LOCUS       sequence-213506-       10576 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Tet-On construct to induce expression of tandem eGFP-mCherry tagged 
            mLAMP1 (lysosome marker).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10576)
  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 10576)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: rLAMP1-mCherry-EGFP-Tet-On
FEATURES             Location/Qualifiers
     source          1..10576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        54..368
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    62..80
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    98..116
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    133..151
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    169..187
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    205..223
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    240..258
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    276..294
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     341..365
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      385..394
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      389..398
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(391..453,454..1611)
                     /codon_start=1
                     /product="rat lysosome-associated membrane glycoprotein 1"
                     /label=LAMP1
                     /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
                     LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
                     RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
                     VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
                     CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
                     MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
                     ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
                     SHAGYQTI"
     CDS             1636..2343
                     /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(1781..1799)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2049..2068
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    2371..2395
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2404..2413
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2410..2412,2413..2415,2416..3129)
                     /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(2410..2412,2413..2415,2416..3126)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2455..2476)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2716..2735)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3063..3084
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        3157..3666
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     3586..3604
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             3676..4467
                     /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(3730..3749)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4340..4359
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             4477..4530
                     /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(4531..5148,5149..5154,5155..5190,5191..5193,5194..5229,
                     5230..5232,5233..5268,5269..5277)
                     /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(5317..5337)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5735..5746)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5951..6184
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6256..6377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6293..6312)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6347..6366
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6417..6552
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        6552..6623
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6624..6692,6693..7484)
                     /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(6842..6861)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7655..8243
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8144..8163
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8397..8414
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8531..8552
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8567..8572,8573..8590,8591..8597)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8605..8621
                     /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     8610..8632
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8629..8645
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8629..8645
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8664..8684
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8666..8684
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        8712..8938
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8939..9119
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9166..9291
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9784..10017
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10202..10246
                     /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    10459..10576
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cgggtttatt acagggacag cagagatcca ctttggccgc gaatcgatat gtcgagttta
       61 ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacga
      121 tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg
      181 atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttat
      241 ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat
      301 gtcgaggtag gcgtgtacgg tgggaggcct atataagcag agctcgttta gtgaaccgtc
      361 agatcgcctg gagaattggc tagcgccacc atggcggccc cgggcgcccg gcggccgctg
      421 ctcctgttgc tgctggcagg ccttgcacac agcgccccag cactgttcga ggtgaaagac
      481 aacaacggca cagcgtgtat aatggccagc ttctctgcct cctttctgac cacctatgag
      541 gctggacatg tttctaaggt ctcgaatatg accctgccag cctctgcaga agtcctgaag
      601 aatagcagct cttgtggtga aaagaatgct tctgagccca ccctcgcaat cacctttgga
      661 gaaggatatt tactgaaact caccttcaca aaaaacacaa cacgttacag tgtccagcac
      721 atgtatttca catataacct gtcagacaca caattctttc ccaatgccag ctccaaaggg
      781 cccgacactg tggattccac aactgacatc aaggcagaca tcaacaaaac ataccgatgt
      841 gtcagcgaca tcagggtcta catgaagaat gtgaccattg tgctctggga cgctactatc
      901 caggcctacc tgccgagtag caacttcagc aaggaagaga cacgctgccc acaggatcaa
      961 ccttccccaa ctactgggcc acccagcccc tcaccaccac ttgtgcccac aaaccccagt
     1021 gtgtccaagt acaatgtgac tggtgacaat ggaacctgcc tgctggcctc tatggcactg
     1081 caactcaaca tcacctacat gaagaaggac aacacgactg tgaccagagc attcaacatc
     1141 aacccaagtg acaaatatag tgggacttgc ggtgcccagt tggtgaccct gaaggtgggg
     1201 aacaagagca gagtcctgga gctgcagttt gggatgaatg ccacttctag cctgtttttc
     1261 ctgcaaggag ttcagttgaa catgactctt cctgatgcca tagagcccac gttcagcacc
     1321 tccaactatt ccctgaaagc tcttcaggcc agtgtcggca actcatacaa gtgcaacagt
     1381 gaggagcaca tctttgtcag caaggcgctc gccctcaatg tcttcagcgt gcaagtccag
     1441 gctttcaggg tagaaagtga caggtttggg tctgtggaag agtgtgtaca ggacggtaac
     1501 aacatgctga tccccattgc tgtgggcggg gccctggcag ggctggtcct catcgtcctc
     1561 atcgcctacc tcatcggcag gaagaggagt cacgcgggct atcagaccat cggtggatct
     1621 ggaggttctg gtggaatggt gagcaagggc gaggaggata acatggccat catcaaggag
     1681 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag
     1741 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     1801 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
     1861 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc
     1921 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
     1981 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc
     2041 tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg
     2101 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc
     2161 ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     2221 ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc
     2281 gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac
     2341 aagtccggac tcagatccgc tggcccagga acaagtttgt acaaaaaagc aggctttccg
