LOCUS       sequence-324201-       13469 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  All-in-One piggyBac transposon Gateway Destination vector for 
            dox-inducible expression of N-terminal Halo tagged protein 
            (inducible GFP and constitutive rtTA and neomycin resistance).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13469)
  AUTHORS   Banerjee AK, Blanco MR, Bruce EA, Honson DD, Chen LM, Chow A, Bhat 
            P, Ollikainen N, Quinodoz SA, Loney C, Thai J, Miller ZD, Lin AE, 
            Schmidt MM, Stewart DG, Goldfarb D, De Lorenzo G, Rihn SJ, Voorhees 
            RM, Botten JW, Majumdar D, Guttman M
  TITLE     SARS-CoV-2 Disrupts Splicing, Translation, and Protein Trafficking 
            to Suppress Host Defenses.
  JOURNAL   Cell. 2020 Nov 25;183(5):1325-1339.e21. doi: 
            10.1016/j.cell.2020.10.004. Epub 2020 Oct 8.
  PUBMED    33080218
REFERENCE   2  (bases 1 to 13469)
  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"; doi: 
            "10.1016/j.cell.2020.10.004"; journalName: "Cell"; date: 
            "2020-11-25- 25"; volume: "183"; issue: "5"; pages: "1325-1339.e21"
COMMENT     Sequence Label: pB-Halo,IRES-eGFP
FEATURES             Location/Qualifiers
     source          1..13469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(28..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     583..605
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     597..614
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     598..614
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        621..639
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   706..768
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    1140..1158
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1177..1195
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1214..1232
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1251..1269
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1288..1306
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1325..1343
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1362..1380
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1399..1417
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1439..1463
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1581..2471
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             2484..2504
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     protein_bind    2529..2653
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2678..2683,2684..2701,2702..2708)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2762..3421
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2828..2847)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3784..4089
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3968..3987
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4130..4254)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    join(4296..4870,4871..4873,4874..4882)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4463..4480)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4690..4709
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4883..4885,4886..4888,4889..5602)
                     /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(4883..4885,4886..4888,4889..5599)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4928..4949)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5189..5208)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5536..5557
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5614..5633)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5679..5734
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5733..5752)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     enhancer        6177..6556
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6671..7976
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    7013..7944
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     CDS             join(7993..8610,8611..8616,8617..8652,8653..8655,8656..8691,
                     8692..8694,8695..8730,8731..8739)
                     /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
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    8766..9210
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        9243..9289
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        9281..9299
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             9309..10103
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             9309..10103
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(9363..9382)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9973..9992
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    10150..10722
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     repeat_region   11096..11130
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(11279..11297)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11318..11334)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11318..11334)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11331..11353)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11342..11358
