LOCUS       sequence-205400-       10166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-On construct to induce expression of tandem mCherry-eGFP tagged 
            PMP34 (peroxisome marker).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10166)
  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 10166)
  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: PMP34-mCherry-EGFP-Tet-On
FEATURES             Location/Qualifiers
     source          1..10166
                     /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"
     CDS             1383..2090
                     /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(1528..1546)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1796..1815
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    2118..2142
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2151..2160
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2157..2159,2160..2162,2163..2876)
                     /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(2157..2159,2160..2162,2163..2873)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2202..2223)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2463..2482)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2810..2831
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        2904..3414
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3333..3351
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3424..4215
                     /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(3478..3497)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4088..4107
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             4225..4278
                     /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(4279..4896,4897..4902,4903..4938,4939..4941,4942..4977,
                     4978..4980,4981..5016,5017..5025)
                     /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(5065..5085)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5483..5494)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5699..5932
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6004..6125
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6041..6060)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6095..6114
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6165..6300
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6300..6371
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6372..6440,6441..7232)
                     /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(6590..6609)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7403..7991
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7892..7911
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8145..8162
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8279..8300
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8315..8320,8321..8338,8339..8345)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8353..8369
                     /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     8358..8380
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8377..8393
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8377..8393
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8412..8432
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8414..8432
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8460..8686
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8687..8867
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8914..9039
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9532..9765
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9950..9994
                     /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 gattgatgag aaaagaaaat ccaaaactac acacatggtg ctcctggaga
      601 tcattaaaga agaaggactc ctggcaccat atcgagggtg gtttccagtg atttccagtc
      661 tctgctgctc caattttgtc tatttctaca cttttaatag cctcaaagca ctctgggtca
      721 aaggtcaaca ttctaccact ggaaaagatc tggtagttgg gtttgttgca ggagtggtta
      781 atgtgttgct aacaactcca ctctgggtgg taaacaccag actgaagctt caaggagcaa
      841 aatttaggaa tgaagacatt gtaccaacaa actacaaagg tatcattgat gcttttcatc
      901 agatcattcg cgatgaagga atctcggctt tatggaatgg cacatttccc tcattgctgt
      961 tggtcttcaa tcctgccatc cagttcatgt tttatgaagg tttaaaacgg cagcttttaa
     1021 agaaacggat gaagctttct tccttggatg tgttcatcat tggtgcagta gccaaagcga
     1081 ttgccaccac ggtgacctat cccctgcaga cggtacagtc aattctgagg tttgggcgtc
     1141 atagactaaa cccagaaaac agaacattgg gaagtcttcg gaatattctc tatcttcttc
     1201 accaacgagt aagacgtttt ggaataatgg gactctacaa aggccttgaa gccaaactgc
     1261 tgcagacagt cctcactgct gctctcatgt tccttgttta tgagaaactg acagctgcca
     1321 ccttcacagt tatggggctg aagcgtgcac accaacacgg tggatctgga ggttctggtg
     1381 gaatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     1441 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     1501 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     1561 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     1621 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     1681 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     1741 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     1801 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     1861 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     1921 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     1981 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     2041 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag tccggactca
     2101 gatccgctgg cccaggaaca agtttgtaca aaaaagcagg ctttccggtc gccaccatgg
     2161 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     2221 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     2281 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     2341 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     2401 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     2461 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     2521 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     2581 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     2641 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     2701 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2761 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2821 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtagtccg
     2881 gactcagatc cggtaccgga tccggggttg gggttgcgcc ttttccaagg cagccctggg
     2941 tttgcgcagg gacgcggctg ctctgggcgt ggttccggga aacgcagcgg cgccgaccct
     3001 gggtctcgca cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc cgctaccctt
     3061 gtgggccccc cggcgacgct tcctgctccg cccctaagtc gggaaggttc cttgcggttc
     3121 gcggcgtgcc ggacgtgaca aacggaagcc gcacgtctca ctagtaccct cgcagacgga
     3181 cagcgccagg gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa tagcggctgc
     3241 tcagcagggc gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc ggggtgtggg
     3301 gcggtagtgt gggccctgtt cctgcccgcg cggtgttccg cattctgcaa gcctccggag
     3361 cgcacgtcgg cagtcggctc cctcgttgac cgaatcaccg acctctctcc ccagcaattc
     3421 accatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3481 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3541 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3601 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3661 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3721 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3781 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3841 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3901 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     3961 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4021 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4081 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4141 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4201 cttgacgagt tcttccccgg tgccgaaggt agaggttctc tcctcacttg tggtgatgtt
     4261 gaagaaaacc ctggtccaat gtctagactg gacaagagca aagtcataaa cggagctctg
     4321 gaattactca atggtgtcgg tatcgaaggc ctgacgacaa ggaaactcgc tcaaaagctg
     4381 ggagttgagc agcctaccct gtactggcac gtgaagaaca agcgggccct gctcgatgcc
     4441 ctgccaatcg agatgctgga caggcatcat acccacttct gccccctgga aggcgagtca
     4501 tggcaagact ttctgcggaa caacgccaag tcataccgct gtgctctcct ctcacatcgc
     4561 gacggggcta aagtgcatct cggcacccgc ccaacagaga aacagtacga aaccctggaa
     4621 aatcagctcg cgttcctgtg tcagcaaggc ttctccctgg agaacgcact gtacgctctg
     4681 tccgccgtgg gccactttac actgggctgc gtattggagg aacaggagca tcaagtagca
     4741 aaagaggaaa gagagacacc taccaccgat tctatgcccc cacttctgag acaagcaatt
     4801 gagctgttcg accggcaggg agccgaacct gccttccttt tcggcctgga actaatcata
     4861 tgtggcctgg agaaacagct aaagtgcgaa agcggcgggc cgaccgacgc ccttgacgat
     4921 tttgacttag acatgctccc agccgatgcc cttgacgact ttgaccttga tatgctgcct
     4981 gctgacgctc ttgacgattt tgaccttgac atgctccccg ggtaactaag taaggatcga
     5041 tccaagatat cgtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     5101 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5161 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     5221 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     5281 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     5341 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     5401 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     5461 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     5521 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     5581 cctttgggcc gcctccccgc ctgtttcgcc tcggcgtccg gactagaggt acctttaaga
     5641 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     5701 gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc
     5761 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     5821 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     5881 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt
     5941 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
     6001 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     6061 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     6121 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc
     6181 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     6241 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     6301 tttcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6361 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     6421 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     6481 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     6541 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     6601 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     6661 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6721 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     6781 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     6841 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6901 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     6961 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     7021 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     7081 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     7141 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     7201 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     7261 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     7321 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     7381 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     7441 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     7501 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     7561 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7621 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7681 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7741 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7801 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7861 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7921 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7981 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     8041 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     8101 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     8161 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     8221 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     8281 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     8341 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     8401 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat
     8461 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc
     8521 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg
     8581 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc
     8641 gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt ctctctggtt
     8701 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     8761 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     8821 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac
     8881 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct
     8941 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact
     9001 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt
     9061 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa
     9121 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa
     9181 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca
     9241 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata
     9301 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag
     9361 accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa
     9421 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
     9481 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
     9541 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
     9601 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
     9661 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
     9721 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
     9781 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
     9841 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
     9901 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
     9961 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
    10021 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
    10081 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
    10141 gacccacctc ccaaccccga ggggac
//
