LOCUS       sequence-97424-e       11241 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses VP64 human MyoD and dsRedExpress2 in response to 
            doxycycline.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11241)
  AUTHORS   Kabadi AM, Thakore PI, Vockely CM, Ousterout DG, Gibson TM, Guilak 
            F, Reddy TE, Gersbach CA
  TITLE     Enhanced MyoD-Induced Transdifferentiation to a Myogenic Lineage by 
            Fusion to a Potent Transactivation Domain.
  JOURNAL   ACS Synth Biol. 2014 Dec 10.
  PUBMED    25494287
REFERENCE   2  (bases 1 to 11241)
  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 Synth 
            Biol. 2014 Dec 10."
COMMENT     Sequence Label: LV-TRE-VP64 human MyoD-T2A-dsRedExpress2
FEATURES             Location/Qualifiers
     source          1..11241
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    39..627
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(92..112)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(510..521)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             714..947
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1019..1140
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1056..1075)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1110..1129
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1159..1294
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1326..1345)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1327..1345)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1355..1372)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1355..1371)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1364..1386)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1516..1971
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1603..1622)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1813..1834
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2000..2104
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2105..2173,2174..2965)
                     /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(2323..2342)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3136..3724
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3625..3644
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3878..3895
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4012..4033
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4048..4053,4054..4071,4072..4078)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4086..4102
                     /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     4091..4113
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4110..4126
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4110..4126
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4145..4165
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4147..4165
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     LTR             4421..4601
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4645..4770
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5263..5496
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5681..5725
                     /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    5992..6109
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    join(6142..6160,6161..6183,6184..6202,6203..6225,6226..6244,
                     6245..6267,6268..6286,6287..6309,6310..6328,6329..6351,
                     6352..6370,6371..6393,6394..6412)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        6445..6482
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6479..6503
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             6672..6692
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6702..6851
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             6858..6884
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7875..7928
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             7929..8603
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(8109..8129)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     8327..8346
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     8521..8544
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     promoter        8694..9204
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     9123..9141
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             join(9243..9860,9861..9866,9867..9902,9903..9905,9906..9941,
                     9942..9944,9945..9980,9981..9989)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    join(10034..10608,10609..10611,10612..10620)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(10201..10218)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     10428..10447
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             10621..11220
                     /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(10621..10640)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     11117..11137
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 ctagcttctg cagcacgtgt ctagaaacgc tggtcgacaa tcaacctctg gattacaaaa
       61 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
      121 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
      181 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
      241 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
      301 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
      361 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
      421 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc
      481 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
      541 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
      601 ggatctccct ttgggccgcc tccccgcctg gaattcgagc tcggtacctt taagaccaat
      661 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
      721 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt
      781 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