                     /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(11366..11372,11373..11390,11391..11396))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11411..11432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11549..11566)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11720..12308)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11800..11819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12479..13270,13271..13339))
                     /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     13102..13121
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13340..13444)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
      541 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
      601 aaacgacggc cagtgaattg taatacgact cactataggg cgaattggag ctcggtattc
      661 acgacagcag gctgaataat aaaaaaatta gaaactatta tttaacccta gaaagataat
      721 catattgtga cgtacgttaa agataatcat gcgtaaaatt gacgcatgtg ttttatcggt
      781 ctgtatatcg aggtttattt attaatttga atagatatta agttttatta tatttacact
      841 tacatactaa taataaattc aacaaacaat ttatttatgt ttatttattt attaaaaaaa
      901 aacaaaaact caaaatttct tctataaagt aacaaaactt ttaaacattc tctcttttac
      961 aaaaataaac ttattttgta ctttaaaaac agtcatgttg tattataaaa taagtaatta
     1021 gcttaaccta tacataatag aaacaaatta tacttattag tcagtcagaa acaactttgg
     1081 cacatatcaa tattatgctc tcgttaatta actcgaggga tccaagctgg ccgcacgtct
     1141 ccctatcagt gatagagaag tcgacacgtc tcgagctccc tatcagtgat agagaaggta
     1201 cgtctagaac gtctccctat cagtgataga gaagtcgaca cgtctcgagc tccctatcag
     1261 tgatagagaa ggtacgtcta gaacgtctcc ctatcagtga tagagaagtc gacacgtctc
     1321 gagctcccta tcagtgatag agaaggtacg tctagaacgt ctccctatca gtgatagaga
     1381 agtcgacacg tctcgagctc cctatcagtg atagagaagg taccccctat ataagcagag
     1441 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     1501 gaagacaccg ggaccgatcc agcctccgcg gtggcggcca tcacaagttt gactagtacc
     1561 ggttactcga ggctagcacc atggcagaaa tcggtactgg ctttccattc gacccccatt
     1621 atgtggaagt cctgggcgag cgcatgcact acgtcgatgt tggtccgcgc gatggcaccc
     1681 ctgtgctgtt cctgcacggt aacccgacct cctcctacgt gtggcgcaac atcatcccgc
     1741 atgttgcacc gacccatcgc tgcattgctc cagacctgat cggtatgggc aaatccgaca
     1801 aaccagacct gggttatttc ttcgacgacc acgtccgctt catggatgcc ttcatcgaag
     1861 ccctgggtct ggaagaggtc gtcctggtca ttcacgactg gggctccgct ctgggtttcc
     1921 actgggccaa gcgcaatcca gagcgcgtca aaggtattgc atttatggag ttcatccgcc
     1981 ctatcccgac ctgggacgaa tggccagaat ttgcccgcga gaccttccag gccttccgca
     2041 ccaccgacgt cggccgcaag ctgatcatcg atcagaacgt ttttatcgag ggtacgctgc
     2101 cgatgggtgt cgtccgcccg ctgactgaag tcgagatgga ccattaccgc gagccgttcc
     2161 tgaatcctgt tgaccgcgag ccactgtggc gcttcccaaa cgagctgcca atcgccggtg
     2221 agccagcgaa catcgtcgcg ctggtcgaag aatacatgga ctggctgcac cagtcccctg
     2281 tcccgaagct gctgttctgg ggcaccccag gcgttctgat cccaccggcc gaagccgctc
     2341 gcctggccaa aagcctgcct aactgcaagg ctgtggacat cggcccgggt ctgaatctgc
     2401 tgcaagaaga caacccggac ctgatcggca gcgagatcgc gcgctggctg tcgacgctcg
     2461 agatttccgg cgagccaacc actgaggatc tgtactttca gagcgataac gcgatcgccg
     2521 gcgcgccaac aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat ataaatatca
     2581 atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa acacaacata
     2641 tccagtcact atggcggccg cattaggcac cccaggcttt acactttatg cttccggctc
     2701 gtataatgtg tggattttga gttaggatcc gtcgagattt tcaggagcta aggaagctaa
     2761 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
     2821 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
     2881 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
     2941 tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa tgaaagacgg
     3001 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
     3061 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
     3121 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
     3181 gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     3241 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
     3301 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
     3361 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     3421 aagatctgga tcccgacaat caacgcgtgg atccggctta ctaaaagcca gataacagta
     3481 tgcgtatttg cgcgctgatt tttgcggtat aagaatatat actgatatgt atacccgaag
     3541 tatgtcaaaa agaggtatgc tatgaagcag cgtattacag tgacagttga cagcgacagc
     3601 tatcagttgc tcaaggcata tatgatgtca atatctccgg tctggtaagc acaaccatgc
     3661 agaatgaagc ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg
     3721 ctgaggtcgc ccggtttatt gaaatgaacg gctcttttgc tgacgagaac aggggctggt
     3781 gaaatgcagt ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat
     3841 gtacagagtg atattattga cacgcccggg cgacggatgg tgatccccct ggccagtgca
     3901 cgtctgctgt cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa
     3961 agctggcgca tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa
     4021 gtggctgatc tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg
     4081 ggaatataaa tgtcaggctc ccttatacac agccagtctg caggtcgacc atagtgactg
     4141 gatatgttgt gttttacagt attatgtagt ctgtttttta tgcaaaatct aatttaatat
     4201 attgatattt atatcatttt acgtttctcg ttcagctttc ttgtacaaag tggtgatcta
     4261 gctaaaacgc ggcctcgaat cgaattccgc ccccccccct ctccctcccc cccccctaac
