LOCUS       sequence-432520-       11986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable, inducible expression in lysosome.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11986)
  AUTHORS   Zhong X, Li Q, Polacco BJ, Patil T, DiBerto JF, Vartak R, Xu J, 
            Marley A, Foussard H, Roth BL, Eckhardt M, Von Zastrow M, Krogan NJ,
            Huttenhain R
  TITLE     An automated proximity proteomics pipeline for subcellular proteome 
            and protein interaction mapping.
  JOURNAL   bioRxiv. 2023 Apr 12:2023.04.11.536358. doi: 
            10.1101/2023.04.11.536358. Preprint.
  PUBMED    37090610
REFERENCE   2  (bases 1 to 11986)
  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: "bioRxiv. 
            2023 Apr 12:2023.04.11.536358. doi: 10.1101/2023.04.11.536358. 
            Preprint."
COMMENT     Sequence Label: LAMP1-APEX2
FEATURES             Location/Qualifiers
     source          1..11986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             732..758
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     polyA_signal    945..1079
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    1123..1238
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     CDS             complement(1501..1545)
                     /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    1730..1963
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2460..2585
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             2632..3265
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    3433..3567
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        3615..3719
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3720..3788,3789..4580)
                     /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(3938..3957)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4751..5339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5240..5259
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5493..5510
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5627..5648
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5663..5668,5669..5686,5687..5693)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5701..5717
                     /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     5706..5728
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5725..5741
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5725..5741
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     LTR             5869..6502
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    6709..7297
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6815..6826
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7224..7244
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7311..7910)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7394..7414)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7891..7910
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(7919..8248)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7933..8068
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             complement(8259..9005)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     promoter        complement(9024..9534)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(9087..9105)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     promoter        9551..9918
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    9559..9577
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9595..9613
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9631..9649
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9667..9685
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9703..9721
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9739..9757
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9775..9793
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      9926..9935
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(9928..10011,10012..11178)
                     /codon_start=1
                     /product="human lysosome-associated membrane glycoprotein 
                     1"