      841 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
      901 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
      961 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa
     1021 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     1081 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     1141 tcatgtctgg ctctagctat cccgccccta actccgccca gttccgccca ttctccgccc
     1201 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     1261 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcgtcga gacgtaccca
     1321 attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg
     1381 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     1441 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     1501 atggcgaatg gcgcgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     1561 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     1621 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     1681 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     1741 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     1801 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     1861 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     1921 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt tcccaggtgg
     1981 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2041 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2101 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2161 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2221 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2281 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     2341 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2401 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     2461 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     2521 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     2581 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     2641 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     2701 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     2761 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     2821 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     2881 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     2941 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3001 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3061 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3121 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3181 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3241 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     3301 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3361 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3421 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3481 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3541 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3601 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3661 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3721 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3781 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3841 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     3901 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     3961 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     4021 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     4081 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     4141 gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct
     4201 tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa
     4261 ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta
     4321 ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc
     4381 agagatattg tatttaagtg cctagctcga tacaataaac gggtctctct ggttagacca
     4441 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     4501 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     4561 atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac
     4621 ctgaaagcga aagggaaacc agagctctct cgacgcagga ctcggcttgc tgaagcgcgc
     4681 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
     4741 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
     4801 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
     4861 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
     4921 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
     4981 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
     5041 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
     5101 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     5161 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     5221 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     5281 gttcttggga gcagcaggaa gcactatggg cgcagcctca atgacgctga cggtacaggc
     5341 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     5401 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     5461 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     5521 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     5581 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     5641 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     5701 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     5761 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     5821 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     5881 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     5941 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcggttaa cttttaaaag
     6001 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     6061 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaatttta tcgatcacga
     6121 gactagcctc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
     6181 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata
     6241 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
     6301 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca
