LOCUS       sequence-475682-       11883 bp    DNA     circular SYN 11-JUN-2025
DEFINITION  Tetracycline-inducible expression of Isl1 and Lhx3 for direct 
            reprogramming of canine fibroblasts to induced-motor neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11883)
  TITLE     Protocol for direct reprogramming of canine fibroblasts to 
            induced-motor neurons
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11883)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 11, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pLVX-UbC-rtTA-Isl1_2A_Lhx3-BSD
FEATURES             Location/Qualifiers
     source          1..11883
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     LTR             1119..1752
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    1959..2547
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2065..2076
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2474..2494
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     promoter        complement(3003..3502)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3051..3070)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     3444..3467
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     regulatory      4729..4738
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4745..4798)
                     /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"
     regulatory      5073..5082
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      5869..5878
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5893..6207)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     primer_bind     complement(5896..5920)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    5967..5985
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6003..6021
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6038..6056
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6074..6092
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6110..6128
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6145..6163
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6181..6199
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        6245..7456
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     7293..7312
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(7466..8083,8084..8089,8090..8125,8126..8128,8129..8164,
                     8165..8167,8168..8203,8204..8215)
                     /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
                     DALDDFDLDMLPADALDDFDLDMLPGN"
     misc_feature    8256..8371
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     CDS             complement(8634..8678)
                     /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    8863..9096
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    9593..9718
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             9765..10398
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    10566..10700
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10590..10609)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     10644..10663
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        10748..10852
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10853..10921,10922..11713)
                     /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(11071..11090)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt
     1021 cgacggtaaa cgctagcaat gtcaaggcct ctcactctct gatattcatt tctttgcaag
     1081 ttataaatac tgaataataa gatgacatga actactactg ctagagattt tccacactga
     1141 ctaaaagggt ctgagggatc tctagttacc agagtcacac aacagacggg cacacactac
     1201 ttgaagcact caaggcaagc tttattgagg cttaagcagt gggttcccta gttagccaga
     1261 gagctcccag gctcagatct ggtctaacca gagagaccca gtacaggcaa aaagcagctg
     1321 cttatatgca ggatctgagg gctcgccact ccccagtccc gcccaggcca cgcctccctg
     1381 gaaagtcccc agcggaaagt cccttgtagc aagctcgata tcagcagttc ttgaagtact
     1441 ccggatgcag ctctcgggcc acgtgatgaa atgctaggcg gctgtcaaac ctccactcta
     1501 acacttctct ctccgggtca tccatcccat gcaggctcac agggtgtaac aagctggtgt
     1561 tctctccttt attggcctct tctaccttat ctggctcaac tggtactagc ttgtagcacc