                     /label=LAMP1
                     /translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN
                     FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF
                     TRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM
                     NNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY
                     NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE
                     GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA
                     EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIV
                     LIAYLVGRKRSHAGYQTI"
     CDS             join(11203..11205,11206..11208,11209..11919)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(11203..11205,11206..11208,11209..11919)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(11248..11269)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(11509..11528)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     11856..11877
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      11932..11941
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             11941..11964
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(11965..11986,1..725)
                     /codon_start=1
                     /product="soybean ascorbate peroxidase, engineered to serve
                     as a monomeric tag for electron microscopy and live-cell 
                     proteomics (Lam et al., 2015)"
                     /label=APEX2
                     /note="more active than the original APEX due to the A134P 
                     mutation"
                     /translation="GKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAGT
                     FDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVAV
                     EVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGHT
                     IGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVDK
                     YAADEDAFFADYAEAHQKLSELGFADA"
ORIGIN
        1 gtgctgatta ccaggacgcc gttgagaagg cgaagaagaa gctcagaggc ttcatcgctg
       61 agaagagatg cgctcctcta atgctccgtt tggcattcca ctctgctgga acctttgaca
      121 agggcacgaa gaccggtgga cccttcggaa ccatcaagca ccctgccgaa ctggctcaca
      181 gcgctaacaa cggtcttgac atcgctgtta ggcttttgga gccactcaag gcggagttcc
      241 ctattttgag ctacgccgat ttctaccagt tggctggcgt tgttgccgtt gaggtcacgg
      301 gtggacctaa ggttccattc caccctggaa gagaggacaa gcctgagcca ccaccagagg
      361 gtcgcttgcc cgatcccact aagggttctg accatttgag agatgtgttt ggcaaagcta
      421 tggggcttac tgaccaagat atcgttgctc tatctggggg tcacactatt ggagctgcac
      481 acaaggagcg ttctggattt gagggtccct ggacctctaa tcctcttatt ttcgacaact
      541 catacttcac ggagttgttg agtggtgaga aggaaggtct ccttcagcta ccttctgaca
      601 aggctctttt gtctgaccct gtattccgcc ctctcgttga caaatatgca gcggacgaag
      661 atgccttctt tgctgattac gctgaggctc accaaaagct ttccgagctt gggtttgctg
      721 atgccctgca gctgcctccc ctggagcgcc tgaccctgga ctaaggatcc agaccacctc
      781 ccctgcgagc taagctggac agccaatgac gggtaagaga gtgacatttt tcactaacct
      841 aagacaggag ggccgtcaga gctactgcct aatccaaaga cgggtaaaag tgataaaaat
      901 gtatcactcc aacctaagac aggcgcagct tccgagggat ttgagatcca gacatgataa
      961 gatacattga tgagtttgga caaaccaaaa ctagaatgca gtgaaaaaaa tgccttattt
     1021 gtgaaatttg tgatgctatt gccttatttg taaccattat aagctgcaat aaacaagtta
     1081 agtcgataaa ctggatctct gctgtccctg taataaaccc gaaaattttg aatttttgta
     1141 atttgttttt gtaattcttt agtttgtatg tctgttgcta ttatgtctac tattctttcc
     1201 cctgcactgt accccccaat cccccctttt cttttaaagg cgataccgtc gagatccgtt
     1261 cactaatcga atggatctgt ctctgtctct ctctccacct tcttcttcta ttccttcggg
     1321 cctgtcgggt cccctcgggg ttgggaggtg ggtctgaaac gataatggtg aatatccctg
     1381 cctaactcta ttcactatag aaagtacagc aaaaactatt cttaaaccta ccaagcctcc
     1441 tactatcatt atgaataatt ttatatacca cagccaattt gttatgttaa accaattcca
     1501 caaacttgcc catttatcta attccaataa ttcttgttca ttcttttctt gctggttttg
     1561 cgattcttca attaaggagt gtattaagct tgtgtaattg ttaatttctc tgtcccactc
     1621 catccaggtc gtgtgattcc aaatctgttc cagagattta ttactccaac tagcattcca
