LOCUS       sequence-324198-       13484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  All-in-One piggyBac transposon Gateway Destination vector for 
            dox-inducible expression of N-terminal His-TEV-GFP 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 13484)
  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 13484)
  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-6HIS-TEV-mEGFP,IRES-NGFR
FEATURES             Location/Qualifiers
     source          1..13484
                     /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             3388..3405
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3412..3432
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             join(3442..3444,3445..3447,3448..4158)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3487..3508)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3748..3767)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4095..4116
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    4183..4307
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(4332..4337,4338..4355,4356..4362)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             4416..5075
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4482..4501)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             5438..5743
                     /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     5622..5641
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(5784..5908)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    join(5950..6524,6525..6527,6528..6536)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6117..6134)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6344..6363
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(7424..7443)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7489..7544
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7543..7562)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     enhancer        7987..8366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8481..9786
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    8823..9754
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     CDS             join(9803..10420,10421..10426,10427..10462,10463..10465,
                     10466..10501,10502..10504,10505..10540,10541..10549)
                     /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    10576..11020
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        11053..11099
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        11091..11109
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             11119..11913
                     /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             11119..11913
                     /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(11173..11192)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     11783..11802
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    11960..12532
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     repeat_region   12906..12940
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(13089..13107)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(13128..13144)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13128..13144)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13141..13163)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13152..13168
                     /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(13176..13182,13183..13200,13201..13206))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13221..13242
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13359..13376)
                     /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 gcaccatggg cagcagccat catcatcatc atcacagcag cgaggatctg
     3421 tactttcaga gcggcagcag catggtgagc aagggcgagg agctgttcac cggggtggtg
     3481 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     3541 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     3601 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     3661 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     3721 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     3781 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     3841 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     3901 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     3961 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     4021 gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac
     4081 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     4141 atggacgagc tgtacaagga taacgcgatc gccggcgcgc caacaagttt gtacaaaaaa
     4201 gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa
     4261 aaacagacta cataatactg taaaacacaa catatccagt cactatggcg gccgcattag
     4321 gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt ttgagttagg
     4381 atccgtcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata
     4441 ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg
     4501 ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa
     4561 agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg
     4621 ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc
     4681 acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat
     4741 accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg
     4801 aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc
     4861 cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc
     4921 ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga
     4981 ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac
     5041 aacagtactg cgatgagtgg cagggcgggg cgtaaagatc tggatcccga caatcaacgc
     5101 gtggatccgg cttactaaaa gccagataac agtatgcgta tttgcgcgct gatttttgcg
     5161 gtataagaat atatactgat atgtataccc gaagtatgtc aaaaagaggt atgctatgaa
     5221 gcagcgtatt acagtgacag ttgacagcga cagctatcag ttgctcaagg catatatgat
     5281 gtcaatatct ccggtctggt aagcacaacc atgcagaatg aagcccgtcg tctgcgtgcc
     5341 gaacgctgga aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt tattgaaatg
     5401 aacggctctt ttgctgacga gaacaggggc tggtgaaatg cagtttaagg tttacaccta
     5461 taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc
     5521 cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtctcccg
     5581 tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat
     5641 ggccagtgtg ccggtctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa
     5701 tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag gctcccttat
     5761 acacagccag tctgcaggtc gaccatagtg actggatatg ttgtgtttta cagtattatg
     5821 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
     5881 ctcgttcagc tttcttgtac aaagtggtga tctagctaaa acgcggcctc gaatcgaatt
     5941 ccgccccccc ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata
     6001 aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt
     6061 gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct
     6121 cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc
     6181 ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga
     6241 caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc
     6301 ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt
     6361 attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg
     6421 gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc
     6481 gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc acaaccatgg
     6541 ccacaaccat ggacgggccg cgcctgctgc tgttgctgct tctgggggtg tcccttggag
     6601 gtgccaagga ggcatgcccc acaggcctgt acacacacag cggtgagtgc tgcaaagcct
     6661 gcaacctggg cgagggtgtg gcccagcctt gtggagccaa ccagaccgtg tgtgagccct
     6721 gcctggacag cgtgacgttc tccgacgtgg tgagcgcgac cgagccgtgc aagccgtgca
     6781 ccgagtgcgt ggggctccag agcatgtcgg cgccgtgcgt ggaggccgac gacgccgtgt
     6841 gccgctgcgc ctacggctac taccaggatg agacgactgg gcgctgcgag gcgtgccgcg
     6901 tgtgcgaggc gggctcgggc ctcgtgttct cctgccagga caagcagaac accgtgtgcg
     6961 aggagtgccc cgacggcacg tattccgacg aggccaacca cgtggacccg tgcctgccct
     7021 gcaccgtgtg cgaggacacc gagcgccagc tccgcgagtg cacacgctgg gccgacgccg
     7081 agtgcgagga gatccctggc cgttggatta cacggtccac acccccagag ggctcggaca
     7141 gcacagcccc cagcacccag gagcctgagg cacctccaga acaagacctc atagccagca
     7201 cggtggcagg tgtggtgacc acagtgatgg gcagctccca gcccgtggtg acccgaggca
     7261 ccaccgacaa cctcatccct gtctattgct ccatcctggc tgctgtggtt gtgggccttg
     7321 tggcctacat agccttcaag aggtggaaca gtcatcgata tcctcgaggt caccgcggtc
     7381 tagagtcgac ctgcagccaa gcttatcgat aaagcggccg cgatcttttt ccctctgcca
     7441 aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat
     7501 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     7561 ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg
     7621 cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac
     7681 agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt
     7741 ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt
     7801 tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct
     7861 tatggagatc cctcgacctg caggaattcg attacttaag cctaggatcc agatctgtac
     7921 attcgaatcg atgaattcgt taaccatgga agcttctgca gcggccgcac tagcggtggc
     7981 ggcctcgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     8041 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     8101 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     8161 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     8221 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     8281 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     8341 acgtattagt catcgctatt accatgatga tgcggttttg gcagtacatc aatgggcgtg
     8401 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     8461 tgttttgact agtggagccg agagtaattc atacaaaagg agggatcgcc ttcgcaaggg
     8521 gagagcccag ggaccgtccc taaattctca cagacccaaa tccctgtagc cgccccacga
     8581 cagcgcgagg agcatgcgcc cagggctgag cgcgggtaga tcagagcaca caagctcaca
     8641 gtccccggcg gtggggggag gggcgcgctg agcgggggcc agggagctgg cgcggggcaa
     8701 actgggaaag tggtgtcgtg tgctggctcc gccctcttcc cgagggtggg ggagaacggt
     8761 atataagtgc ggtagtcgcc ttggacgttc tttttcgcaa cgggtttgcc gtcagaacgc
     8821 aggtgagtgg cgggtgtggc ttccgcgggc cccggagctg gagccctgct ctgagcgggc
     8881 cgggctgata tgcgagtgtc gtccgcaggg tttagctgtg agcattccca cttcgagtgg
     8941 cgggcggtgc gggggtgaga gtgcgaggcc tagcggcaac cccgtagcct cgcctcgtgt
     9001 ccggcttgag gcctagcgtg gtgtccgccg ccgcgtgcca ctccggccgc actatgcgtt
     9061 ttttgtcctt gctgccctcg attgccttcc agcagcatgg gctaacaaag ggagggtgtg
     9121 gggctcactc ttaaggagcc catgaagctt acgttggata ggaatggaag ggcaggaggg