     2401 gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     2461 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     2521 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     2581 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     2641 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     2701 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     2761 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     2821 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     2881 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     2941 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     3001 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     3061 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     3121 tacaagtagt ccggactcag atccggtacc ggatccgggg ttggggttgc gccttttcca
     3181 aggcagccct gggtttgcgc agggacgcgg ctgctctggg cgtggttccg ggaaacgcag
     3241 cggcgccgac cctgggtctc gcacattctt cacgtccgtt cgcagcgtca cccggatctt
     3301 cgccgctacc cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg
     3361 ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc tcactagtac
     3421 cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc
     3481 caatagcggc tgctcagcag ggcgcgccga gagcagcggc cgggaagggg cggtgcggga
     3541 ggcggggtgt ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg
     3601 caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctt
     3661 ccccagcaat tcaccatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3721 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3781 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3841 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3901 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3961 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4021 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4081 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4141 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4201 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4261 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4321 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4381 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4441 ttctatcgcc ttcttgacga gttcttcccc ggtgccgaag gtagaggttc tctcctcact
     4501 tgtggtgatg ttgaagaaaa ccctggtcca atgtctagac tggacaagag caaagtcata
     4561 aacggagctc tggaattact caatggtgtc ggtatcgaag gcctgacgac aaggaaactc
     4621 gctcaaaagc tgggagttga gcagcctacc ctgtactggc acgtgaagaa caagcgggcc
     4681 ctgctcgatg ccctgccaat cgagatgctg gacaggcatc atacccactt ctgccccctg
     4741 gaaggcgagt catggcaaga ctttctgcgg aacaacgcca agtcataccg ctgtgctctc
     4801 ctctcacatc gcgacggggc taaagtgcat ctcggcaccc gcccaacaga gaaacagtac
     4861 gaaaccctgg aaaatcagct cgcgttcctg tgtcagcaag gcttctccct ggagaacgca
     4921 ctgtacgctc tgtccgccgt gggccacttt acactgggct gcgtattgga ggaacaggag
     4981 catcaagtag caaaagagga aagagagaca cctaccaccg attctatgcc cccacttctg
     5041 agacaagcaa ttgagctgtt cgaccggcag ggagccgaac ctgccttcct tttcggcctg
     5101 gaactaatca tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg gccgaccgac
     5161 gcccttgacg attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt
     5221 gatatgctgc ctgctgacgc tcttgacgat tttgaccttg acatgctccc cgggtaacta
     5281 agtaaggatc gatccaagat atcgtattct taactatgtt gctcctttta cgctatgtgg
     5341 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     5401 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     5461 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     5521 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     5581 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     5641 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     5701 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     5761 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     5821 gagtcggatc tccctttggg ccgcctcccc gcctgtttcg cctcggcgtc cggactagag
     5881 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     5941 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     6001 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     6061 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     6121 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     6181 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     6241 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6301 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6361 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc
     6421 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6481 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     6541 tttttggagg cctttcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     6601 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     6661 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     6721 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     6781 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     6841 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     6901 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6961 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     7021 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     7081 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     7141 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     7201 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     7261 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     7321 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     7381 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     7441 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     7501 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     7561 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     7621 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     7681 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     7741 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     7801 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     7861 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7921 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7981 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     8041 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     8101 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     8161 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     8221 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     8281 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     8341 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     8401 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     8461 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     8521 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     8581 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     8641 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
     8701 tgcaagctta atgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag
     8761 ttagcaacat gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag
     8821 gtggtacgat cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac
     8881 cactgaattg ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg
     8941 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     9001 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     9061 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     9121 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
     9181 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
     9241 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
     9301 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
     9361 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
     9421 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
     9481 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
     9541 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
     9601 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga
     9661 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     9721 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     9781 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     9841 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     9901 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     9961 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
    10021 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
    10081 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
    10141 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
    10201 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
    10261 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
    10321 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
    10381 attatcgttt cagacccacc tcccaacccc gaggggacaa ttctcgacct cgagacaaat
    10441 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
    10501 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
    10561 aaaaattcaa aatttt
//
