LOCUS       sequence-97416-e       11235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses VP64 mouse MyoD and dsRedExpress2 in response to 
            doxycycline.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11235)
  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 11235)
  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 mouse MyoD-T2A-dsRedExpress2
FEATURES             Location/Qualifiers
     source          1..11235
                     /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             7869..7922
                     /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             7923..8597
                     /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(8103..8123)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     8321..8340
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     8515..8538
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     promoter        8688..9198
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     9117..9135
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             join(9237..9854,9855..9860,9861..9896,9897..9899,9900..9935,
                     9936..9938,9939..9974,9975..9983)
                     /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(10028..10602,10603..10605,10606..10614)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(10195..10212)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     10422..10441
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             10615..11214
                     /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(10615..10634)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     11111..11131
                     /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 ggagcttcta tcgccgccac tccgggacat agacttgaca ggccccgacg
     6961 gctctctctg ctcctttgag acagcagacg acttctatga tgatccgtgt ttcgactcac
     7021 cagacctgcg cttttttgag gacctggacc cgcgcctggt gcacgtggga gccctcctga
     7081 aaccggagga gcacgcacac ttctctactg cggtgcaccc aggcccaggc gctcgtgagg
     7141 atgagcatgt gcgcgcgccc agcgggcacc accaggcggg tcgctgcttg ctgtgggcct
     7201 gcaaggcgtg caagcgcaag accaccaacg ctgatcgccg caaggccgcc accatgcgcg
     7261 agcgccgccg cctgagcaaa gtgaatgagg ccttcgagac gctcaagcgc tgcacgtcca
     7321 gcaacccgaa ccagcggcta cccaaggtgg agatcctgcg caacgccatc cgctacatcg
     7381 aaggtctgca ggctctgctg cgcgaccagg acgccgcgcc ccctggcgcc gctgccttct
     7441 acgcacctgg accgctgccc ccaggccgtg gcagcgagca ctacagtggc gactcagacg
     7501 cgtccagccc gcgctccaac tgctctgatg gcatgatgga ttacagcggc cccccaagcg
     7561 gcccccggcg gcagaatggc tacgacaccg cctactacag tgaggcggtg cgcgagtcca
     7621 ggccagggaa gagtgcggct gtgtcgagcc tcgactgcct gtccagcata gtggagcgca
     7681 tctccacaga cagccccgct gcgcctgcgc tgcttttggc agatgcacca ccagagtcgc
     7741 ctccgggtcc gccagagggg gcatccctaa gcgacacaga acagggaacc cagaccccgt
     7801 ctcccgacgc cgcccctcag tgtcctgcag gctcaaaccc caatgcgatt tatcaggtgc
     7861 ttatgcatga gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc
     7921 ctatggatag cactgagaac gtcatcaagc ccttcatgcg cttcaaggtg cacatggagg
     7981 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggcaag ccctacgagg
     8041 gcacccagac cgccaagctg caggtgacca agggcggccc cctgcccttc gcctgggaca
     8101 tcctgtcccc ccagttccag tacggctcca aggtgtacgt gaagcacccc gccgacatcc
     8161 ccgactacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     8221 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc accttcatct
     8281 accacgtgaa gttcatcggc gtgaacttcc cctccgacgg ccccgtaatg cagaagaaga
     8341 ctctgggctg ggagccctcc accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg
     8401 agatccacaa ggcgctgaag ctgaagggcg gcggccacta cctggtggag ttcaagtcaa
     8461 tctacatggc caagaagccc gtgaagctgc ccggctacta ctacgtggac tccaagctgg
     8521 acatcacctc ccacaacgag gactacaccg tggtggagca gtacgagcgc gccgaggccc
     8581 gccaccacct gttccagact cacggtatgg acgaattgta caagcacgac gaattgtaag
     8641 ctagcgcggc cgcatcgata agcttgtgga cgatatcgaa ttcgcacggg gttggggttg
     8701 cgccttttcc aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc
     8761 gggaaacgca gcggcgccga ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc
     8821 acccggatct tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta
     8881 agtcgggaag gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt
     8941 ctcactagta ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga
     9001 tgggctgtgg ccaatagcgg ctgctcagcg gggcgcgccg agagcagcgg ccgggaaggg
     9061 gcggtgcggg aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt
     9121 tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc
     9181 accgacctct ctccccaggg gggatccgtt tatctgcaga attcgccctt gacgtcatgt
     9241 ctagactgga caagagcaaa gtcataaacg gcgctctgga attactcaat ggagtcggta
     9301 tcgaaggcct gacgacaagg aaactcgctc aaaagctggg agttgagcag cctaccctgt
     9361 actggcacgt gaagaacaag cgggccctgc tcgatgccct gccaatcgag atgctggaca
     9421 ggcatcatac ccacttctgc cccctggaag gcgagtcatg gcaagacttt ctgcggaaca
     9481 acgccaagtc attccgctgt gctctcctct cacatcgcga cggggctaaa gtgcatctcg
     9541 gcacccgccc aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg ttcctgtgtc
     9601 agcaaggctt ctccctggag aacgcactgt acgctctgtc cgccgtgggc cactttacac
     9661 tgggctgcgt attggaggaa caggagcatc aagtagcaaa agaggaaaga gagacaccta
     9721 ccaccgattc tatgccccca cttctgagac aagcaattga gctgttcgac cggcagggag
     9781 ccgaacctgc cttccttttc ggcctggaac taatcatatg tggcctggag aaacagctaa
     9841 agtgcgaaag cggcgggccg gccgacgccc ttgacgattt tgacttagac atgctcccag
     9901 ccgatgccct tgacgacttt gaccttgata tgctgcctgc tgacgctctt gacgattttg
     9961 accttgacat gctccccggg taacggaccg gacgtcaagg gcgaattcca gcacactggg
    10021 aattccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
    10081 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
    10141 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
    10201 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
    10261 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
    10321 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
    10381 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
    10441 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
    10501 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
    10561 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatgacc
    10621 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
    10681 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
    10741 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
    10801 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
    10861 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
    10921 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
    10981 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
    11041 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
    11101 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
    11161 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaccagca
    11221 cactggcggc cgtta
//