     6361 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa
     6421 agtcgagctc ggtacccggg tcgagtaggc gtgtacggtg ggaggcctat ataagcagag
     6481 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     6541 gaagacaccg ggaccgatcc agcctccgcg gccccgaatt cgagctcggt acccggggat
     6601 cctctagtca gctgacgcgt ttggttatgc atgccaccat gaaaactagt ggacaggccg
     6661 gccaggctag cccgaaaaag aaacgcaaag ttgggcgcgc cgacgcgctg gacgatttcg
     6721 atctcgacat gctgggttct gatgccctcg atgactttga cctggatatg ttgggaagcg
     6781 acgcattgga tgactttgat ctggacatgc tcggctccga tgctctggac gatttcgatc
     6841 tcgatatgtt aattaactac ccgtacgacg ttccggacta cgcttctgga ggcagtgggg
     6901 gcggatcaat ggagctactg tcgccaccgc tccgcgacgt agacctgacg gcccccgacg
     6961 gctctctctg ctcctttgcc acaacggacg acttctatga cgacccgtgt ttcgactccc
     7021 cggacctgcg cttcttcgaa gacctggacc cgcgcctgat gcacgtgggc gcgctcctga
     7081 aacccgaaga gcactcgcac ttccccgcgg cggtgcaccc ggccccgggc gcacgtgagg
     7141 acgagcatgt gcgcgcgccc agcgggcacc accaggcggg ccgctgccta ctgtgggcct
     7201 gcaaggcgtg caagcgcaag accaccaacg ccgaccgccg caaggccgcc accatgcgcg
     7261 agcggcgccg cctgagcaaa gtaaatgagg cctttgagac actcaagcgc tgcacgtcga
     7321 gcaatccaaa ccagcggttg cccaaggtgg agatcctgcg caacgccatc cgctatatcg
     7381 agggcctgca ggctctgctg cgcgaccagg acgccgcgcc ccctggcgcc gcagccgcct
     7441 tctatgcgcc gggcccgctg cccccgggcc gcggcggcga gcactacagc ggcgactccg
     7501 acgcgtccag cccgcgctcc aactgctccg acggcatgat ggactacagc ggccccccga
     7561 gcggcgcccg gcggcggaac tgctacgaag gcgcctacta caacgaggcg cccagcgaac
     7621 ccaggcccgg gaagagtgcg gcggtgtcga gcctagactg cctgtccagc atcgtggagc
     7681 gcatctccac cgagagccct gcggcgcccg ccctcctgct ggcggacgtg ccttctgagt
     7741 cgcctccgcg caggcaagag gctgccgccc ccagcgaggg agagagcagc ggcgacccca
     7801 cccagtcacc ggacgccgcc ccgcagtgcc ctgcgggtgc gaaccccaac ccgatatacc
     7861 aggtgctcat gcatgagggc agaggaagtc ttctaacatg cggtgacgtg gaggagaatc
     7921 ccggccctat ggatagcact gagaacgtca tcaagccctt catgcgcttc aaggtgcaca
     7981 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggcaagccct
     8041 acgagggcac ccagaccgcc aagctgcagg tgaccaaggg cggccccctg cccttcgcct
     8101 gggacatcct gtccccccag ttccagtacg gctccaaggt gtacgtgaag caccccgccg
     8161 acatccccga ctacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     8221 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcacct
     8281 tcatctacca cgtgaagttc atcggcgtga acttcccctc cgacggcccc gtaatgcaga
     8341 agaagactct gggctgggag ccctccaccg agcgcctgta cccccgcgac ggcgtgctga
     8401 agggcgagat ccacaaggcg ctgaagctga agggcggcgg ccactacctg gtggagttca
     8461 agtcaatcta catggccaag aagcccgtga agctgcccgg ctactactac gtggactcca
     8521 agctggacat cacctcccac aacgaggact acaccgtggt ggagcagtac gagcgcgccg
     8581 aggcccgcca ccacctgttc cagactcacg gtatggacga attgtacaag cacgacgaat
     8641 tgtaagctag cgcggccgca tcgataagct tgtggacgat atcgaattcg cacggggttg
     8701 gggttgcgcc ttttccaagg cagccctggg tttgcgcagg gacgcggctg ctctgggcgt
     8761 ggttccggga aacgcagcgg cgccgaccct gggtctcgca cattcttcac gtccgttcgc
     8821 agcgtcaccc ggatcttcgc cgctaccctt gtgggccccc cggcgacgct tcctgctccg
     8881 cccctaagtc gggaaggttc cttgcggttc gcggcgtgcc ggacgtgaca aacggaagcc
     8941 gcacgtctca ctagtaccct cgcagacgga cagcgccagg gagcaatggc agcgcgccga
     9001 ccgcgatggg ctgtggccaa tagcggctgc tcagcggggc gcgccgagag cagcggccgg
     9061 gaaggggcgg tgcgggaggc ggggtgtggg gcggtagtgt gggccctgtt cctgcccgcg
     9121 cggtgttccg cattctgcaa gcctccggag cgcacgtcgg cagtcggctc cctcgttgac
     9181 cgaatcaccg acctctctcc ccagggggga tccgtttatc tgcagaattc gcccttgacg
     9241 tcatgtctag actggacaag agcaaagtca taaacggcgc tctggaatta ctcaatggag
     9301 tcggtatcga aggcctgacg acaaggaaac tcgctcaaaa gctgggagtt gagcagccta
     9361 ccctgtactg gcacgtgaag aacaagcggg ccctgctcga tgccctgcca atcgagatgc
     9421 tggacaggca tcatacccac ttctgccccc tggaaggcga gtcatggcaa gactttctgc
     9481 ggaacaacgc caagtcattc cgctgtgctc tcctctcaca tcgcgacggg gctaaagtgc
     9541 atctcggcac ccgcccaaca gagaaacagt acgaaaccct ggaaaatcag ctcgcgttcc
     9601 tgtgtcagca aggcttctcc ctggagaacg cactgtacgc tctgtccgcc gtgggccact
     9661 ttacactggg ctgcgtattg gaggaacagg agcatcaagt agcaaaagag gaaagagaga
     9721 cacctaccac cgattctatg cccccacttc tgagacaagc aattgagctg ttcgaccggc
     9781 agggagccga acctgccttc cttttcggcc tggaactaat catatgtggc ctggagaaac
     9841 agctaaagtg cgaaagcggc gggccggccg acgcccttga cgattttgac ttagacatgc
     9901 tcccagccga tgcccttgac gactttgacc ttgatatgct gcctgctgac gctcttgacg
     9961 attttgacct tgacatgctc cccgggtaac ggaccggacg tcaagggcga attccagcac
    10021 actgggaatt ccgcccctct ccctcccccc cccctaacgt tactggccga agccgcttgg
    10081 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
    10141 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
    10201 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
    10261 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
    10321 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
    10381 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
    10441 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
    10501 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
    10561 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc
    10621 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
    10681 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
    10741 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
    10801 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
    10861 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
    10921 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
    10981 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
    11041 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
    11101 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
    11161 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
    11221 ccagcacact ggcggccgtt a
//
