LOCUS       sequence-431745-       11730 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the human EndoTag (3xFLAG-EEA1) for EndoIPs. mTagBFP2 to 
            sort for even expression levels..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11730)
  AUTHORS   Hollingsworth LR, Veeraraghavan P, Paulo JA, Harper JW
  TITLE     Spatiotemporal proteomic profiling of cellular responses to NLRP3 
            agonists
  JOURNAL   bioRxiv 2024.04.19.590338
REFERENCE   2  (bases 1 to 11730)
  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 
            2024.04.19.590338"
COMMENT     Sequence Label: pLV-UBC mTagBFP2-P2A-3xFLAG-hEEA1
FEATURES             Location/Qualifiers
     source          1..11730
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(118..516)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     misc_feature    519..635
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(902..946)
                     /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    1131..1364
                     /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    1857..1982
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             2029..2209
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(2210..2436)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(2464..2484)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2464..2482)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2503..2519)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2503..2519)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2516..2538)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2527..2543
                     /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)."
     promoter        complement(join(2551..2557,2558..2575,2576..2581))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2596..2617
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2734..2751)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2905..3493)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2985..3004)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3664..4455,4456..4524))
                     /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     4287..4306
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4525..4629)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4655..5110)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4792..4813)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5004..5023
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5237..5259
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5251..5268
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5252..5268
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5278..5297
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5278..5296
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      5317..5452
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5492..5613
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5503..5522)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5557..5576
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             5685..5918
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    6005..6593
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6111..6122
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6520..6540
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6651..10886)
                     /codon_start=1
                     /product="human early endosome antigen 1"
                     /label=EEA1
                     /translation="MLRRILQRTPGRVGSQGSDLDSSATPINTVDVNNESSSEGFICPQ
                     CMKSLGSADELFKHYEAVHDAGNDSGHGGESNLALKRDDVTLLRQEVQDLQASLKEEKW
                     YSEELKKELEKYQGLQQQEAKPDGLVTDSSAELQSLEQQLEEAQTENFNIKQMKDLFEQ
                     KAAQLATEIADIKSKYDEERSLREAAEQKVTRLTEELNKEATVIQDLKTELLQRPGIED
                     VAVLKKELVQVQTLMDNMTLERERESEKLKDECKKLQSQYASSEATISQLRSELAKGPQ
                     EVAVYVQELQKLKSSVNELTQKNQTLTENLLKKEQDYTKLEEKHNEESVSKKNIQATLH