     1681 aggcacagca gtggtgcaaa tgagttttcc agagcaaccc caaatcccca ggagctgttg
     1741 atcctttagg tatctttcca cagccaggat tcttgcctgg agctgcttga tgccccagac
     1801 tgtgagttgc aacagatgct gttgcgcctc aatagccctc agcaaattgt tctgctgctg
     1861 cactatacca gacaataatt gtctggcctg taccgtcagc gtcattgacg ctgcgcccat
     1921 agtgcttcct gctgctccca agaacccaag gaacaaagct cctattccca ctgctctttt
     1981 ttctctctgc accactcttc tctttgcctt ggtgggtgct actcctaatg gttcaatttt
     2041 tactacttta tatttatata attcacttct ccaattgtcc ctcatatctc ctcctccagg
     2101 tctgaagatc agcggccggc cgcttgctgt gcggtggtct tacttttgtt ttgctcttcc
     2161 tctatcttgt ctaaagcttc cttggtgtct tttatctcta tcctttgatg cacacaatag
     2221 agggttgcta ctgtattata taatgatcta agttcttctg atcctgtctg aagggatggt
     2281 tgtagctgtc ccagtatttg tctacagcct tctgatgttt ctaacaggcc aggattaact
     2341 gcgaatcgtt ctagctccct gcttgcccat actatatgtt ttaatttata ttttttcttt
     2401 ccccctggcc ttaaccgaat tttttcccat cgcgatctaa ttctcccccg cttaatactg
     2461 acgctctcgc acccatctct ctccttctag cctccgctag tcaaaatttt tggcgtactc
     2521 accagtcgcc gcccctcgcc tcttgccgtg cgcgcttcag caagccgagt cctgcgtcga
     2581 gagagctcct ctggtttccc tttcgctttc aagtccctgt tcgggcgcca ctgctagaga
     2641 ttttccacac tgactaaaag ggtctgaggg atctctagtt accagagtca cacaacagac
     2701 gggcacacac tacttgaagc actcaaggca agctttattg aggcttaagc agtgggttcc
     2761 ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac ccagtacagg
     2821 caaaaagcag ctgcttatat gcaggatctg agggctcgcc actccccagt cccgcccagg
     2881 ccacgcctcc ctggaaagtc cccagcggaa agtcccttgt agcaagctcg atatcagcag
     2941 ttcttgaagt actccggatg cagctctcgg gccacgtgat gaaatgctag gcggctgtca
     3001 aacctccact ctaacacttc tctctccggg tcatccatcc catgcaggct cacagggtgt
     3061 aacaagctgg tgttctctcc tttattggcc tcttctacct tatctggctc aactggtact
     3121 agcttgtagc accatccaaa ggtcagtgga tatctgaccc ctggccctgg tgtgtagttc
     3181 tgctaatcag ggaagtagcc ttgtgtgtgg tagatccaca gatcaaggat atcttgtctt
     3241 ctttgggagt gaattagccc ttccaactac taagtttgta gtacatattt aacaaataca
     3301 atttctttaa aatgaaaata attcagagga atcacaggtt tagagtaaat gaaaccacag
     3361 gtaattggca gtggtaatag ggtatggggt gggaagtttg ggatgatttt ggttagcttg
     3421 agttatccag ttgatccaga catgataaga tacattgatg agtttggaca aaccacaact
     3481 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     3541 accattataa gctgcaataa acaagttgat ctcccgatcc gtcgacgtca ggtggcactt
     3601 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3661 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3721 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3781 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3841 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3901 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3961 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4021 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4081 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4141 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4201 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4261 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4321 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4381 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4441 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4501 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4561 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4621 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4681 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4741 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4801 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4861 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4921 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4981 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5041 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5101 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5161 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5221 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5281 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5341 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5401 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     5461 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     5521 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     5581 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     5641 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     5701 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
