LOCUS       sequence-205399-        9446 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 9446)
  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 9446)
  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..9446
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    41..158
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        212..526
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    220..238
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    256..274
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    291..309
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    327..345
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    363..381
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    398..416
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    434..452
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     499..523
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      543..552
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             663..1370
                     /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(808..826)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1076..1095
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    1398..1422
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1431..1440
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1437..1439,1440..1442,1443..2156)
                     /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(1437..1439,1440..1442,1443..2153)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1482..1503)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1743..1762)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2090..2111
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        2184..2694
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2613..2631
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2704..3495
                     /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(2758..2777)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3368..3387
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             3505..3558
                     /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(3559..4176,4177..4182,4183..4218,4219..4221,4222..4257,
                     4258..4260,4261..4296,4297..4305)
                     /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(4345..4365)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4763..4774)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4979..5212
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5284..5405
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5321..5340)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5375..5394
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5445..5580
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5580..5651
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5652..5720,5721..6512)
                     /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(5870..5889)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6683..7271
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7172..7191
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7425..7442
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7559..7580
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7595..7600,7601..7618,7619..7625)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7633..7649
                     /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     7638..7660
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7657..7673
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7657..7673
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7692..7712
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7694..7712
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7740..7966
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7967..8147
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8194..8319
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8812..9045
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9230..9274
                     /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"
ORIGIN
        1 aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
       61 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
      121 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
      181 gagatccact ttggccgcga atcgatatgt cgagtttact ccctatcagt gatagagaac
      241 gtatgtcgag tttactccct atcagtgata gagaacgatg tcgagtttac tccctatcag
      301 tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt
      361 actccctatc agtgatagag aacgtatgtc gagtttatcc ctatcagtga tagagaacgt
      421 atgtcgagtt tactccctat cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg
      481 ggaggcctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gaattggcta
      541 gcgccaccat ggtgggtcgg aacagcgcca tcgccgccgg tgtatgcggg gcccttttca
      601 ttgggtactg catctacttc gaccgcaaaa gacgaagtgg tggatctgga ggttctggtg
      661 gaatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
      721 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
      781 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
      841 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
      901 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
      961 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     1021 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     1081 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     1141 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     1201 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     1261 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     1321 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag tccggactca
     1381 gatccgctgg cccaggaaca agtttgtaca aaaaagcagg ctttccggtc gccaccatgg
     1441 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1501 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1561 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1621 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     1681 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1741 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1801 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1861 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     1921 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     1981 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2041 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2101 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtagtccg
     2161 gactcagatc cggtaccgga tccggggttg gggttgcgcc ttttccaagg cagccctggg
     2221 tttgcgcagg gacgcggctg ctctgggcgt ggttccggga aacgcagcgg cgccgaccct
     2281 gggtctcgca cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc cgctaccctt
     2341 gtgggccccc cggcgacgct tcctgctccg cccctaagtc gggaaggttc cttgcggttc
     2401 gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca ctagtaccct cgcagacgga
     2461 cagcgccagg gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa tagcggctgc
     2521 tcagcagggc gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc ggggtgtggg
     2581 gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg cattctgcaa gcctccggag
     2641 cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg acctctctcc ccagcaattc
     2701 accatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     2761 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     2821 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     2881 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     2941 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3001 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3061 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3121 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3181 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     3241 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3301 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3361 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     3421 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     3481 cttgacgagt tcttccccgg tgccgaaggt agaggttctc tcctcacttg tggtgatgtt
     3541 gaagaaaacc ctggtccaat gtctagactg gacaagagca aagtcataaa cggagctctg
     3601 gaattactca atggtgtcgg tatcgaaggc ctgacgacaa ggaaactcgc tcaaaagctg
     3661 ggagttgagc agcctaccct gtactggcac gtgaagaaca agcgggccct gctcgatgcc
     3721 ctgccaatcg agatgctgga caggcatcat acccacttct gccccctgga aggcgagtca
     3781 tggcaagact ttctgcggaa caacgccaag tcataccgct gtgctctcct ctcacatcgc
     3841 gacggggcta aagtgcatct cggcacccgc ccaacagaga aacagtacga aaccctggaa
     3901 aatcagctcg cgttcctgtg tcagcaaggc ttctccctgg agaacgcact gtacgctctg
     3961 tccgccgtgg gccactttac actgggctgc gtattggagg aacaggagca tcaagtagca
     4021 aaagaggaaa gagagacacc taccaccgat tctatgcccc cacttctgag acaagcaatt
     4081 gagctgttcg accggcaggg agccgaacct gccttccttt tcggcctgga actaatcata
     4141 tgtggcctgg agaaacagct aaagtgcgaa agcggcgggc cgaccgacgc ccttgacgat
     4201 tttgacttag acatgctccc agccgatgcc cttgacgact ttgaccttga tatgctgcct
     4261 gctgacgctc ttgacgattt tgaccttgac atgctccccg ggtaactaag taaggatcga
     4321 tccaagatat cgtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     4381 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     4441 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     4501 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     4561 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     4621 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     4681 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     4741 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     4801 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     4861 cctttgggcc gcctccccgc ctgtttcgcc tcggcgtccg gactagaggt acctttaaga
     4921 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     4981 gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc
     5041 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     5101 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     5161 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt
     5221 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
     5281 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5341 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5401 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc
     5461 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5521 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5581 tttcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5641 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5701 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5761 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5821 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5881 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5941 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6001 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     6061 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     6121 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6181 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     6241 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     6301 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6361 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6421 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6481 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6541 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6601 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6661 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6721 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6781 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6841 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6901 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6961 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7021 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7081 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7141 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7201 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7261 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7321 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     7381 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     7441 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     7501 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     7561 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     7621 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     7681 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat
     7741 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc
     7801 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg
     7861 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc
     7921 gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt ctctctggtt
     7981 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     8041 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     8101 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac
     8161 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct
     8221 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact
     8281 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt
     8341 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa
     8401 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa
     8461 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca
     8521 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata
     8581 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag
     8641 accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa
     8701 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
     8761 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
     8821 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
     8881 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
     8941 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
     9001 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
     9061 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
     9121 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
     9181 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
     9241 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
     9301 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
     9361 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
     9421 gacccacctc ccaaccccga ggggac
//