                     QKDLDCQQLQSRLSASETSLHRIHVELSEKGEATQKLKEELSEVETKYQHLKAEFKQLQ
                     QQREEKEQHGLQLQSEINQLHSKLLETERQLGEAHGRLKEQRQLSSEKLMDKEQQVADL
                     QLKLSRLEEQLKEKVTNSTELQHQLDKTKQQHQEQQALQQSTTAKLREAQNDLEQVLRQ
                     IGDKDQKIQNLEALLQKSKENISLLEKEREDLYAKIQAGEGETAVLNQLQEKNHTLQEQ
                     VTQLTEKLKNQSESHKQAQENLHDQVQEQKAHLRAAQDRVLSLETSVNELNSQLNESKE
                     KVSQLDIQIKAKTELLLSAEAAKTAQRADLQNHLDTAQNALQDKQQELNKITTQLDQVT
                     AKLQDKQEHCSQLESHLKEYKEKYLSLEQKTEELEGQIKKLEADSLEVKASKEQALQDL
                     QQQRQLNTDLELRATELSKQLEMEKEIVSSTRLDLQKKSEALESIKQKLTKQEEEKKIL
                     KQDFETLSQETKIQHEELNNRIQTTVTELQKVKMEKEALMTELSTVKDKLSKVSDSLKN
                     SKSEFEKENQKGKAAILDLEKTCKELKHQLQVQMENTLKEQKELKKSLEKEKEASHQLK
                     LELNSMQEQLIQAQNTLKQNEKEEQQLQGNINELKQSSEQKKKQIEALQGELKIAVLQK
                     TELENKLQQQLTQAAQELAAEKEKISVLQNNYEKSQETFKQLQSDFYGRESELLATRQD
                     LKSVEEKLSLAQEDLISNRNQIGNQNKLIQELKTAKATLEQDSAKKEQQLQERCKALQD
                     IQKEKSLKEKELVNEKSKLAEIEEIKCRQEKEITKLNEELKSHKLESIKEITNLKDAKQ
                     LLIQQKLELQGKADSLKAAVEQEKRNQQILKDQVKKEEEELKKEFIEKEAKLHSEIKEK
                     EVGMKKHEENEAKLTMQITALNENLGTVKKEWQSSQRRVSELEKQTDDLRGEIAVLEAT
                     VQNNQDERRALLERCLKGEGEIEKLQTKVLELQRKLDNTTAAVQELGRENQSLQIKHTQ
                     ALNRKWAEDNEVQNCMACGKGFSVTVRRHHCRQCGNIFCAECSAKNALTPSSKKPVRVC
                     DACFNDLQG"
     protein_bind    complement(10892..10916)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(10932..10955)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(10959..10982)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(10986..11009)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(11013..11069)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(join(11079..11730,1..47))
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
ORIGIN
        1 tccatgtaca gcttcatgtg catgttctcc ttaatcagct cgctcatggt ggcgaccggt
       61 ggatccgcta gcgtcggtgt cttctatgga ggtcaaaaca gcgtggatgg cgtctcccca
      121 agtgacgatc acagcgatcc acaaacaaga accgcgaccc aaatcccggc tgcgacggaa
      181 ctagctgtgc cacacccggc gcgtccttat ataatcatcg gcgttcaccg ccccacggag
      241 atccctccgc agaatcgccg agaagggact acttttcctc gcctgttccg ctctctggaa
      301 agaaaaccag tgccctagag tcacccaagt cccgtcctaa aatgtccttc tgctgatact
      361 ggggttctaa ggccgagtct tatgagcagc gggccgctgt cctgagcgtc cgggcggaag
      421 gatcaggacg ctcgctgcgc ccttcgtctg acgtggcagc gctcgccgtg aggagggggg
      481 cgcccgcggg aggcgccaaa acccggcgcg gaggccataa aattttgaat tttgtaattt
      541 gtttttgtaa ttctttagtt tgtatgtctg ttgctattat gtctactatt ctttcccctg
      601 cactataccc cccaatcccc ccttttcttt taaaagttaa ccgataccgt cgagatccgt
      661 tcactaatcg aatggatctg tctctgtctc tctctccacc ttcttcttct attccttcgg
      721 gcctgtcggg tcccctcggg gttgggaggt gggtctgaaa cgataatggt gaatatccct
      781 gcctaactct attcactata gaaagtacag caaaaactat tcttaaacct accaagcctc
      841 ctactatcat tatgaataat tttatatacc acagccaatt tgttatgtta aaccaattcc
      901 acaaacttgc ccatttatct aattccaata attcttgttc attcttttct tgctggtttt
      961 gcgattcttc aattaaggag tgtattaagc ttgtgtaatt gttaatttct ctgtcccact
     1021 ccatccaggt cgtgtgattc caaatctgtt ccagagattt attactccaa ctagcattcc
     1081 aaggcacagc agtggtgcaa atgagttttc cagagcaacc ccaaatcccc aggagctgtt
     1141 gatcctttag gtatctttcc acagccagga ttcttgcctg gagctgcttg atgccccaga
     1201 ctgtgagttg caacagatgc tgttgcgcct caatagccct cagcaaattg ttctgctgct
     1261 gcactatacc agacaataat tgtctggcct gtaccgtcag cgtcattgac gctgcgccca
     1321 tagtgcttcc tgctgctccc aagaacccaa ggaacaaagc tcctattccc actgctcttt
     1381 tttctctctg caccactctt ctctttgcct tggtgggtgc tactcctaat ggttcaattt
     1441 ttactacttt atatttatat aattcacttc tccaattgtc cctcatatct cctcctccag
     1501 gtctgaagat cagcggccgc ttgctgtgcg gtggtcttac ttttgttttg ctcttcctct
     1561 atcttgtcta aagcttcctt ggtgtctttt atctctatcc tttgatgcac acaatagagg