     1621 atccaaaggt cagtggatat ctgacccctg gccctggtgt gtagttctgc taatcaggga
     1681 agtagccttg tgtgtggtag atccacagat caaggatatc ttgtcttcgt tgggagtgaa
     1741 ttagcccttc cagtcccctc ttttctttta aaaagtggct aagatctaca gctgccttgt
     1801 aagtcattgg tcttaaaggt acctgaggtg tgactggaaa acccacctcc tcctcctctt
     1861 gtgcttctag ccaggcacaa tcagcattgg tagctgctgt attgctactt gtgattgctc
     1921 catgtttttc taggtctcga ctgcagaatt aattccaggc ggggaggcgg cccaaaggga
     1981 gatccgactc gtctgagggc gaaggcgaag acgcggaaga ggccgcagag ccggcagcag
     2041 gccgcgggaa ggaaggtccg ctggattgag ggccgaaggg acgtagcaga aggacgtccc
     2101 gcgcagaatc caggtggcaa cacaggcgag cagccatgga aaggacgtca gcttccccga
     2161 caacaccacg gaattgtcag tgcccaacag ccgagcccct gtccagcagc gggcaaggca
     2221 ggcggcgatg agttccgccg tggcaatagg gagggggaaa gcgaaagtcc cggaaaggag
     2281 ctgacaggtg gtggcaatgc cccaaccagt gggggttgcg tcagcaaaca cagtgcacac
     2341 cacgccacgt tgcctgacaa cgggccacaa ctcctcataa agagacagca accaggattt
     2401 atacaaggag gagaaaatga aagccatacg ggaagcaata gcatgataca aaggcattaa
     2461 agcagcgtat ccacatagcg taaaaggagc aacatagtta agaataccag tcaatctttc
     2521 acaaattttg taatccagag gttgattgtt ccagacgcgc cttaatttcg ggtatatttg
     2581 agtggaatga gttcttcaat cgtagttttg actaacttgc cattcatttc tattaacaca
     2641 aaacaatctg gtgcatagtc tgaaatcaac tccctacaca taccacaagg acttaccact
     2701 cgaatacttc tatctacttc gtcagaataa gggtgtctaa cagctacaat cgtgtcaaaa
     2761 tccttttgtc cattcgaaac tgcactacca atcgcaatgg cttctgcaca aacagttact
     2821 cgtcctatat acgcttcaat atgtactgcc gaaatgattt ctcctgtttt cgtacgaatt
     2881 gccgctccca catgatgttt attatcctca taaagcattg taatcttctc tgtcgctact
     2941 tctactaatt ctagatcctg ctgggagatg ttgaaggtct tcatggtggg tgggctgcag
     3001 gtcgaaaggc ccggagatga ggaagaggag aacagcgcgg cagacgtgcg cttttgaagc
     3061 gtgcagaatg ccgggcctcc ggaggacctt cgggcgcccg ccccgcccct gagcccgccc
     3121 ctgagcccgc ccccggaccc accccttccc agcctctgag cccagaaagc gaaggagcaa
     3181 agctgctatt ggccgctgcc ccaaaggcct acccgcttcc attgctcagc ggtgctgtcc
     3241 atctgcacga gactagtgag acgtgctact tccatttgtc acgtcctgca cgacgcgagc
     3301 tgcggggcgg gggggaactt cctgactagg ggaggagtag aaggtggcgc gaaggggcca
     3361 ccaaagaacg gagccggttg gcgcctaccg gtggatgtgg aatgtgtgcg aggccagagg
     3421 ccacttgtgt agcgccaagt gcccagcggg gctgctaaag cgcatgctcc agactgcctt
     3481 gggaaaagcg cctcccctac ccggtagaat tcactccctc gagttagaac tgagcgtggt
     3541 ctacctcatc cagccaggag gcggggctgg cagggaagtc ggggtaaccc ccgctgctcc
     3601 ccgtggatag gtcagaactg ggtccgttcc ctgccagcac cctcatgggt gggggcccgc
     3661 cgggggctcc cgagggcaca aggcccaagc tggtgtctgg gtacaccagg ctggagagga
     3721 ggggctgggg gccagggagg ctctgcgggg cggcggggga tggggggaca ccgtaggggc
     3781 tgccgggacg cagctctcgg tactgctctg ggcctgccag gcctccatgc tccagggaga
     3841 agttgcccag ggctcccgag ggccggccca aggcctgggt gggttccccc aagctcccgt
     3901 agaggccatt ggccgggccc atttccgcca aggaaggctc atcggggaag gagacctcag
     3961 cgtcgctgtc ctgcccctcc tgaacgctgt ccttgtccga cttggagccg ccgcgggagc
     4021 gcttcatgtt gcggaaatac tgcccccagc gctgccggcc ggcgtccttc ttcagcctct
     4081 tctccttggc ccggcggttc tggaaccaaa cctgcaccac gcgcatgtcc aggcccgtct
     4141 cggacgagag ctgctcgcgc acgtggcgcg ccggcttggg cgaggtgttg taagcgctct
     4201 tcagcgtctc cagctgcttg gcggtgatgg tcgtgcgcgg ccgcttggcc gtggcctcgg
     4261 cctctcgctg cttggcggtt tcgtagtccg ccttgcacac gagccggctg tcctccatga
     4321 ggtagaactc gtcgcccgtg gccagctgcc gcttgcacac gacgcaggca aagcagtgca
     4381 ggtggtacac gaagtcctgg gcgcggcgca ccacctgcgt gggcgggatg cccagctggc
     4441 acgcggcgca cttggtcccg aagcgcttga aaaagtcgtc cttgcagtaa acgctctccc
     4501 ctcggctgaa gcagcgctcg gccagtggcg tgtggcagtc gctgcacttg agacacttgc
     4561 tgtgccagtg gcggtccaga gccttgagga tgaagcggtc caggatgtgc tggtcacagc
     4621 cagcgcacag cgggatctct cggcgcaggt ccgccctccg cgccagcagt gctagcagca
     4681 ggtcgcctcc cgccgactcc cgggccgggc ccagctcccc gcgcgcctcc atggtggctc
     4741 tagcgggtcc aggattctct tcgacgtctc cgcaggtcag aagacttcct ctcccctcgt
     4801 aggatctgag tccggacttg taccttgcct caataggact ggctaccatg ctgttaggtg
     4861 tatctggaag ttgagaggac attgatgcta cttcactgcc agtggaatta gagcccggtc
     4921 ctccttctga aaaattgacc agttgctgaa aagcaggctg atctatgtca ctctgcaagg