     5761 agctagaggt cgacggtaaa cgctagcaat gtcaaggcct ctcactctct gatattcatt
     5821 tctttgcaag ttataaatac tgaataataa gatgacatga actactactg ctagagattt
     5881 tccacactga ctaaaagggt ctgagggatc tctagttacc agagtcacac aacagacggg
     5941 cacacactac ttgaagcact caaggcaagc tttattgagg cttaagcagt gggttcccta
     6001 gttagccaga gagctcccag gctcagatct ggtctaacca gagagaccca gtacaggcaa
     6061 aaagcagctg cttatatgca ggatctgagg gctcgccact ccccagtccc gcccaggcca
     6121 cgcctccctg gaaagtcccc agcggaaagt cccttgtagc aagctcgata tcagcagttc
     6181 ttgaagtact ccggatgcag ctctcgggcc acgtgatgaa atgctaggcg gctgtcaaac
     6241 ctccactcta acacttctct ctccgggtca tccatcccat gcaggctcac agggtgtaac
     6301 aagctggtgt tctctccttt attggcctct tctaccttat ctggctcaac tggtactagc
     6361 ttgtagcacc atccaaaggt cagtggatat ctgacccctg gccctggtgt gtagttctgc
     6421 taatcaggga agtagccttg tgtgtggtag atccacagat caaggatatc ttgtcttcgt
     6481 tgggagtgaa ttagcccttc cagtcccctc ttttctttta aaaagtggct aagatctaca
     6541 gctgccttgt aagtcattgg tcttaaaggt acctgaggtg tgactggaaa acccacctcc
     6601 tcctcctctt gtgcttctag ccaggcacaa tcagcattgg tagctgctgt attgctactt
     6661 gtgattgctc catgtttttc taggtctcga ctgcagaatt aattccaggc ggggaggcgg
     6721 cccaaaggga gatccgactc gtctgagggc gaaggcgaag acgcggaaga ggccgcagag
     6781 ccggcagcag gccgcgggaa ggaaggtccg ctggattgag ggccgaaggg acgtagcaga
     6841 aggacgtccc gcgcagaatc caggtggcaa cacaggcgag cagccatgga aaggacgtca
     6901 gcttccccga caacaccacg gaattgtcag tgcccaacag ccgagcccct gtccagcagc
     6961 gggcaaggca ggcggcgatg agttccgccg tggcaatagg gagggggaaa gcgaaagtcc
     7021 cggaaaggag ctgacaggtg gtggcaatgc cccaaccagt gggggttgcg tcagcaaaca
     7081 cagtgcacac cacgccacgt tgcctgacaa cgggccacaa ctcctcataa agagacagca
     7141 accaggattt atacaaggag gagaaaatga aagccatacg ggaagcaata gcatgataca
     7201 aaggcattaa agcagcgtat ccacatagcg taaaaggagc aacatagtta agaataccag
     7261 tcaatctttc acaaattttg taatccagag gttgattgtt ccagacgcgt tcaggcaccg
     7321 ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg
     7381 gtgaagccga gccgctcgta gaaggggagg ttgcggggcg cggaggtctc caggaaggcg
     7441 ggcaccccgg cgcgctcggc cgcctccact ccggggagca cgacggcgct gcccagaccc
     7501 ttgccctggt ggtcgggcga gacgccgacg gtggccagga accacgcggg ctccttgggc
     7561 cggtgcggcg ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga accgctcaac
     7621 tcggccatgc gcgggccgat ctcggcgaac accgcccccg cttcgacgct ctccggcgtg
     7681 gtccagaccg ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg
     7741 acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg gtccgggtcg
     7801 acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg tacggcccgg
     7861 gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat ggtcgcgttt
     7921 tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg
     7981 aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg
     8041 cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg
     8101 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     8161 acctggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     8221 gggactttcc acaccctaac tgacacacat tcgcgcgttt acccggggag catgtcaagg
     8281 tcaaaatcgt caagagcgtc agcaggcagc atatcaaggt caaagtcgtc aagggcatcg
     8341 gctgggagca tgtctaagtc aaaatcgtca agggcgtcgg tcggcccgcc gctttcgcac
     8401 tttagctgtt tctccaggcc acatatgatt agttccaggc cgaaaaggaa ggcaggttcg
     8461 gctccctgcc ggtcgaacag ctcaattgct tgtttcagaa gtgggggcat agaatcggtg
     8521 gtaggtgtct ctctttcctc ttttgctact tgatgctcct gttcctccaa tacgcagccc
     8581 agtgtaaagt ggcccacggc ggacagagcg tacagtgcgt tctccaggga gaagccttgc