     1621 gttgctactg tattatataa tgatctaagt tcttctgatc ctgtctgaag ggatggttgt
     1681 agctgtccca gtatttgtct acagccttct gatgtttcta acaggccagg attaactgcg
     1741 aatcgttcta gctccctgct tgcccatact atatgtttta atttatattt tttctttccc
     1801 cctggcctta accgaatttt ttcccatcgc gatctaattc tcccccgctt aatactgacg
     1861 ctctcgcacc catctctctc cttctagcct ccgctagtca aaatttttgg cgtactcacc
     1921 agtcgccgcc cctcgcctct tgccgtgcgc gcttcagcaa gccgagtcct gcgtcgagag
     1981 agctcctctg gtttcccttt cgctttcaag tccctgttcg ggcgccactg ctagagattt
     2041 tccacactga ctaaaagggt ctgagggatc tctagttacc agagtcacac aacagacggg
     2101 cacacactac ttgaagcact caaggcaagc tttattgagg cttaagcagt gggttcccta
     2161 gttagccaga gagctcccag gctcagatct ggtctaacca gagagacccg tttatgtatc
     2221 gagctaggca cttaaataca atatctctgc aatgcggcaa ttcagtggtt cgtccaatcc
     2281 atgtcagacc cgtctgttgc cttcctaata aggcacgatc gtaccacctt acttccacca
     2341 atcggcatgc acggtgcttt ttctctcctt gtaaggcatg ttgctaactc atcgttacca
     2401 tgttgcaaga ctacaagagt attgcataag actacattaa gcttgcagct ccagcttttg
     2461 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt
     2521 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtggaa
     2581 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     2641 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     2701 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     2761 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2821 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2881 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2941 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3001 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3061 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3121 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3181 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3241 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3301 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3361 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3421 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3481 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3541 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3601 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3661 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3721 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     3781 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     3841 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     3901 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     3961 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4021 attcagctcc ggttcccaac gatcaaggcg agttacatga tctcccatgt tgtgcaaaaa
     4081 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4141 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4201 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4261 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4321 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4381 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4441 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4501 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4561 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4621 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     4681 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4741 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4801 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4861 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4921 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4981 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     5041 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     5101 cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     5161 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     5221 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa
     5281 tacgactcac tatagggcga attgggtacg tccctaggcc tccaaaaaag cctcctcact
     5341 acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta
     5401 gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag
     5461 ttaggggcgg gatagctaga gccagacatg ataagataca ttgatgagtt tggacaaacc
     5521 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     5581 tttgtaacca ttataagctg caataaacaa gttcctctca ctctctgata ttcatttctt
     5641 tgcaagttat aaatactgaa taataagatg acatgaacta ctactgctag agattttcca
     5701 cactgactaa aagggtctga gggatctcta gttaccagag tcacacaaca gacgggcaca
     5761 cactacttga agcactcaag gcaagcttta ttgaggctta agcagtgggt tccctagtta
     5821 gccagagagc tcccaggctc agatctggtc taaccagaga gacccagtac aagcaaaaag
     5881 cagatcttgt cttcgttggg agtgaattag cccttccagt cccccctttt cttttaaaaa
     5941 gtggctaaga tctacagctg ccttgtaagt cattggtctt aaaggtaccg agctcgaatt
     6001 ccaggcgggg aggcggccca aagggagatc cgactcgtct gagggcgaag gcgaagacgc
     6061 ggaagaggcc gcagagccgg cagcaggccg cgggaaggaa ggtccgctgg attgagggcc
     6121 gaagggacgt agcagaagga cgtcccgcgc agaatccagg tggcaacaca ggcgagcagc
     6181 catggaaagg acgtcagctt ccccgacaac accacggaat tgtcagtgcc caacagccga
     6241 gcccctgtcc agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc aatagggagg
     6301 gggaaagcga aagtcccgga aaggagctga caggtggtgg caatgcccca accagtgggg
     6361 gttgcgtcag caaacacagt gcacaccacg ccacgttgcc tgacaacggg ccacaactcc
     6421 tcataaagag acagcaacca ggatttatac aaggaggaga aaatgaaagc catacgggaa
     6481 gcaatagcat gatacaaagg cattaaagca gcgtatccac atagcgtaaa aggagcaaca
     6541 tagttaagaa taccagtcaa tctttcacaa attttgtaat ccagaggttg attgtcgacg
     6601 cggccgcttt atccttgcaa gtcattgaaa cgattgtcga cgcggccgct ttatccttgc
     6661 aagtcattga aacatgcatc acagacacga acaggcttct tggaggaagg agttaaggca
     6721 tttttggctg aacattcagc acagaagata tttccacact gtcggcagtg atgccgtctc
     6781 actgttactg aaaagccttt cccacaggcc atacagtttt gtacttcatt gtcttcggcc
     6841 cactttctat tcaacgcttg tgtatgtttg atctgaagtg attggttttc tctgcccagc
     6901 tcctgcactg ctgcagttgt attatccagc tttctttgca attctaatac tttggtttga
     6961 agcttttcta tttcaccttc tcctttaaga catctttcca gtagtgctct cctttcatct
     7021 tgattattct gaaccgttgc ttctaatact gcaatttcac cccgtaagtc atccgtttgt
     7081 ttctcaagct cactaactct ccgttgacta gattgccact ccttcttcac agtgcctaag
     7141 ttttcattta atgctgtaat ctgcatggta agtttagcct cattttcttc atgcttcttc
     7201 attcctactt ccttttcttt tatttcggaa tgcaacttag cttctttctc aataaattct
     7261 ttcttcagct cctcttcttc ctttttcacc tggtctttta gtatctgctg atttctcttc
     7321 tcctgttcaa cagctgcctt cagggagtcc gcttttcctt gaagttctaa tttctgctga
     7381 attagaagct gtttagcatc tttaagattt gttatctcct ttatgctttc tagtttgtgg
     7441 gacttgagtt cttcgtttag tttagtgatt tctttttctt gtctacattt aatttcttct
     7501 atctctgcca atttagactt ctcatttacc agttcttttt ctttcagtga cttttctttc
     7561 tgaatgtctt gtagtgcttt acatcgctcc tgcaattgct gttctttctt tgctgaatcc
     7621 tgctccaatg tagccttggc agtcttcagt tcttgaatca atttattttg atttccaatt
     7681 tgatttctgt ttgaaatcaa gtcctcctgt gctagagaaa gcttctcttc tacagactta
     7741 agatcttgcc tggtggctag aagttcagat tccctcccat agaaatcaga ttgaagctgt
     7801 ttgaaagttt cctgactttt ttcatagttg ttttgtaata ctgatatttt ctctttctct
     7861 gctgcaagtt cctgggctgc ctgtgttaac tgctgctgta gtttattctc aagctctgtc
     7921 ttctgtaaaa cagcaatttt aagctctcct tggagtgctt caatttgttt tttcttctgt
     7981 tcacttgatt gctttagctc atttatgttc ccctgaagtt gttgctcttc tttttcattt
     8041 tgttttaaag tattctgggc ctgtataagt tgttcctgca ttgaattgag ttccaatttc
     8101 aactgatgag aagcctcctt ctctttttca agtgactttt tcagttcctt ctgttcctta
     8161 agtgtgtttt ccatctgcac ttgaagttga tgctttaatt ctttgcaagt tttttccaag
     8221 tctaatatag cggcttttcc tttctgattc tccttttcaa attcactttt agagtttttc
     8281 aaagaatcag aaacttttga tagtttgtcc tttactgtag aaagctctgt cattaaagct
     8341 tctttctcca ttttcacttt ttgtagttct gttactgttg tttgaattct gttattcaat
     8401 tcctcatgct gaatctttgt ttcttgactt aaagtttcaa aatcttgttt caggattttt
     8461 ttttcttcct cttgcttggt aagcttttgc ttgatacttt caagggcttc agattttttc
     8521 tgtagatcca atcttgtact ggatactatt tccttctcca tttcaagttg tttactcaat
