LOCUS       sequence-319399-       13631 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 NGFR and constitutive rtTA and neomycin resistance).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13631)
  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 13631)
  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-NGFR
FEATURES             Location/Qualifiers
     source          1..13631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(46..634)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(126..145)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(805..1596,1597..1665))
                     /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     1428..1447
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1666..1770)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1799..2254)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1936..1957)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2148..2167
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2384..2406
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2398..2415
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2399..2415
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2422..2440
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   2507..2569
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    2941..2959
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2978..2996
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3015..3033
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3052..3070
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3089..3107
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3126..3144
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3163..3181
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3200..3218
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3240..3264
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3382..4272
                     /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             4285..4305
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     protein_bind    4330..4454
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(4479..4484,4485..4502,4503..4509)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             4563..5222
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4629..4648)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             5585..5890
                     /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     5769..5788
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(5931..6055)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    join(6097..6671,6672..6674,6675..6683)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6264..6281)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6491..6510
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(7571..7590)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7636..7691
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7690..7709)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     enhancer        8132..8511
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8626..9931
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    8968..9899
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     CDS             join(9949..10566,10567..10572,10573..10608,10609..10611,
                     10612..10647,10648..10650,10651..10686,10687..10695)
                     /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    10722..11166
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        11199..11245
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        11237..11255
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             11265..12059
                     /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             11265..12059
                     /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(11319..11338)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     11929..11948
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    12106..12678
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     repeat_region   13052..13086
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(13236..13254)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(13275..13291)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13275..13291)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13288..13310)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13299..13315
                     /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(13323..13329,13330..13347,13348..13353))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13368..13389
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13506..13523)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
       61 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      121 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      181 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      241 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      301 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      361 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
      421 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