     4321 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     4381 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     4441 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     4501 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     4561 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     4621 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     4681 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     4741 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     4801 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     4861 cgatgataat atggccacaa ccatggtgag caagggcgag gagctgttca ccggggtggt
     4921 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     4981 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     5041 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     5101 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     5161 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     5221 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     5281 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     5341 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     5401 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     5461 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     5521 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     5581 catggacgag ctgtacaagt aaagcggccg cgatcttttt ccctctgcca aaaattatgg
     5641 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     5701 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     5761 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     5821 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     5881 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     5941 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     6001 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc
     6061 cctcgacctg caggaattcg attacttaag cctaggatcc agatctgtac attcgaatcg
     6121 atgaattcgt taaccatgga agcttctgca gcggccgcac tagcggtggc ggcctcgaca
     6181 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     6241 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     6301 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     6361 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     6421 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     6481 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     6541 catcgctatt accatgatga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     6601 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttgact
     6661 agtggagccg agagtaattc atacaaaagg agggatcgcc ttcgcaaggg gagagcccag
     6721 ggaccgtccc taaattctca cagacccaaa tccctgtagc cgccccacga cagcgcgagg
     6781 agcatgcgcc cagggctgag cgcgggtaga tcagagcaca caagctcaca gtccccggcg
     6841 gtggggggag gggcgcgctg agcgggggcc agggagctgg cgcggggcaa actgggaaag
     6901 tggtgtcgtg tgctggctcc gccctcttcc cgagggtggg ggagaacggt atataagtgc
     6961 ggtagtcgcc ttggacgttc tttttcgcaa cgggtttgcc gtcagaacgc aggtgagtgg
     7021 cgggtgtggc ttccgcgggc cccggagctg gagccctgct ctgagcgggc cgggctgata
     7081 tgcgagtgtc gtccgcaggg tttagctgtg agcattccca cttcgagtgg cgggcggtgc
     7141 gggggtgaga gtgcgaggcc tagcggcaac cccgtagcct cgcctcgtgt ccggcttgag
     7201 gcctagcgtg gtgtccgccg ccgcgtgcca ctccggccgc actatgcgtt ttttgtcctt
     7261 gctgccctcg attgccttcc agcagcatgg gctaacaaag ggagggtgtg gggctcactc
     7321 ttaaggagcc catgaagctt acgttggata ggaatggaag ggcaggaggg gcgactgggg
     7381 cccgcccgcc ttcggagcac atgtccgacg ccacctggat ggggcgaggc ctgtggcttt
     7441 ccgaagcaat cgggcgtgag tttagcctac ctgggccatg tggccctagc actgggcacg
     7501 gtctggcctg gcggtgccgc gttcccttgc ctcccaacaa gggtgaggcc gtcccgcccg
     7561 gcaccagttg cttgcgcgga aagatggccg ctcccggggc cctgttgcaa ggagctcaaa
     7621 atggaggacg cggcagcccg gtggagcggg cgggtgagtc acccacacaa aggaagaggg
     7681 ccttgcccct cgccggccgc tgcttcctgt gaccccgtgg tctatcggcc gcatagtcac
     7741 ctcgggcttc tcttgagcac cgctcgtcgc ggcgggggga ggggatctaa tggcgttgga
     7801 gtttgttcac atttggtggg tggagactag tcaggccagc ctggcgctgg aagtcattct
     7861 tggaatttgc ccctttgagt ttggagcgag gctaattctc aagcctctta gcggttcaaa
     7921 ggtattttct aaacccgttt ccaggtgttg tgaaagccac cgctaattca aagcaatccg
     7981 gagtaattca ccatgtctag actggacaag agcaaagtca taaacggcgc tctggaatta
     8041 ctcaatggag tcggtatcga aggcctgacg acaaggaaac tcgctcaaaa gctgggagtt
     8101 gagcagccta ccctgtactg gcacgtgaag aacaagcggg ccctgctcga tgccctgcca
     8161 atcgagatgc tggacaggca tcatacccac ttctgccccc tggaaggcga gtcatggcaa
     8221 gactttctgc ggaacaacgc caagtcattc cgctgtgctc tcctctcaca tcgcgacggg
     8281 gctaaagtgc atctcggcac ccgcccaaca gagaaacagt acgaaaccct ggaaaatcag
     8341 ctcgcgttcc tgtgtcagca aggcttctcc ctggagaacg cactgtacgc tctgtccgcc
     8401 gtgggccact ttacactggg ctgcgtattg gaggaacagg agcatcaagt agcaaaagag
     8461 gaaagagaga cacctaccac cgattctatg cccccacttc tgagacaagc aattgagctg
     8521 ttcgaccggc agggagccga acctgccttc cttttcggcc tggaactaat catatgtggc
     8581 ctggagaaac agctaaagtg cgaaagcggc gggccggccg acgcccttga cgattttgac
     8641 ttagacatgc tcccagccga tgcccttgac gactttgacc ttgatatgct gcctgctgac
     8701 gctcttgacg attttgacct tgacatgctc cccgggtaac taagtaagga tcccgtacgc
     8761 ctaggagcag gtttccccaa tgacacaaaa cgtgcaactt gaaactccgc ctggtctttc
     8821 caggtctaga ggggtaacac tttgtactgc gtttggctcc acgctcgatc cactggcgag