     8581 tctgtggctc tgagctctaa atctgtgttc agctgtcttt gctgttgtag atcttgcaaa
     8641 gcctgctcct tgcttgcttt aacttcaaga ctatcagctt ctagtttctt aatttgacct
     8701 tctagctctt cggttttctg ttctaaagag aggtatttct ctttatattc tttaagatga
     8761 ctttccagct gactgcaatg ttcttgcttg tcttgtaact ttgcagtgac ctgatccaac
     8821 tgagtagtaa tcttatttaa ctcctgctgt ttatcttgta atgcattttg agctgtgtcc
     8881 aaatgattct gaagatcagc tctttgagca gtttttgctg cttctgctga tagtaatagt
     8941 tcggttttgg ctttaatctg tatgtcaagc tgggagacct tctccttgct ttcatttaat
     9001 tgactattta attcattgac actagtttct agggaaagga cacggtcttg tgcagctcta
     9061 agatgtgcct tctgctcttg tacctggtca tgcaaattct cctgggcttg tttatgactt
     9121 tctgactgat tcttcagctt ctctgttagt tgagttactt gctcctgtag tgtatggttt
     9181 ttttcttgta actggttaag aacagcagtc tctccttcac cagcctgaat ttttgcataa
     9241 agatcttctc tttctttttc tagtaatgaa atattttctt tactcttctg taataaagct
     9301 tcaaggttct ggatcttttg gtccttatcg ccaatttgac gtagaacttg ttccaaatca
     9361 ttctgagctt ctcgaagttt tgccgtggtg ctttgctgaa gagcctgttg ttcttgatgc
     9421 tgttgctttg ttttatctaa ttgatgctgc aattctgtag aatttgtaac tttttccttc
     9481 aactgctctt ctaaccgaga aagtttgagt tgtaaatcag ccacttgttg ttctttatcc
     9541 atcaactttt cacttgaaag ctgtctctgt tccttcagcc taccatgagc ttcccctagt
     9601 tggcgctctg tctccagaag tttgctatgt aattgattaa tttcactttg gagttgtaac
     9661 ccatgctgct ccttttcttc tctctgttgt tgtagctgct taaactccgc ctttagatgc
     9721 tggtacttgg tctctacctc agataattct tctttgagct tttgagtagc ttctcctttt
     9781 tcacttagtt ctacatgtat tctatgcagt gaggtttcag atgcagacaa tcttgactga
     9841 agctgttgac aatctaggtc tttttgatga agggttgcct gaatattctt tttactcaca
     9901 gattcttcat tatgtttctc ctctaactta gtatagtctt gttctttttt cagcaagttt
     9961 tctgtcaagg tctgattttt ttgtgttaat tcattaactg aacttttcag tttttgtagt
    10021 tcctgtacat atacagcaac ttcctggggg cctttggcaa gttcactcct tagctggctt
    10081 attgtggcct ctgagctagc atattgtgac tgcaattttt tgcattcatc tttgagtttt
    10141 tcagattctc gctcacgttc caaggtcatg ttatccatta gtgtttgaac ttggaccagt
    10201 tctttcttta gcacggcaac atcttctata ccaggtctct gaagcagttc cgtcttcaga
    10261 tcttgaatta cagttgcctc tttgtttaat tcttctgtca gacgtgtcac tttttgttca
    10321 gcagcttctc gaagactcct ttcttcatca tactttgact ttatatctgc aatttcagta
    10381 gcaagttggg ctgctttctg ttcaaataag tctttcattt gcttaatatt aaaattttct
    10441 gtttgggctt cttctaattg ctgttccaaa gactgtagtt ctgctgatga atcagtcacc
    10501 aacccatcag gtttggcctc ttgctgctgc agcccttgat atttttctaa ttccttcttt
    10561 aattcttccg agtaccattt ttcttcctta agtgaagcct gtaggtcttg gacctcttgt
    10621 ctgagcagtg ttacatcatc tcgcttcaaa gcaagattag actctcctcc atgacctgag
    10681 tcattaccag catcatgaac agcttcataa tgtttgaaaa gttcatcagc agatccaaga
    10741 gatttcatac actggggaca tatgaaaccc tctgaagagc tttcattatt gacgtccact
    10801 gtgtttatag gagttgctga tgaatctaaa tcagaacctt gagagccaac tctcccagga
    10861 gtcctctgta aaatcctcct taacatggtg gagcctgctt ttttgtacaa acttgtttga
    10921 taacctcgag ccttatcgtc gtcatccttg taatctccct tatcgtcgtc atccttgtaa
    10981 tcgcccttgt catcgtcatc cttgtagtcc atcggtccag gattctcttc gacatctccg
    11041 gcttgtttca gcagagagaa gtttgttgcg ccggatccat taagcttgtg ccccagtttg
    11101 ctagggaggt cgcagtatct ggccactgcc acctcgtgct gctcgacgta ggtctcgttg
    11161 ttggcctcct tgattctttc cagtctgtag tccacatagt agacgccagg catcttgagg
    11221 ttcttagcgg gtttcttgga tctatatgtg gtcttggcgt ttgcgatcag atggctcccg
    11281 cccacgagct tcagggccat gtcgtttctg ccttccaggc cgccgtcagc ggggtacagc
    11341 gtctcggtga aggcctccca gccgagtgtt ttcttctgca tcacagggcc gttggatgtg
    11401 aagttcaccc ctctgatctt gacgttgtag atgaggcagc cgtcctggag gctggtgtcc
    11461 tgggtagcgg tcagcacgcc cccgtcttcg tatgtggtga ctctctccca tgtgaagccc
    11521 tcagggaagg actgcttgaa gaagtcgggg atgccctggg tgtggttgat gaaggtcttg
    11581 ctgccgtaga ggaagctagt agccaggatg tcgaaggcga aggggagagg gccgccctcg
    11641 accaccttga ttctcatggt ctgggtgccc tcgtagggct tgccttcgcc ctcggatgtg
    11701 cacttgaagt gatggttgtc cacggtgccc
//