      481 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
      541 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
      601 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
      661 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
      721 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
      781 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
      841 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
      901 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
      961 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     1021 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     1081 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     1141 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     1201 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     1261 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     1321 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     1381 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     1441 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     1501 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     1561 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     1621 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     1681 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     1741 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgggaa
     1801 attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     1861 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     1921 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     1981 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     2041 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     2101 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     2161 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     2221 cccgccgcgc ttaatgcgcc gctacagggc gcgtcgcgcc attcgccatt caggctgcgc
     2281 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     2341 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     2401 aaaacgacgg ccagtgaatt gtaatacgac tcactatagg gcgaattgga gctcggtatt
     2461 cacgacagca ggctgaataa taaaaaaatt agaaactatt atttaaccct agaaagataa
     2521 tcatattgtg acgtacgtta aagataatca tgcgtaaaat tgacgcatgt gttttatcgg
     2581 tctgtatatc gaggtttatt tattaatttg aatagatatt aagttttatt atatttacac
     2641 ttacatacta ataataaatt caacaaacaa tttatttatg tttatttatt tattaaaaaa
     2701 aaacaaaaac tcaaaatttc ttctataaag taacaaaact tttaaacatt ctctctttta
     2761 caaaaataaa cttattttgt actttaaaaa cagtcatgtt gtattataaa ataagtaatt
     2821 agcttaacct atacataata gaaacaaatt atacttatta gtcagtcaga aacaactttg
     2881 gcacatatca atattatgct ctcgttaatt aactcgaggg atccaagctg gccgcacgtc
     2941 tccctatcag tgatagagaa gtcgacacgt ctcgagctcc ctatcagtga tagagaaggt
     3001 acgtctagaa cgtctcccta tcagtgatag agaagtcgac acgtctcgag ctccctatca
     3061 gtgatagaga aggtacgtct agaacgtctc cctatcagtg atagagaagt cgacacgtct
     3121 cgagctccct atcagtgata gagaaggtac gtctagaacg tctccctatc agtgatagag
     3181 aagtcgacac gtctcgagct ccctatcagt gatagagaag gtacccccta tataagcaga
     3241 gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat
     3301 agaagacacc gggaccgatc cagcctccgc ggtggcggcc atcacaagtt tgactagtac
     3361 cggttactcg aggctagcac catggcagaa atcggtactg gctttccatt cgacccccat
     3421 tatgtggaag tcctgggcga gcgcatgcac tacgtcgatg ttggtccgcg cgatggcacc
     3481 cctgtgctgt tcctgcacgg taacccgacc tcctcctacg tgtggcgcaa catcatcccg
     3541 catgttgcac cgacccatcg ctgcattgct ccagacctga tcggtatggg caaatccgac
     3601 aaaccagacc tgggttattt cttcgacgac cacgtccgct tcatggatgc cttcatcgaa
     3661 gccctgggtc tggaagaggt cgtcctggtc attcacgact ggggctccgc tctgggtttc
     3721 cactgggcca agcgcaatcc agagcgcgtc aaaggtattg catttatgga gttcatccgc
     3781 cctatcccga cctgggacga atggccagaa tttgcccgcg agaccttcca ggccttccgc
     3841 accaccgacg tcggccgcaa gctgatcatc gatcagaacg tttttatcga gggtacgctg
     3901 ccgatgggtg tcgtccgccc gctgactgaa gtcgagatgg accattaccg cgagccgttc
     3961 ctgaatcctg ttgaccgcga gccactgtgg cgcttcccaa acgagctgcc aatcgccggt
     4021 gagccagcga acatcgtcgc gctggtcgaa gaatacatgg actggctgca ccagtcccct
     4081 gtcccgaagc tgctgttctg gggcacccca ggcgttctga tcccaccggc cgaagccgct
     4141 cgcctggcca aaagcctgcc taactgcaag gctgtggaca tcggcccggg tctgaatctg
     4201 ctgcaagaag acaacccgga cctgatcggc agcgagatcg cgcgctggct gtcgacgctc
     4261 gagatttccg gcgagccaac cactgaggat ctgtactttc agagcgataa cgcgatcgcc
     4321 ggcgcgccaa caagtttgta caaaaaagct gaacgagaaa cgtaaaatga tataaatatc
     4381 aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat
     4441 atccagtcac tatggcggcc gcattaggca ccccaggctt tacactttat gcttccggct
     4501 cgtataatgt gtggattttg agttaggatc cgtcgagatt ttcaggagct aaggaagcta
     4561 aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     4621 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     4681 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     4741 ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg
     4801 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     4861 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     4921 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     4981 agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg
     5041 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     5101 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     5161 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     5221 aaagatctgg atcccgacaa tcaacgcgtg gatccggctt actaaaagcc agataacagt
     5281 atgcgtattt gcgcgctgat ttttgcggta taagaatata tactgatatg tatacccgaa
     5341 gtatgtcaaa aagaggtatg ctatgaagca gcgtattaca gtgacagttg acagcgacag
     5401 ctatcagttg ctcaaggcat atatgatgtc aatatctccg gtctggtaag cacaaccatg
     5461 cagaatgaag cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg
     5521 gctgaggtcg cccggtttat tgaaatgaac ggctcttttg ctgacgagaa caggggctgg
     5581 tgaaatgcag tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga
     5641 tgtacagagt gatattattg acacgcccgg gcgacggatg gtgatccccc tggccagtgc
     5701 acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga
     5761 aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta tcggggaaga
     5821 agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg
     5881 gggaatataa atgtcaggct cccttataca cagccagtct gcaggtcgac catagtgact
     5941 ggatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata
     6001 tattgatatt tatatcattt tacgtttctc gttcagcttt cttgtacaaa gtggtgatct
     6061 agctaaaacg cggcctcgaa tcgaattccg cccccccccc tctccctccc ccccccctaa
     6121 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     6181 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     6241 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     6301 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     6361 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     6421 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     6481 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     6541 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
     6601 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     6661 acgatgataa tatggccaca accatggcca caaccatgga cgggccgcgc ctgctgctgt
     6721 tgctgcttct gggggtgtcc cttggaggtg ccaaggaggc atgccccaca ggcctgtaca
     6781 cacacagcgg tgagtgctgc aaagcctgca acctgggcga gggtgtggcc cagccttgtg
     6841 gagccaacca gaccgtgtgt gagccctgcc tggacagcgt gacgttctcc gacgtggtga
     6901 gcgcgaccga gccgtgcaag ccgtgcaccg agtgcgtggg gctccagagc atgtcggcgc
     6961 cgtgcgtgga ggccgacgac gccgtgtgcc gctgcgccta cggctactac caggatgaga
     7021 cgactgggcg ctgcgaggcg tgccgcgtgt gcgaggcggg ctcgggcctc gtgttctcct
     7081 gccaggacaa gcagaacacc gtgtgcgagg agtgccccga cggcacgtat tccgacgagg
     7141 ccaaccacgt ggacccgtgc ctgccctgca ccgtgtgcga ggacaccgag cgccagctcc
     7201 gcgagtgcac acgctgggcc gacgccgagt gcgaggagat ccctggccgt tggattacac
     7261 ggtccacacc cccagagggc tcggacagca cagcccccag cacccaggag cctgaggcac
     7321 ctccagaaca agacctcata gccagcacgg tggcaggtgt ggtgaccaca gtgatgggca
     7381 gctcccagcc cgtggtgacc cgaggcacca ccgacaacct catccctgtc tattgctcca
     7441 tcctggctgc tgtggttgtg ggccttgtgg cctacatagc cttcaagagg tggaacagtc
     7501 atcgatatcc tcgaggtcac cgcggtctag agtcgacctg cagccaagct tatcgataaa
     7561 gcggccgcga tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc
     7621 atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     7681 tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt
     7741 atttggttta gagtttggca acatatgcca tatgctggct gccatgaaca aaggtggcta
     7801 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     7861 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     7921 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     7981 ccagtcatag ctgtccctct tctcttatga agatccctcg acctgcagga attcgattac
     8041 ttaagcctag gatccagatc tgtacattcg aatcgatgaa ttcgttaacc atggaagctt
     8101 ctgcagcggc cgcactagcg gtggcggcct cgacattgat tattgactag ttattaatag
     8161 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     8221 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     8281 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     8341 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     8401 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     8461 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat gatgatgcgg
     8521 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     8581 caccccattg acgtcaatgg gagtttgttt tgactagtgg agccgagagt aattcataca
     8641 aaaggaggga tcgccttcgc aaggggagag cccagggacc gtccctaaat tctcacagac
     8701 ccaaatccct gtagccgccc cacgacagcg cgaggagcat gcgcccaggg ctgagcgcgg
     8761 gtagatcaga gcacacaagc tcacagtccc cggcggtggg gggaggggcg cgctgagcgg
     8821 gggccaggga gctggcgcgg ggcaaactgg gaaagtggtg tcgtgtgctg gctccgccct
     8881 cttcccgagg gtgggggaga acggtatata agtgcggtag tcgccttgga cgttcttttt
     8941 cgcaacgggt ttgccgtcag aacgcaggtg agtggcgggt gtggcttccg cgggccccgg
     9001 agctggagcc ctgctctgag cgggccgggc tgatatgcga gtgtcgtccg cagggtttag
     9061 ctgtgagcat tcccacttcg agtggcgggc ggtgcggggg tgagagtgcg aggcctagcg
     9121 gcaaccccgt agcctcgcct cgtgtccggc ttgaggccta gcgtggtgtc cgccgccgcg
     9181 tgccactccg gccgcactat gcgttttttg tccttgctgc cctcgattgc cttccagcag
     9241 catgggctaa caaagggagg gtgtggggct cactcttaag gagcccatga agcttacgtt
     9301 ggataggaat ggaagggcag gaggggcgac tggggcccgc ccgccttcgg agcacatgtc