     8881 tgttagtaac agcactgttg cttcgtagcg gagcatgacg gccgtgggaa ctcctccttg
     8941 gtaacaagga cccacggggc caaaagccac gcccacacgg gcccgtcatg tgtgcaaccc
     9001 cagcacggcg actttactgc gaaacccact ttaaagtgac attgaaactg gtacccacac
     9061 actggtgaca ggctaaggat gcccttcagg taccccgagg taacacgcga cactcgggat
     9121 ctgagaaggg gactggggct tctataaaag cgctcggttt aaaaagcttc tatgcctgaa
     9181 taggtgaccg gaggtcggca cctttccttt gcaattactg accctatgaa tacactgact
     9241 gtttgacaat taatcatcgg catagtatat cggcatagta taatacgact cactatagga
     9301 gggccaccat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     9361 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     9421 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     9481 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     9541 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     9601 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     9661 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     9721 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     9781 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     9841 tgcccgacgg cgaggatctc gtcgtgacac atggcgatgc ctgcttgccg aatatcatgg
     9901 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     9961 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
    10021 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
    10081 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgaa
    10141 ttcgctagga ttatccctaa tacctgccac cccactctta atcagtggtg gaagaacggt
    10201 ctcagaactg tttgtttcaa ttggccattt aagtttagta gtaaaagact ggttaatgat
    10261 aacaatgcat cgtaaaacct tcagaaggaa aggagaatgt tttgtggacc actttggttt
    10321 tcttttttgc gtgtggcagt tttaagttat tagtttttaa aatcagtact ttttaatgga
    10381 aacaacttga ccaaaaattt gtcacagaat tttgagaccc attaaaaaag ttaaatgaga
    10441 aacctgtgtg ttcctttggt caacaccgag acatttaggt gaaagacatc taattctggt
    10501 tttacgaatc tggaaacttc ttgaaaatgt aattcttgag ttaacacttc tgggtggaga
    10561 atagggttgt tttcccccca cataattgga aggggaagga atatcattta aagctatggg
    10621 agggttgctt tgattacaac actggagaga aatgcagcat gttgctgatt gcctgtcact
    10681 aaaacaggcc aaaaactgag tccttgggtt gcatagaaag ctgccaaacg ctagccaaca
    10741 agctcgtcat cgctttgcag aagagcagag aggatatgct catcgtctaa agaactaccc
    10801 attttattat atattagtca cgatatctat aacaagaaaa tatatatata ataagttatc
    10861 acgtaagtag aacatgaaat aacaatataa ttatcgtatg agttaaatct taaaagtcac
    10921 gtaaaagata atcatgcgtc attttgactc acgcggtcgt tatagttcaa aatcagtgac
    10981 acttaccgca ttgacaagca cgcctcacgg gagctccaag cggcgactga gatgtcctaa
    11041 atgcacagcg acggattcgc gctatttaga aagagagagc aatatttcaa gaatgcatgc
    11101 gtcaatttta cgcagactat ctttctaggg ttaaaaaaga tttgcgcttt actcgaccta
    11161 aactttaaac acgtcataga atcttcgttt gacaaaaacc acattgtggc caagctgtgt
    11221 gacgcgacgc gcgctaaaga atggcaaacc aagtcgcgcg aggtaccagc ttttgttccc
    11281 tttagtgagg gttaatttcg agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
    11341 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
    11401 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
    11461 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
    11521 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
    11581 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
    11641 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
    11701 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
    11761 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
    11821 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
    11881 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
    11941 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
    12001 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
    12061 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
    12121 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
    12181 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
    12241 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
    12301 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
    12361 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
    12421 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
    12481 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
    12541 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
    12601 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
    12661 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
    12721 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
    12781 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
    12841 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
    12901 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
    12961 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
    13021 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
    13081 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
    13141 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
    13201 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
    13261 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
    13321 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
    13381 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
    13441 cgcgcacatt tccccgaaaa gtgccacct
//
