LOCUS       sequence-205398-       10577 bp    DNA     circular SYN 15-DEC-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 10577)
  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 10577)
  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: rLAMP1-mCherry-EGFP-Tet-On
FEATURES             Location/Qualifiers
     source          1..10577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      586..1174
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1075..1094
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1328..1345
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1462..1483
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1498..1503,1504..1521,1522..1528)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1536..1552
                     /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     1541..1563
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1560..1576
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1560..1576
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1595..1615
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1597..1615
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1643..1869
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1870..2050
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2097..2222
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2715..2948
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3133..3177
                     /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    3390..3507
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3561..3875
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    3569..3587
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3605..3623
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3640..3658
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3676..3694
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3712..3730
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3747..3765
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3783..3801
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3848..3872
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      3892..3901
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3896..3905
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3898..3960,3961..5118)
                     /codon_start=1
                     /product="rat lysosome-associated membrane glycoprotein 1"
                     /label=LAMP1
                     /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
                     LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
                     RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
                     VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
                     CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
                     MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
                     ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
                     SHAGYQTI"
     CDS             5143..5850
                     /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(5288..5306)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5556..5575
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    5878..5902
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      5911..5920
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5917..5919,5920..5922,5923..6636)
                     /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(5917..5919,5920..5922,5923..6633)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5962..5983)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6223..6242)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6570..6591
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        6664..7174
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     7093..7111
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             7184..7975
                     /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(7238..7257)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7848..7867
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             7985..8038
                     /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(8039..8656,8657..8662,8663..8698,8699..8701,8702..8737,
                     8738..8740,8741..8776,8777..8785)
                     /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(8825..8845)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9243..9254)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9459..9692
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9764..9885
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9801..9820)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9855..9874
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9925..10060
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        10060..10131
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10132..10200,10201..10577,1..415)
                     /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(10350..10369)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
       61 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
      121 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
      181 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
      241 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
      301 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
      361 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
      421 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
      481 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
      541 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
      601 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
      661 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
      721 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
      781 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
      841 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
      901 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
      961 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     1021 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     1081 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     1141 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     1201 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     1261 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     1321 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     1381 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     1441 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     1501 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     1561 aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca
     1621 aaagctggag ctgcaagctt aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg
     1681 gtaacgatga gttagcaaca tgccttacaa ggagagaaaa agcaccgtgc atgccgattg
     1741 gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc aacagacggg tctgacatgg
     1801 attggacgaa ccactgaatt gccgcattgc agagatattg tatttaagtg cctagctcga
     1861 tacataaacg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     1921 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     1981 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     2041 atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct
     2101 ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg
     2161 tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg
     2221 tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg
     2281 gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg
     2341 cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac
     2401 aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc
     2461 tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag
     2521 aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat cttcagacct
     2581 ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa
     2641 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa
     2701 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg
     2761 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag
     2821 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc
     2881 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa
     2941 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg
     3001 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag
     3061 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa
     3121 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg
     3181 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga
     3241 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag
     3301 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggaca attctcgacc
     3361 tcgagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac
     3421 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa
     3481 aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag agatccactt
     3541 tggccgcgaa tcgatatgtc gagtttactc cctatcagtg atagagaacg tatgtcgagt
     3601 ttactcccta tcagtgatag agaacgatgt cgagtttact ccctatcagt gatagagaac
     3661 gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgagttta ctccctatca
     3721 gtgatagaga acgtatgtcg agtttatccc tatcagtgat agagaacgta tgtcgagttt
     3781 actccctatc agtgatagag aacgtatgtc gaggtaggcg tgtacggtgg gaggcctata
     3841 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag aattggctag cgccaccatg
     3901 gcggccccgg gcgcccggcg gccgctgctc ctgttgctgc tggcaggcct tgcacacagc
     3961 gccccagcac tgttcgaggt gaaagacaac aacggcacag cgtgtataat ggccagcttc
     4021 tctgcctcct ttctgaccac ctatgaggct ggacatgttt ctaaggtctc gaatatgacc
     4081 ctgccagcct ctgcagaagt cctgaagaat agcagctctt gtggtgaaaa gaatgcttct
     4141 gagcccaccc tcgcaatcac ctttggagaa ggatatttac tgaaactcac cttcacaaaa
     4201 aacacaacac gttacagtgt ccagcacatg tatttcacat ataacctgtc agacacacaa
     4261 ttctttccca atgccagctc caaagggccc gacactgtgg attccacaac tgacatcaag
     4321 gcagacatca acaaaacata ccgatgtgtc agcgacatca gggtctacat gaagaatgtg
     4381 accattgtgc tctgggacgc tactatccag gcctacctgc cgagtagcaa cttcagcaag
     4441 gaagagacac gctgcccaca ggatcaacct tccccaacta ctgggccacc cagcccctca
     4501 ccaccacttg tgcccacaaa ccccagtgtg tccaagtaca atgtgactgg tgacaatgga
     4561 acctgcctgc tggcctctat ggcactgcaa ctcaacatca cctacatgaa gaaggacaac
     4621 acgactgtga ccagagcatt caacatcaac ccaagtgaca aatatagtgg gacttgcggt
     4681 gcccagttgg tgaccctgaa ggtggggaac aagagcagag tcctggagct gcagtttggg
     4741 atgaatgcca cttctagcct gtttttcctg caaggagttc agttgaacat gactcttcct
     4801 gatgccatag agcccacgtt cagcacctcc aactattccc tgaaagctct tcaggccagt
     4861 gtcggcaact catacaagtg caacagtgag gagcacatct ttgtcagcaa ggcgctcgcc
     4921 ctcaatgtct tcagcgtgca agtccaggct ttcagggtag aaagtgacag gtttgggtct
     4981 gtggaagagt gtgtacagga cggtaacaac atgctgatcc ccattgctgt gggcggggcc
     5041 ctggcagggc tggtcctcat cgtcctcatc gcctacctca tcggcaggaa gaggagtcac
     5101 gcgggctatc agaccatcgg tggatctgga ggttctggtg gaatggtgag caagggcgag
     5161 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
     5221 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     5281 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
     5341 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
     5401 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     5461 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     5521 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     5581 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
     5641 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
     5701 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
     5761 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     5821 tccaccggcg gcatggacga gctgtacaag tccggactca gatccgctgg cccaggaaca
     5881 agtttgtaca aaaaagcagg ctttccggtc gccaccatgg tgagcaaggg cgaggagctg
     5941 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     6001 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     6061 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     6121 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     6181 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     6241 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     6301 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     6361 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     6421 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     6481 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     6541 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     6601 gggatcactc tcggcatgga cgagctgtac aagtagtccg gactcagatc cggtaccgga
     6661 tccggggttg gggttgcgcc ttttccaagg cagccctggg tttgcgcagg gacgcggctg
     6721 ctctgggcgt ggttccggga aacgcagcgg cgccgaccct gggtctcgca cattcttcac
     6781 gtccgttcgc agcgtcaccc ggatcttcgc cgctaccctt gtgggccccc cggcgacgct
     6841 tcctgctccg cccctaagtc gggaaggttc cttgcggttc gcggcgtgcc ggacgtgaca
     6901 aacggaagcc gcacgtctca ctagtaccct cgcagacgga cagcgccagg gagcaatggc
     6961 agcgcgccga ccgcgatggg ctgtggccaa tagcggctgc tcagcagggc gcgccgagag
     7021 cagcggccgg gaaggggcgg tgcgggaggc ggggtgtggg gcggtagtgt gggccctgtt
     7081 cctgcccgcg cggtgttccg cattctgcaa gcctccggag cgcacgtcgg cagtcggctc
     7141 cctcgttgac cgaatcaccg acctctctcc ccagcaattc accatgattg aacaagatgg
     7201 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     7261 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     7321 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     7381 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     7441 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     7501 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     7561 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     7621 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     7681 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     7741 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     7801 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     7861 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     7921 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttccccgg
     7981 tgccgaaggt agaggttctc tcctcacttg tggtgatgtt gaagaaaacc ctggtccaat
     8041 gtctagactg gacaagagca aagtcataaa cggagctctg gaattactca atggtgtcgg
     8101 tatcgaaggc ctgacgacaa ggaaactcgc tcaaaagctg ggagttgagc agcctaccct
     8161 gtactggcac gtgaagaaca agcgggccct gctcgatgcc ctgccaatcg agatgctgga
     8221 caggcatcat acccacttct gccccctgga aggcgagtca tggcaagact ttctgcggaa
     8281 caacgccaag tcataccgct gtgctctcct ctcacatcgc gacggggcta aagtgcatct
     8341 cggcacccgc ccaacagaga aacagtacga aaccctggaa aatcagctcg cgttcctgtg
     8401 tcagcaaggc ttctccctgg agaacgcact gtacgctctg tccgccgtgg gccactttac
     8461 actgggctgc gtattggagg aacaggagca tcaagtagca aaagaggaaa gagagacacc
     8521 taccaccgat tctatgcccc cacttctgag acaagcaatt gagctgttcg accggcaggg
     8581 agccgaacct gccttccttt tcggcctgga actaatcata tgtggcctgg agaaacagct
     8641 aaagtgcgaa agcggcgggc cgaccgacgc ccttgacgat tttgacttag acatgctccc
     8701 agccgatgcc cttgacgact ttgaccttga tatgctgcct gctgacgctc ttgacgattt
     8761 tgaccttgac atgctccccg ggtaactaag taaggatcga tccaagatat cgtattctta
     8821 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     8881 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     8941 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     9001 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     9061 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     9121 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     9181 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     9241 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     9301 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     9361 ctgtttcgcc tcggcgtccg gactagaggt acctttaaga ccaatgactt acaaggcagc
     9421 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     9481 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     9541 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     9601 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     9661 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     9721 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
     9781 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     9841 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
     9901 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     9961 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
    10021 gagctattcc agaagtagtg aggaggcttt tttggaggcc tttcaaatat gtatccgctc
    10081 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
    10141 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
    10201 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
    10261 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
    10321 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
    10381 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
    10441 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
    10501 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
    10561 aaggagctaa ccgcttt
//