     4981 cgaagtcgct cagtactttc caaggtggct ggtaactttg tacttccact gggttagcct
     5041 gtaagccacc gtcgtgtctc tctggactgg cagccaccat gggagttcct gtcatcccct
     5101 ggatattagt tttgtcattg ggctgctgct gctggagttg cttcatcatg atgcttcgct
     5161 tcttgtcctt gcaccgcttg ttttgaaacc agacccggat cacacgggga ctgaggcccg
     5221 tcatctctac cagttgctcc ttcatgagcg catctggccg cgggtttgcg gcgtagcagg
     5281 tccgcaaggt gtgcagctgc ttctcgttca gcacagtccg cacgcgggtg gtcttctccg
     5341 gctgcttgtg gacgtggggc cgcagggccg gctgcctggc ggagatgggc tccgctgcca
     5401 tttgcagtgg ccgcgctgga tgcaggggac tgagcgggtc gccagcgcct agactggccc
     5461 tctccaccac atcgtggtct gctcggcaga agagaccgtc ctcccgaagc gcaaattcgt
     5521 ccccagggat gagctggcgg ctgcaggcca cacagcggaa acactcgatg tgatacacct
     5581 tggagcgggc acgcatcacg aagtcgttct tgctgaagcc gatgctgcac ttggcgcatt
     5641 tgatcccgta caacctgata taatctcttt tacagtaggt tttcccatcc ctaacaaagc
     5701 atgtacagct ctcgtccaaa tactgattac actccgcaca tttcaaacat gccgcatgcc
     5761 attccaaatc cggagaaacc ctcagaatat actgatcgtg aatctgattg ccgcaaccaa
     5821 cacataggga aatcagacgt tttttttttg gtggatctcc catgtctccc atggtggcag
     5881 gatccttctc caggcgatct gacggttcac taaacgagct ctgcttatat aggcctccca
     5941 ccgtacacgc ctacctcgac atacgttctc tatcactgat agggagtaaa ctcgacatac
     6001 gttctctatc actgataggg ataaactcga catacgttct ctatcactga tagggagtaa
     6061 actcgacata cgttctctat cactgatagg gagtaaactc gacatacgtt ctctatcact
     6121 gatagggagt aaactcgaca tcgttctcta tcactgatag ggagtaaact cgacatacgt
     6181 tctctatcac tgatagggag taaactcgag tgaagacgaa agggcctcat cgatgcgccg
     6241 atctggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcct cacggcgagc
     6301 gctgccacgt cagacgaagg gcgcagcgag cgtcctgatc cttccgcccg gacgctcagg
     6361 acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt
     6421 ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc
     6481 gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc
     6541 cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt
     6601 gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg
     6661 ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg tgtggagaga
     6721 ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg ggttgggggg
     6781 agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt gaggcgggct
     6841 gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa ggtcttgagg
     6901 ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctggggcac catctgggga
     6961 ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg
     7021 cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg
     7081 tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg
     7141 tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct
     7201 gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg
     7261 gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc
     7321 ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg
     7381 gtcaatatgt aattttcagt gttagactag taaattgtcc gctaaattct ggccgttttt
     7441 ggcttttttg ttagacggat ccaccatgtc tagactggac aagagcaaag tcataaacgg
     7501 cgctctggaa ttactcaatg gagtcggtat cgaaggcctg acgacaagga aactcgctca
     7561 aaagctggga gttgagcagc ctaccctgta ctggcacgtg aagaacaagc gggccctgct
     7621 cgatgccctg ccaatcgaga tgctggacag gcatcatacc cacttctgcc ccctggaagg
     7681 cgagtcatgg caagactttc tgcggaacaa cgccaagtca ttccgctgtg ctctcctctc
     7741 acatcgcgac ggggctaaag tgcatctcgg cacccgccca acagagaaac agtacgaaac
     7801 cctggaaaat cagctcgcgt tcctgtgtca gcaaggcttc tccctggaga acgcactgta
     7861 cgctctgtcc gccgtgggcc actttacact gggctgcgta ttggaggaac aggagcatca
     7921 agtagcaaaa gaggaaagag agacacctac caccgattct atgcccccac ttctgagaca
     7981 agcaattgag ctgttcgacc ggcagggagc cgaacctgcc ttccttttcg gcctggaact
     8041 aatcatatgt ggcctggaga aacagctaaa gtgcgaaagc ggcgggccgg ccgacgccct
     8101 tgacgatttt gacttagaca tgctcccagc cgatgccctt gacgactttg accttgatat
     8161 gctgcctgct gacgctcttg acgattttga ccttgacatg ctcccgggta actaatcgat
     8221 aaactggatc tctgctgtcc ctgtaataaa cccgaaaatt ttgaattttt gtaatttgtt
     8281 tttgtaattc tttagtttgt atgtctgttg ctattatgtc tactattctt tcccctgcac
     8341 tgtacccccc aatcccccct tttcttttaa aggcgatacc gtcgagatcc gttcactaat
