LOCUS       sequence-324912-       13625 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 13625)
  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 13625)
  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..13625
                     /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(1796..2251)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1933..1954)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2145..2164
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2378..2400
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2392..2409
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2393..2409
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2416..2434
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   2501..2563
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    2935..2953
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2972..2990
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3009..3027
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3046..3064
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3083..3101
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3120..3138
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3157..3175
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3194..3212
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3234..3258
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3376..4266
                     /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             4279..4299
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     protein_bind    4324..4448
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(4473..4478,4479..4496,4497..4503)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             4557..5216
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4623..4642)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             5579..5884
                     /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     5763..5782
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(5925..6049)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    join(6091..6665,6666..6668,6669..6677)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6258..6275)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6485..6504
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(7565..7584)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7630..7685
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7684..7703)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     enhancer        8128..8507
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8622..9927
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    8964..9895
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     CDS             join(9944..10561,10562..10567,10568..10603,10604..10606,
                     10607..10642,10643..10645,10646..10681,10682..10690)
                     /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    10717..11161
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        11194..11240
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        11232..11250
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             11260..12054
                     /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             11260..12054
                     /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(11314..11333)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     11924..11943
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    12101..12673
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     repeat_region   13047..13081
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(13230..13248)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(13269..13285)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13269..13285)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13282..13304)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13293..13309
                     /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(13317..13323,13324..13341,13342..13347))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13362..13383
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13500..13517)
                     /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 tagaagaaca 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 cacctaaatt