     8641 tgacacagga acgcgagctg attttccagg gtttcgtact gtttctctgt tgggcgggtg
     8701 ccgagatgca ctttagcccc gtcgcgatgt gagaggagag cacagcggta tgacttggcg
     8761 ttgttccgca gaaagtcttg ccatgactcg ccttccaggg ggcaggagtg ggtatgatgc
     8821 ctgtccagca tctcgattgg cagggcatcg agcagggccc gcttgttctt cacgtgccag
     8881 tacagggtag gctgctcaac tcccagcttt tgagcgagtt tccttgtcgt caggccttcg
     8941 ataccgactc cattgagtaa ttccagagca gagtttatga ctttgctctt gtccagtcta
     9001 gacatggtaa ttcgatgatc cccctgggga gagaggtcgg tgattcggtc aacgagggag
     9061 ccgactgccg acgtgcgctc cggaggcttg cagaatgcgg aacaccgcgc gggcaggaac
     9121 agggcccaca ctaccgcccc acaccccgcc tcccgcaccg ccccttcccg gccgctgctc
     9181 tcggcgcgcc ctgctgagca gccgctattg gccacagccc atcgcggtcg gcgcgctgcc
     9241 attgctccct ggcgctgtcc gtctgcgagg gtactagtga gacgtgcggc ttccgtttgt
     9301 cacgtccggc acgccgcgaa ccgcaaggaa ccttcccgac ttaggggcgg agcaggaagc
     9361 gtcgccgggg ggcccacaag ggtagcggcg aagatccggg tgacgctgcg aacggacgtg
     9421 aagaatgtgc gagacccagg gtcggcgccg ctgcgtttcc cggaaccacg cccagagcag
     9481 ccgcgtccct gcgcaaaccc agggctgcct tggaaaaggc gcaaccccaa ccccgtggaa
     9541 ttatcacctc gagtttactc cctatcagtg atagagaacg tatgaagagt ttactcccta
     9601 tcagtgatag agaacgtatg cagactttac tccctatcag tgatagagaa cgtataagga
     9661 gtttactccc tatcagtgat agagaacgta tgaccagttt actccctatc agtgatagag
     9721 aacgtatcta cagtttactc cctatcagtg atagagaacg tatatccagt ttactcccta
     9781 tcagtgatag agaacgtata agctttgctt atgtaaacca gggcgcctat aaaagagtgc
     9841 tgattttttg agtaaacttc aattccacaa cacttttgtc ttataccaac tttccgtacc
     9901 acttcctacc ctcgtaaaga attcaccatg gcggcccccg gcagcgcccg gcgacccctg
     9961 ctgctgctac tgctgttgct gctgctcggc ctcatgcatt gtgcgtcagc agcaatgttt
    10021 atggtgaaaa atggcaacgg gaccgcgtgc ataatggcca acttctctgc tgccttctca
    10081 gtgaactacg acaccaagag tggccctaag aacatgacct ttgacctgcc atcagatgcc
    10141 acagtggtgc tcaaccgcag ctcctgtgga aaagagaaca cttctgaccc cagtctcgtg
    10201 attgcttttg gaagaggaca tacactcact ctcaatttca cgagaaatgc aacacgttac
    10261 agcgtccagc tcatgagttt tgtttataac ttgtcagaca cacacctttt ccccaatgcg
    10321 agctccaaag aaatcaagac tgtggaatct ataactgaca tcagggcaga tatagataaa
    10381 aaatacagat gtgttagtgg cacccaggtc cacatgaaca acgtgaccgt aacgctccat
    10441 gatgccacca tccaggcgta cctttccaac agcagcttca gcaggggaga gacacgctgt
    10501 gaacaagaca ggccttcccc aaccacagcg ccccctgcgc cacccagccc ctcgccctca
    10561 cccgtgccca agagcccctc tgtggacaag tacaacgtga gcggcaccaa cgggacctgc
    10621 ctgctggcca gcatggggct gcagctgaac ctcacctatg agaggaagga caacacgacg
    10681 gtgacaaggc ttctcaacat caaccccaac aagacctcgg ccagcgggag ctgcggcgcc
    10741 cacctggtga ctctggagct gcacagcgag ggcaccaccg tcctgctctt ccagttcggg
    10801 atgaatgcaa gttctagccg gtttttccta caaggaatcc agttgaatac aattcttcct
    10861 gacgccagag accctgcctt taaagctgcc aacggctccc tgcgagcgct gcaggccaca
    10921 gtcggcaatt cctacaagtg caacgcggag gagcacgtcc gtgtcacgaa ggcgttttca
    10981 gtcaatatat tcaaagtgtg ggtccaggct ttcaaggtgg aaggtggcca gtttggctct
    11041 gtggaggagt gtctgctgga cgagaacagc atgctgatcc ccatcgctgt gggtggtgcc
    11101 ctggcggggc tggtcctcat cgtcctcatc gcctacctcg tcggcaggaa gaggagtcac
    11161 gcaggctacc agactatcgg tggatctgga ggttctggtg gaatggtgag caagggcgag
    11221 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
    11281 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
    11341 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
    11401 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
    11461 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
    11521 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
    11581 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
    11641 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
    11701 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
    11761 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
    11821 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
    11881 gccgccggga tcactctcgg catggacgag ctgtacaagt ccggagcggc cgccaccatg
    11941 gactacaagg atgacgacga taagggaaag tcttacccaa ctgtga
//