     9361 cgacgccacc tggatggggc gaggcctgtg gctttccgaa gcaatcgggc gtgagtttag
     9421 cctacctggg ccatgtggcc ctagcactgg gcacggtctg gcctggcggt gccgcgttcc
     9481 cttgcctccc aacaagggtg aggccgtccc gcccggcacc agttgcttgc gcggaaagat
     9541 ggccgctccc ggggccctgt tgcaaggagc tcaaaatgga ggacgcggca gcccggtgga
     9601 gcgggcgggt gagtcaccca cacaaaggaa gagggccttg cccctcgccg gccgctgctt
     9661 cctgtgaccc cgtggtctat cggccgcata gtcacctcgg gcttctcttg agcaccgctc
     9721 gtcgcggcgg ggggagggga tctaatggcg ttggagtttg ttcacatttg gtgggtggag
     9781 actagtcagg ccagcctggc gctggaagtc attcttggaa tttgcccctt tgagtttgga
     9841 gcgaggctaa ttctcaagcc tcttagcggt tcaaaggtat tttctaaacc cgtttccagg
     9901 tgttgtgaaa gccaccgcta attcaaagca atccggagta aattcaccat gtctagactg
     9961 gacaagagca aagtcataaa cggcgctctg gaattactca atggagtcgg tatcgaaggc
    10021 ctgacgacaa ggaaactcgc tcaaaagctg ggagttgagc agcctaccct gtactggcac
    10081 gtgaagaaca agcgggccct gctcgatgcc ctgccaatcg agatgctgga caggcatcat
    10141 acccacttct gccccctgga aggcgagtca tggcaagact ttctgcggaa caacgccaag
    10201 tcattccgct gtgctctcct ctcacatcgc gacggggcta aagtgcatct cggcacccgc
    10261 ccaacagaga aacagtacga aaccctggaa aatcagctcg cgttcctgtg tcagcaaggc
    10321 ttctccctgg agaacgcact gtacgctctg tccgccgtgg gccactttac actgggctgc
    10381 gtattggagg aacaggagca tcaagtagca aaagaggaaa gagagacacc taccaccgat
    10441 tctatgcccc cacttctgag acaagcaatt gagctgttcg accggcaggg agccgaacct
    10501 gccttccttt tcggcctgga actaatcata tgtggcctgg agaaacagct aaagtgcgaa
    10561 agcggcgggc cggccgacgc ccttgacgat tttgacttag acatgctccc agccgatgcc
    10621 cttgacgact ttgaccttga tatgctgcct gctgacgctc ttgacgattt tgaccttgac
    10681 atgctccccg ggtaactaag taaggatccc gtacgcctag gagcaggttt ccccaatgac
    10741 acaaaacgtg caacttgaaa ctccgcctgg tctttccagg tctagagggg taacactttg
    10801 tactgcgttt ggctccacgc tcgatccact ggcgagtgtt agtaacagca ctgttgcttc
    10861 gtagcggagc atgacggccg tgggaactcc tccttggtaa caaggaccca cggggccaaa
    10921 agccacgccc acacgggccc gtcatgtgtg caaccccagc acggcgactt tactgcgaaa
    10981 cccactttaa agtgacattg aaactggtac ccacacactg gtgacaggct aaggatgccc
    11041 ttcaggtacc ccgaggtaac acgcgacact cgggatctga gaaggggact ggggcttcta
    11101 taaaagcgct cggtttaaaa agcttctatg cctgaatagg tgaccggagg tcggcacctt
    11161 tcctttgcaa ttactgaccc tatgaataca ctgactgttt gacaattaat catcggcata
    11221 gtatatcggc atagtataat acgactcact ataggagggc caccatgatt gaacaagatg
    11281 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
    11341 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
    11401 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
    11461 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
    11521 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
    11581 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
    11641 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
    11701 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
    11761 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
    11821 tgacacatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
    11881 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
    11941 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
    12001 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
    12061 cgggactctg gggttcgaaa tgaccgacca agcgaattcg ctaggattat ccctaatacc
    12121 tgccacccca ctcttaatca gtggtggaag aacggtctca gaactgtttg tttcaattgg
    12181 ccatttaagt ttagtagtaa aagactggtt aatgataaca atgcatcgta aaaccttcag
    12241 aaggaaagga gaatgttttg tggaccactt tggttttctt ttttgcgtgt ggcagtttta
    12301 agttattagt ttttaaaatc agtacttttt aatggaaaca acttgaccaa aaatttgtca
    12361 cagaattttg agacccatta aaaaagttaa atgagaaacc tgtgtgttcc tttggtcaac
    12421 accgagacat ttaggtgaaa gacatctaat tctggtttta cgaatctgga aacttcttga
    12481 aaatgtaatt cttgagttaa cacttctggg tggagaatag ggttgttttc cccccacata
    12541 attggaaggg gaaggaatat catttaaagc tatgggaggg ttgctttgat tacaacactg
    12601 gagagaaatg cagcatgttg ctgattgcct gtcactaaaa caggccaaaa actgagtcct
    12661 tgggttgcat agaaagctgc caaacgctag ccaacaagct cgtcatcgct ttgcagaaga
    12721 gcagagagga tatgctcatc gtctaaagaa ctacccattt tattatatat tagtcacgat
    12781 atctataaca agaaaatata tatataataa gttatcacgt aagtagaaca tgaaataaca
    12841 atataattat cgtatgagtt aaatcttaaa agtcacgtaa aagataatca tgcgtcattt
    12901 tgactcacgc ggtcgttata gttcaaaatc agtgacactt accgcattga caagcacgcc
    12961 tcacgggagc tccaagcggc gactgagatg tcctaaatgc acagcgacgg attcgcgcta
    13021 tttagaaaga gagagcaata tttcaagaat gcatgcgtca attttacgca gactatcttt
    13081 ctagggttaa aaaagatttg cgctttactc gacctaaact ttaaacacgt catagaatct
    13141 tcgtttgaca aaaaccacat tgtggccaag ctgtgtgacg cgacgcgcgc taaagaatgg
    13201 caaaccaagt cgcgcgaggt acccagcttt tgttcccttt agtgagggtt aattccgagc
    13261 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
    13321 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
    13381 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
    13441 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
    13501 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
    13561 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
    13621 tgagcaaaag g
//