     1801 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     1861 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     1921 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     1981 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2041 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2101 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2161 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     2221 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     2281 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     2341 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     2401 acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcgg tattcacgac
     2461 agcaggctga ataataaaaa aattagaaac tattatttaa ccctagaaag ataatcatat
     2521 tgtgacgtac gttaaagata atcatgcgta aaattgacgc atgtgtttta tcggtctgta
     2581 tatcgaggtt tatttattaa tttgaataga tattaagttt tattatattt acacttacat
     2641 actaataata aattcaacaa acaatttatt tatgtttatt tatttattaa aaaaaaacaa
     2701 aaactcaaaa tttcttctat aaagtaacaa aacttttaaa cattctctct tttacaaaaa
     2761 taaacttatt ttgtacttta aaaacagtca tgttgtatta taaaataagt aattagctta
     2821 acctatacat aatagaaaca aattatactt attagtcagt cagaaacaac tttggcacat
     2881 atcaatatta tgctctcgtt aattaactcg agggatccaa gctggccgca cgtctcccta
     2941 tcagtgatag agaagtcgac acgtctcgag ctccctatca gtgatagaga aggtacgtct
     3001 agaacgtctc cctatcagtg atagagaagt cgacacgtct cgagctccct atcagtgata
     3061 gagaaggtac gtctagaacg tctccctatc agtgatagag aagtcgacac gtctcgagct
     3121 ccctatcagt gatagagaag gtacgtctag aacgtctccc tatcagtgat agagaagtcg
     3181 acacgtctcg agctccctat cagtgataga gaaggtaccc cctatataag cagagctcgt
     3241 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
     3301 caccgggacc gatccagcct ccgcggtggc ggccatcaca agtttgacta gtaccggtta
     3361 ctcgaggcta gcaccatggc agaaatcggt actggctttc cattcgaccc ccattatgtg
     3421 gaagtcctgg gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg cacccctgtg
     3481 ctgttcctgc acggtaaccc gacctcctcc tacgtgtggc gcaacatcat cccgcatgtt
     3541 gcaccgaccc atcgctgcat tgctccagac ctgatcggta tgggcaaatc cgacaaacca
     3601 gacctgggtt atttcttcga cgaccacgtc cgcttcatgg atgccttcat cgaagccctg
     3661 ggtctggaag aggtcgtcct ggtcattcac gactggggct ccgctctggg tttccactgg
     3721 gccaagcgca atccagagcg cgtcaaaggt attgcattta tggagttcat ccgccctatc
     3781 ccgacctggg acgaatggcc agaatttgcc cgcgagacct tccaggcctt ccgcaccacc
     3841 gacgtcggcc gcaagctgat catcgatcag aacgttttta tcgagggtac gctgccgatg
     3901 ggtgtcgtcc gcccgctgac tgaagtcgag atggaccatt accgcgagcc gttcctgaat
     3961 cctgttgacc gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca
     4021 gcgaacatcg tcgcgctggt cgaagaatac atggactggc tgcaccagtc ccctgtcccg
     4081 aagctgctgt tctggggcac cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg
     4141 gccaaaagcc tgcctaactg caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa
     4201 gaagacaacc cggacctgat cggcagcgag atcgcgcgct ggctgtcgac gctcgagatt
     4261 tccggcgagc caaccactga ggatctgtac tttcagagcg ataacgcgat cgccggcgcg
     4321 ccaacaagtt tgtacaaaaa agctgaacga gaaacgtaaa atgatataaa tatcaatata
     4381 ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag
     4441 tcactatggc ggccgcatta ggcaccccag gctttacact ttatgcttcc ggctcgtata
     4501 atgtgtggat tttgagttag gatccgtcga gattttcagg agctaaggaa gctaaaatgg
     4561 agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt
     4621 ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta
     4681 cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca
     4741 ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa gacggtgagc
     4801 tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt
     4861 tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc
     4921 aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata
     4981 tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca
     5041 atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca
     5101 aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg
     5161 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaaagat
     5221 ctggatcccg acaatcaacg cgtggatccg gcttactaaa agccagataa cagtatgcgt
     5281 atttgcgcgc tgatttttgc ggtataagaa tatatactga tatgtatacc cgaagtatgt
     5341 caaaaagagg tatgctatga agcagcgtat tacagtgaca gttgacagcg acagctatca
     5401 gttgctcaag gcatatatga tgtcaatatc tccggtctgg taagcacaac catgcagaat
     5461 gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg gatggctgag
     5521 gtcgcccggt ttattgaaat gaacggctct tttgctgacg agaacagggg ctggtgaaat
     5581 gcagtttaag gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca
     5641 gagtgatatt attgacacgc ccgggcgacg gatggtgatc cccctggcca gtgcacgtct
     5701 gctgtcagat aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg
     5761 gcgcatgatg accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc
     5821 tgatctcagc caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat
     5881 ataaatgtca ggctccctta tacacagcca gtctgcaggt cgaccatagt gactggatat
     5941 gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga
     6001 tatttatatc attttacgtt tctcgttcag ctttcttgta caaagtggtg atctagctaa
     6061 aacgcggcct cgaatcgaat tccgcccccc cccctctccc tccccccccc ctaacgttac
     6121 tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat
     6181 attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat
     6241 tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga
     6301 agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca
     6361 gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac
     6421 acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt
     6481 caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca
     6541 ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt
     6601 aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg
     6661 ataatatggc cacaaccatg gccacaacca tggacgggcc gcgcctgctg ctgttgctgc
     6721 ttctgggggt gtcccttgga ggtgccaagg aggcatgccc cacaggcctg tacacacaca
     6781 gcggtgagtg ctgcaaagcc tgcaacctgg gcgagggtgt ggcccagcct tgtggagcca
     6841 accagaccgt gtgtgagccc tgcctggaca gcgtgacgtt ctccgacgtg gtgagcgcga
     6901 ccgagccgtg caagccgtgc accgagtgcg tggggctcca gagcatgtcg gcgccgtgcg
     6961 tggaggccga cgacgccgtg tgccgctgcg cctacggcta ctaccaggat gagacgactg
     7021 ggcgctgcga ggcgtgccgc gtgtgcgagg cgggctcggg cctcgtgttc tcctgccagg
     7081 acaagcagaa caccgtgtgc gaggagtgcc ccgacggcac gtattccgac gaggccaacc
     7141 acgtggaccc gtgcctgccc tgcaccgtgt gcgaggacac cgagcgccag ctccgcgagt
     7201 gcacacgctg ggccgacgcc gagtgcgagg agatccctgg ccgttggatt acacggtcca
     7261 cacccccaga gggctcggac agcacagccc ccagcaccca ggagcctgag gcacctccag
     7321 aacaagacct catagccagc acggtggcag gtgtggtgac cacagtgatg ggcagctccc
     7381 agcccgtggt gacccgaggc accaccgaca acctcatccc tgtctattgc tccatcctgg
     7441 ctgctgtggt tgtgggcctt gtggcctaca tagccttcaa gaggtggaac agtcatcgat
     7501 atcctcgagg tcaccgcggt ctagagtcga cctgcagcca agcttatcga taaagcggcc
     7561 gcgatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga
     7621 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     7681 tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg
     7741 tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa
     7801 gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct
     7861 tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc
     7921 taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag
     7981 tcatagctgt ccctcttctc ttatggagat ccctcgacct gcaggaattc gattacttaa
     8041 gcctaggatc cagatctgta cattcgaatc gatgaattcg ttaaccatgg aagcttctgc
     8101 agcggccgca ctagcggtgg cggcctcgac attgattatt gactagttat taatagtaat
     8161 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     8221 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     8281 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     8341 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     8401 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     8461 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgatg atgcggtttt
     8521 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     8581 ccattgacgt caatgggagt ttgttttgac tagtggagcc gagagtaatt catacaaaag
     8641 gagggatcgc cttcgcaagg ggagagccca gggaccgtcc ctaaattctc acagacccaa
     8701 atccctgtag ccgccccacg acagcgcgag gagcatgcgc ccagggctga gcgcgggtag
     8761 atcagagcac acaagctcac agtccccggc ggtgggggga ggggcgcgct gagcgggggc
     8821 cagggagctg gcgcggggca aactgggaaa gtggtgtcgt gtgctggctc cgccctcttc
     8881 ccgagggtgg gggagaacgg tatataagtg cggtagtcgc cttggacgtt ctttttcgca
     8941 acgggtttgc cgtcagaacg caggtgagtg gcgggtgtgg cttccgcggg ccccggagct
     9001 ggagccctgc tctgagcggg ccgggctgat atgcgagtgt cgtccgcagg gtttagctgt
     9061 gagcattccc acttcgagtg gcgggcggtg cgggggtgag agtgcgaggc ctagcggcaa
     9121 ccccgtagcc tcgcctcgtg tccggcttga ggcctagcgt ggtgtccgcc gccgcgtgcc
     9181 actccggccg cactatgcgt tttttgtcct tgctgccctc gattgccttc cagcagcatg
     9241 ggctaacaaa gggagggtgt ggggctcact cttaaggagc ccatgaagct tacgttggat
     9301 aggaatggaa gggcaggagg ggcgactggg gcccgcccgc cttcggagca catgtccgac
     9361 gccacctgga tggggcgagg cctgtggctt tccgaagcaa tcgggcgtga gtttagccta
     9421 cctgggccat gtggccctag cactgggcac ggtctggcct ggcggtgccg cgttcccttg
     9481 cctcccaaca agggtgaggc cgtcccgccc ggcaccagtt gcttgcgcgg aaagatggcc
     9541 gctcccgggg ccctgttgca aggagctcaa aatggaggac gcggcagccc ggtggagcgg
     9601 gcgggtgagt cacccacaca aaggaagagg gccttgcccc tcgccggccg ctgcttcctg
     9661 tgaccccgtg gtctatcggc cgcatagtca cctcgggctt ctcttgagca ccgctcgtcg
     9721 cggcgggggg aggggatcta atggcgttgg agtttgttca catttggtgg gtggagacta