     9181 gcgactgggg cccgcccgcc ttcggagcac atgtccgacg ccacctggat ggggcgaggc
     9241 ctgtggcttt ccgaagcaat cgggcgtgag tttagcctac ctgggccatg tggccctagc
     9301 actgggcacg gtctggcctg gcggtgccgc gttcccttgc ctcccaacaa gggtgaggcc
     9361 gtcccgcccg gcaccagttg cttgcgcgga aagatggccg ctcccggggc cctgttgcaa
     9421 ggagctcaaa atggaggacg cggcagcccg gtggagcggg cgggtgagtc acccacacaa
     9481 aggaagaggg ccttgcccct cgccggccgc tgcttcctgt gaccccgtgg tctatcggcc
     9541 gcatagtcac ctcgggcttc tcttgagcac cgctcgtcgc ggcgggggga ggggatctaa
     9601 tggcgttgga gtttgttcac atttggtggg tggagactag tcaggccagc ctggcgctgg
     9661 aagtcattct tggaatttgc ccctttgagt ttggagcgag gctaattctc aagcctctta
     9721 gcggttcaaa ggtattttct aaacccgttt ccaggtgttg tgaaagccac cgctaattca
     9781 aagcaatccg gagtaattca ccatgtctag actggacaag agcaaagtca taaacggcgc
     9841 tctggaatta ctcaatggag tcggtatcga aggcctgacg acaaggaaac tcgctcaaaa
     9901 gctgggagtt gagcagccta ccctgtactg gcacgtgaag aacaagcggg ccctgctcga
     9961 tgccctgcca atcgagatgc tggacaggca tcatacccac ttctgccccc tggaaggcga
    10021 gtcatggcaa gactttctgc ggaacaacgc caagtcattc cgctgtgctc tcctctcaca
    10081 tcgcgacggg gctaaagtgc atctcggcac ccgcccaaca gagaaacagt acgaaaccct
    10141 ggaaaatcag ctcgcgttcc tgtgtcagca aggcttctcc ctggagaacg cactgtacgc
    10201 tctgtccgcc gtgggccact ttacactggg ctgcgtattg gaggaacagg agcatcaagt
    10261 agcaaaagag gaaagagaga cacctaccac cgattctatg cccccacttc tgagacaagc
    10321 aattgagctg ttcgaccggc agggagccga acctgccttc cttttcggcc tggaactaat
    10381 catatgtggc ctggagaaac agctaaagtg cgaaagcggc gggccggccg acgcccttga
    10441 cgattttgac ttagacatgc tcccagccga tgcccttgac gactttgacc ttgatatgct
    10501 gcctgctgac gctcttgacg attttgacct tgacatgctc cccgggtaac taagtaagga
    10561 tcccgtacgc ctaggagcag gtttccccaa tgacacaaaa cgtgcaactt gaaactccgc
    10621 ctggtctttc caggtctaga ggggtaacac tttgtactgc gtttggctcc acgctcgatc
    10681 cactggcgag tgttagtaac agcactgttg cttcgtagcg gagcatgacg gccgtgggaa
    10741 ctcctccttg gtaacaagga cccacggggc caaaagccac gcccacacgg gcccgtcatg
    10801 tgtgcaaccc cagcacggcg actttactgc gaaacccact ttaaagtgac attgaaactg
    10861 gtacccacac actggtgaca ggctaaggat gcccttcagg taccccgagg taacacgcga
    10921 cactcgggat ctgagaaggg gactggggct tctataaaag cgctcggttt aaaaagcttc
    10981 tatgcctgaa taggtgaccg gaggtcggca cctttccttt gcaattactg accctatgaa
    11041 tacactgact gtttgacaat taatcatcgg catagtatat cggcatagta taatacgact
    11101 cactatagga gggccaccat gattgaacaa gatggattgc acgcaggttc tccggccgct
    11161 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
    11221 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
    11281 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
    11341 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
    11401 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
    11461 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
    11521 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
    11581 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
    11641 aaggcgagca tgcccgacgg cgaggatctc gtcgtgacac atggcgatgc ctgcttgccg
    11701 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
    11761 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
    11821 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
    11881 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
    11941 accaagcgaa ttcgctagga ttatccctaa tacctgccac cccactctta atcagtggtg
    12001 gaagaacggt ctcagaactg tttgtttcaa ttggccattt aagtttagta gtaaaagact
    12061 ggttaatgat aacaatgcat cgtaaaacct tcagaaggaa aggagaatgt tttgtggacc
    12121 actttggttt tcttttttgc gtgtggcagt tttaagttat tagtttttaa aatcagtact
    12181 ttttaatgga aacaacttga ccaaaaattt gtcacagaat tttgagaccc attaaaaaag
    12241 ttaaatgaga aacctgtgtg ttcctttggt caacaccgag acatttaggt gaaagacatc
    12301 taattctggt tttacgaatc tggaaacttc ttgaaaatgt aattcttgag ttaacacttc
    12361 tgggtggaga atagggttgt tttcccccca cataattgga aggggaagga atatcattta
    12421 aagctatggg agggttgctt tgattacaac actggagaga aatgcagcat gttgctgatt
    12481 gcctgtcact aaaacaggcc aaaaactgag tccttgggtt gcatagaaag ctgccaaacg
    12541 ctagccaaca agctcgtcat cgctttgcag aagagcagag aggatatgct catcgtctaa
    12601 agaactaccc attttattat atattagtca cgatatctat aacaagaaaa tatatatata
    12661 ataagttatc acgtaagtag aacatgaaat aacaatataa ttatcgtatg agttaaatct
    12721 taaaagtcac gtaaaagata atcatgcgtc attttgactc acgcggtcgt tatagttcaa
    12781 aatcagtgac acttaccgca ttgacaagca cgcctcacgg gagctccaag cggcgactga
    12841 gatgtcctaa atgcacagcg acggattcgc gctatttaga aagagagagc aatatttcaa
    12901 gaatgcatgc gtcaatttta cgcagactat ctttctaggg ttaaaaaaga tttgcgcttt
    12961 actcgaccta aactttaaac acgtcataga atcttcgttt gacaaaaacc acattgtggc
    13021 caagctgtgt gacgcgacgc gcgctaaaga atggcaaacc aagtcgcgcg aggtaccagc
    13081 ttttgttccc tttagtgagg gttaatttcg agcttggcgt aatcatggtc atagctgttt
    13141 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
    13201 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
    13261 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
    13321 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
    13381 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
    13441 acagaatcag gggataacgc aggaaagaac atgtgagcaa aagg
//
