca ctgactagtc aataactagt ctctgaacac cttgcaatca ctccgtttgc
    12301 gcagctgaac agtggtgagg ataatagtag taagttattc tagcttcaga gttataggag
    12361 actagatact aactagtatt agttgcaact aacctggatc tcagagactt tggcctcaat
    12421 ctagaatcta tctagttgtc aactagactg tggtatcatt gtcttttatt ttcctagtcc
    12481 tggaactagc ttctaactag tctccctaat atgtggctgt cttgtttttt ttttttgttt
    12541 ccctacccgg atatctagtc cccttctagg ttctgttaac ctctcgggct ctgatttagt
    12601 ttaacgcaaa cctgagatta gtttctaact agtctctagg ttttctatcc acctttaatt
    12661 gtaataataa atacaagcaa cgtttatacg tcaaaagcat ttataaactt ttaccctaaa
    12721 gtagcttgct tgtgtgttta gtttataatt agtctcttat taatttgatg taggtaagcc
    12781 cgccacaaat atatattttt aacaagatac cgtggaaaaa cttcgtgcta tcacaaaaca
    12841 gtatacaaaa aataagct
//