     8401 cgaatggatc tgtctctgtc tctctctcca ccttcttctt ctattccttc gggcctgtcg
     8461 ggtcccctcg gggttgggag gtgggtctga aacgataatg gtgaatatcc ctgcctaact
     8521 ctattcacta tagaaagtac agcaaaaact attcttaaac ctaccaagcc tcctactatc
     8581 attatgaata attttatata ccacagccaa tttgttatgt taaaccaatt ccacaaactt
     8641 gcccatttat ctaattccaa taattcttgt tcattctttt cttgctggtt ttgcgattct
     8701 tcaattaagg agtgtattaa gcttgtgtaa ttgttaattt ctctgtccca ctccatccag
     8761 gtcgtgtgat tccaaatctg ttccagagat ttattactcc aactagcatt ccaaggcaca
     8821 gcagtggtgc aaatgagttt tccagagcaa ccccaaatcc ccaggagctg ttgatccttt
     8881 aggtatcttt ccacagccag gattcttgcc tggagctgct tgatgcccca gactgtgagt
     8941 tgcaacagat gctgttgcgc ctcaatagcc ctcagcaaat tgttctgctg ctgcactata
     9001 ccagacaata attgtctggc ctgtaccgtc agcgtcattg acgctgcgcc catagtgctt
     9061 cctgctgctc ccaagaaccc aaggaacaaa gctcctattc ccactgctct tttttctctc
     9121 tgcaccactc ttctctttgc cttggtgggt gctactccta atggttcaat ttttactact
     9181 ttatatttat ataattcact tctccaattg tccctcatat ctcctcctcc aggtctgaag
     9241 atcagcggcc ggccgcttgc tgtgcggtgg tcttactttt gttttgctct tcctctatct
     9301 tgtctaaagc ttccttggtg tcttttatct ctatcctttg atgcacacaa tagagggttg
     9361 ctactgtatt atataatgat ctaagttctt ctgatcctgt ctgaagggat ggttgtagct
     9421 gtcccagtat ttgtctacag ccttctgatg tttctaacag gccaggatta actgcgaatc
     9481 gttctagctc cctgcttgcc catactatat gttttaattt atattttttc tttccccctg
     9541 gccttaaccg aattttttcc catcgcgatc taattctccc ccgcttaata ctgacgctct
     9601 cgcacccatc tctctccttc tagcctccgc tagtcaaaat ttttggcgta ctcaccagtc
     9661 gccgcccctc gcctcttgcc gtgcgcgctt cagcaagccg agtcctgcgt cgagagagct
     9721 cctctggttt ccctttcgct ttcaagtccc tgttcgggcg ccactgctag agattttcca
     9781 cactgactaa aagggtctga gggatctcta gttaccagag tcacacaaca gacgggcaca
     9841 cactacttga agcactcaag gcaagcttta ttgaggctta agcagtgggt tccctagtta
     9901 gccagagagc tcccaggctc agatctggtc taaccagaga gacccagtac aggcaaaaag
     9961 cagctgctta tatgcaggat ctgagggctc gccactcccc agtcccgccc aggccacgcc
    10021 tccctggaaa gtccccagcg gaaagtccct tgtagcaagc tcgatatcag cagttcttga
    10081 agtactccgg atgcagctct cgggccacgt gatgaaatgc taggcggctg tcaaacctcc
    10141 actctaacac ttctctctcc gggtcatcca tcccatgcag gctcacaggg tgtaacaagc
    10201 tggtgttctc tcctttattg gcctcttcta ccttatctgg ctcaactggt actagcttgt
    10261 agcaccatcc aaaggtcagt ggatatctga cccctggccc tggtgtgtag ttctgctaat
    10321 cagggaagta gccttgtgtg tggtagatcc acagatcaag gatatcttgt cttctttggg
    10381 agtgaattag cccttccaac tactaagttt gtagtacata tttaacaaat acaatttctt
    10441 taaaatgaaa ataattcaga ggaatcacag gtttagagta aatgaaacca caggtaattg
    10501 gcagtggtaa tagggtatgg ggtgggaagt ttgggatgat tttggttagc ttgagttatc
    10561 cagttgatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
    10621 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
    10681 taagctgcaa taaacaagtt gatctcccga tccgtcgacg tcaggtggca cttttcgggg
    10741 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
    10801 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
    10861 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
    10921 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
    10981 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
    11041 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
    11101 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
    11161 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
    11221 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
    11281 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
    11341 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
    11401 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
    11461 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
    11521 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
    11581 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
    11641 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
    11701 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
    11761 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
    11821 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
    11881 ttc
//