     9781 gtcaggccag cctggcgctg gaagtcattc ttggaatttg cccctttgag tttggagcga
     9841 ggctaattct caagcctctt agcggttcaa aggtattttc taaacccgtt tccaggtgtt
     9901 gtgaaagcca ccgctaattc aaagcaatcc ggagtaattc accatgtcta gactggacaa
     9961 gagcaaagtc ataaacggcg ctctggaatt actcaatgga gtcggtatcg aaggcctgac
    10021 gacaaggaaa ctcgctcaaa agctgggagt tgagcagcct accctgtact ggcacgtgaa
    10081 gaacaagcgg gccctgctcg atgccctgcc aatcgagatg ctggacaggc atcataccca
    10141 cttctgcccc ctggaaggcg agtcatggca agactttctg cggaacaacg ccaagtcatt
    10201 ccgctgtgct ctcctctcac atcgcgacgg ggctaaagtg catctcggca cccgcccaac
    10261 agagaaacag tacgaaaccc tggaaaatca gctcgcgttc ctgtgtcagc aaggcttctc
    10321 cctggagaac gcactgtacg ctctgtccgc cgtgggccac tttacactgg gctgcgtatt
    10381 ggaggaacag gagcatcaag tagcaaaaga ggaaagagag acacctacca ccgattctat
    10441 gcccccactt ctgagacaag caattgagct gttcgaccgg cagggagccg aacctgcctt
    10501 ccttttcggc ctggaactaa tcatatgtgg cctggagaaa cagctaaagt gcgaaagcgg
    10561 cgggccggcc gacgcccttg acgattttga cttagacatg ctcccagccg atgcccttga
    10621 cgactttgac cttgatatgc tgcctgctga cgctcttgac gattttgacc ttgacatgct
    10681 ccccgggtaa ctaagtaagg atcccgtacg cctaggagca ggtttcccca atgacacaaa
    10741 acgtgcaact tgaaactccg cctggtcttt ccaggtctag aggggtaaca ctttgtactg
    10801 cgtttggctc cacgctcgat ccactggcga gtgttagtaa cagcactgtt gcttcgtagc
    10861 ggagcatgac ggccgtggga actcctcctt ggtaacaagg acccacgggg ccaaaagcca
    10921 cgcccacacg ggcccgtcat gtgtgcaacc ccagcacggc gactttactg cgaaacccac
    10981 tttaaagtga cattgaaact ggtacccaca cactggtgac aggctaagga tgcccttcag
    11041 gtaccccgag gtaacacgcg acactcggga tctgagaagg ggactggggc ttctataaaa
    11101 gcgctcggtt taaaaagctt ctatgcctga ataggtgacc ggaggtcggc acctttcctt
    11161 tgcaattact gaccctatga atacactgac tgtttgacaa ttaatcatcg gcatagtata
    11221 tcggcatagt ataatacgac tcactatagg agggccacca tgattgaaca agatggattg
    11281 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
    11341 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
    11401 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
    11461 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
    11521 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
    11581 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
    11641 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
    11701 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
    11761 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgaca
    11821 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
    11881 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
    11941 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
    12001 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
    12061 ctctggggtt cgaaatgacc gaccaagcga attcgctagg attatcccta atacctgcca
    12121 ccccactctt aatcagtggt ggaagaacgg tctcagaact gtttgtttca attggccatt
    12181 taagtttagt agtaaaagac tggttaatga taacaatgca tcgtaaaacc ttcagaagga
    12241 aaggagaatg ttttgtggac cactttggtt ttcttttttg cgtgtggcag ttttaagtta
    12301 ttagttttta aaatcagtac tttttaatgg aaacaacttg accaaaaatt tgtcacagaa
    12361 ttttgagacc cattaaaaaa gttaaatgag aaacctgtgt gttcctttgg tcaacaccga
    12421 gacatttagg tgaaagacat ctaattctgg ttttacgaat ctggaaactt cttgaaaatg
    12481 taattcttga gttaacactt ctgggtggag aatagggttg ttttcccccc acataattgg
    12541 aaggggaagg aatatcattt aaagctatgg gagggttgct ttgattacaa cactggagag
    12601 aaatgcagca tgttgctgat tgcctgtcac taaaacaggc caaaaactga gtccttgggt
    12661 tgcatagaaa gctgccaaac gctagccaac aagctcgtca tcgctttgca gaagagcaga
    12721 gaggatatgc tcatcgtcta aagaactacc cattttatta tatattagtc acgatatcta
    12781 taacaagaaa atatatatat aataagttat cacgtaagta gaacatgaaa taacaatata
    12841 attatcgtat gagttaaatc ttaaaagtca cgtaaaagat aatcatgcgt cattttgact
    12901 cacgcggtcg ttatagttca aaatcagtga cacttaccgc attgacaagc acgcctcacg
    12961 ggagctccaa gcggcgactg agatgtccta aatgcacagc gacggattcg cgctatttag
    13021 aaagagagag caatatttca agaatgcatg cgtcaatttt acgcagacta tctttctagg
    13081 gttaaaaaag atttgcgctt tactcgacct aaactttaaa cacgtcatag aatcttcgtt
    13141 tgacaaaaac cacattgtgg ccaagctgtg tgacgcgacg cgcgctaaag aatggcaaac
    13201 caagtcgcgc gaggtaccag cttttgttcc ctttagtgag ggttaatttc gagcttggcg
    13261 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
    13321 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
    13381 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
    13441 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
    13501 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
    13561 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
    13621 aaagg
//
