LOCUS       sequence-213501-        9445 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-On construct to induce expression of tandem eGFP-mCherry tagged 
            TOM20MTS (mitochondria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9445)
  AUTHORS   Liang JR, Lingeman E, Ahmed S, Corn JE
  TITLE     Atlastins remodel the endoplasmic reticulum for selective autophagy.
  JOURNAL   J Cell Biol. 2018 Aug 24. pii: jcb.201804185. doi: 
            10.1083/jcb.201804185.
  PUBMED    30143524
REFERENCE   2  (bases 1 to 9445)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Cell 
            Biol. 2018 Aug 24. pii: jcb.201804185. doi: 10.1083/jcb.201804185."
COMMENT     Sequence Label: TOM20MTS-mCherry-EGFP-Tet-On
FEATURES             Location/Qualifiers
     source          1..9445
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             24..68
                     /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    281..398
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        452..766
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    460..478
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    496..514
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    531..549
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    567..585
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    603..621
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    638..656
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    674..692
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     739..763
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      783..792
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             903..1610
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1048..1066)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1316..1335
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    1638..1662
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1671..1680
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1677..1679,1680..1682,1683..2396)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1677..1679,1680..1682,1683..2393)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1722..1743)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1983..2002)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2330..2351
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        2424..2933
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2853..2871
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2943..3734
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2997..3016)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3607..3626
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             3744..3797
                     /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             join(3798..4415,4416..4421,4422..4457,4458..4460,4461..4496,
                     4497..4499,4500..4535,4536..4544)
                     /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
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(4584..4604)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5002..5013)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5218..5451
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5523..5644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5560..5579)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5614..5633
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5684..5819
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5819..5890
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5891..5959,5960..6751)
                     /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(6109..6128)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6922..7510
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7411..7430
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7664..7681
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7798..7819
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7834..7839,7840..7857,7858..7864)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7872..7888
                     /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     7877..7899
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7896..7912
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7896..7912
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7931..7951
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7933..7951
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7979..8205
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8206..8386
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8433..8558
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9051..9284
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
       61 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
      121 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
      181 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
      241 aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
      301 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
      361 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
      421 gagatccact ttggccgcga atcgatatgt cgagtttact ccctatcagt gatagagaac
      481 gtatgtcgag tttactccct atcagtgata gagaacgatg tcgagtttac tccctatcag
      541 tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt
      601 actccctatc agtgatagag aacgtatgtc gagtttatcc ctatcagtga tagagaacgt
      661 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg
      721 ggaggcctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gaattggcta
      781 gcgccaccat ggtgggtcgg aacagcgcca tcgccgccgg tgtatgcggg gcccttttca
      841 ttgggtactg catctacttc gaccgcaaaa gacgaagtgg tggatctgga ggttctggtg
      901 gaatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
      961 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     1021 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     1081 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     1141 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     1201 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     1261 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     1321 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     1381 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     1441 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     1501 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     1561 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag tccggactca
     1621 gatccgctgg cccaggaaca agtttgtaca aaaaagcagg ctttccggtc gccaccatgg
     1681 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1741 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1801 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1861 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     1921 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1981 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     2041 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     2101 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     2161 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     2221 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2281 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2341 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtagtccg
     2401 gactcagatc cggtaccgga tccggggttg gggttgcgcc ttttccaagg cagccctggg
     2461 tttgcgcagg gacgcggctg ctctgggcgt ggttccggga aacgcagcgg cgccgaccct
     2521 gggtctcgca cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc cgctaccctt
     2581 gtgggccccc cggcgacgct tcctgctccg cccctaagtc gggaaggttc cttgcggttc
     2641 gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca ctagtaccct cgcagacgga
     2701 cagcgccagg gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa tagcggctgc
     2761 tcagcagggc gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc ggggtgtggg
     2821 gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg cattctgcaa gcctccggag
     2881 cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg acctcttccc cagcaattca
     2941 ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3001 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3061 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3121 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3181 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3241 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3301 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     3361 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     3421 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     3481 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     3541 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     3601 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     3661 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     3721 ttgacgagtt cttccccggt gccgaaggta gaggttctct cctcacttgt ggtgatgttg
     3781 aagaaaaccc tggtccaatg tctagactgg acaagagcaa agtcataaac ggagctctgg
     3841 aattactcaa tggtgtcggt atcgaaggcc tgacgacaag gaaactcgct caaaagctgg
     3901 gagttgagca gcctaccctg tactggcacg tgaagaacaa gcgggccctg ctcgatgccc
     3961 tgccaatcga gatgctggac aggcatcata cccacttctg ccccctggaa ggcgagtcat
     4021 ggcaagactt tctgcggaac aacgccaagt cataccgctg tgctctcctc tcacatcgcg
     4081 acggggctaa agtgcatctc ggcacccgcc caacagagaa acagtacgaa accctggaaa
     4141 atcagctcgc gttcctgtgt cagcaaggct tctccctgga gaacgcactg tacgctctgt
     4201 ccgccgtggg ccactttaca ctgggctgcg tattggagga acaggagcat caagtagcaa
     4261 aagaggaaag agagacacct accaccgatt ctatgccccc acttctgaga caagcaattg
     4321 agctgttcga ccggcaggga gccgaacctg ccttcctttt cggcctggaa ctaatcatat
     4381 gtggcctgga gaaacagcta aagtgcgaaa gcggcgggcc gaccgacgcc cttgacgatt
     4441 ttgacttaga catgctccca gccgatgccc ttgacgactt tgaccttgat atgctgcctg
     4501 ctgacgctct tgacgatttt gaccttgaca tgctccccgg gtaactaagt aaggatcgat
     4561 ccaagatatc gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     4621 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     4681 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     4741 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     4801 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     4861 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     4921 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     4981 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     5041 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     5101 ctttgggccg cctccccgcc tgtttcgcct cggcgtccgg actagaggta cctttaagac
     5161 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     5221 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     5281 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     5341 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     5401 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     5461 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     5521 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5581 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5641 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc
     5701 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     5761 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     5821 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5881 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5941 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     6001 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     6061 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     6121 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     6181 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     6241 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     6301 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     6361 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6421 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6481 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6541 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6601 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6661 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6721 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6781 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6841 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6901 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6961 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7021 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     7081 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7141 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7201 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7261 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7321 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7381 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7441 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7501 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7561 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     7621 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     7681 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     7741 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     7801 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     7861 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     7921 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg
     7981 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
     8041 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
     8101 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
     8161 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
     8221 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     8281 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     8341 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     8401 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     8461 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     8521 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     8581 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     8641 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     8701 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     8761 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     8821 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     8881 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     8941 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     9001 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     9061 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     9121 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     9181 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     9241 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     9301 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     9361 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     9421 acaagcttaa tacactcctt aattg
//
