LOCUS       sequence-484261-       12350 bp    DNA     circular SYN 21-AUG-2025
DEFINITION  All-in-One piggyBac transposon Destination vector for dox-inducible 
            expression of Gateway cloned elements (inducible GFP and 
            constitutive rtTA and puromycin resistance).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12350)
  AUTHORS   Kim SI, Oceguera-Yanez F, Sakurai C, Nakagawa M, Yamanaka S, Woltjen
            K
  TITLE     Inducible Transgene Expression in Human iPS Cells Using Versatile 
            All-in-One piggyBac Transposons.
  JOURNAL   Methods Mol Biol. 2016;1357:111-31. doi: 10.1007/7651_2015_251.
  PUBMED    26025620
REFERENCE   2  (bases 1 to 12350)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 21, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods Mol
            Biol."; date: "2016"; volume: "1357:111-31. doi"; pages: " 
            10.1007/7651_2015_251"
COMMENT     Sequence Label: PB-TAG-ERP2
FEATURES             Location/Qualifiers
     source          1..12350
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(23..478)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(160..181)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     372..391
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     605..627
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     619..636
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     620..636
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     643..662
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        643..661
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   728..790
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    1162..1180
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1199..1217
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1236..1254
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1273..1291
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1310..1328
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1347..1365
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1384..1402
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1421..1439
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1461..1485
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    1565..1689
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1714..1719,1720..1737,1738..1744)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1798..2457
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1864..1883)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2799..3104
                     /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     2983..3002
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3145..3269)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    join(3311..3885,3886..3888,3889..3897)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             join(3898..3900,3901..3903,3904..4617)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3898..3900,3901..3903,3904..4614)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3943..3964)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4204..4223)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4551..4572
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4629..4648)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4694..4749
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4748..4767)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     enhancer        5192..5571
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5686..6991
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    6028..6959
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     CDS             join(7008..7625,7626..7631,7632..7667,7668..7670,7671..7706,
                     7707..7709,7710..7745,7746..7754)
                     /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    join(7781..8355,8356..8358,8359..8367)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             8368..8967
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(8368..8387)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8864..8884
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    9053..9625
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     repeat_region   9999..10033
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(10182..10200)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10221..10237)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10221..10237)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10234..10256)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10245..10261
                     /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(10269..10275,10276..10293,10294..10299))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10314..10335
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10452..10469)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10623..11211)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10703..10722)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11382..12173,12174..12242))
                     /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     12005..12024
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12243..12347)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta
       61 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
      121 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
      181 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
      241 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
      301 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
      361 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
      421 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
      481 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
      541 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
      601 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag
      661 ggcgaattgg agctcggtat tcacgacagc aggctgaata ataaaaaaat tagaaactat
      721 tatttaaccc tagaaagata atcatattgt gacgtacgtt aaagataatc atgcgtaaaa
      781 ttgacgcatg tgttttatcg gtctgtatat cgaggtttat ttattaattt gaatagatat
      841 taagttttat tatatttaca cttacatact aataataaat tcaacaaaca atttatttat
      901 gtttatttat ttattaaaaa aaaacaaaaa ctcaaaattt cttctataaa gtaacaaaac
      961 ttttaaacat tctctctttt acaaaaataa acttattttg tactttaaaa acagtcatgt
     1021 tgtattataa aataagtaat tagcttaacc tatacataat agaaacaaat tatacttatt
     1081 agtcagtcag aaacaacttt ggcacatatc aatattatgc tctcgttaat taactcgagg
     1141 gatccaagct ggccgcacgt ctccctatca gtgatagaga agtcgacacg tctcgagctc
     1201 cctatcagtg atagagaagg tacgtctaga acgtctccct atcagtgata gagaagtcga
     1261 cacgtctcga gctccctatc agtgatagag aaggtacgtc tagaacgtct ccctatcagt
     1321 gatagagaag tcgacacgtc tcgagctccc tatcagtgat agagaaggta cgtctagaac
     1381 gtctccctat cagtgataga gaagtcgaca cgtctcgagc tccctatcag tgatagagaa
     1441 ggtaccccct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
     1501 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg cggtggcggc
     1561 catcacaagt ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa atatcaatat
     1621 attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca
     1681 gtcactatgg cggccgcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     1741 aatgtgtgga ttttgagtta ggatccgtcg agattttcag gagctaagga agctaaaatg
     1801 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat
     1861 tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt
     1921 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
     1981 attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag
     2041 ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg
     2101 ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca catatattcg
     2161 caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat
     2221 atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc
     2281 aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac
     2341 aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc
     2401 ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaaacg
     2461 cgtggatccg gcttactaaa agccagataa cagtatgcgt atttgcgcgc tgatttttgc
     2521 ggtataagaa tatatactga tatgtatacc cgaagtatgt caaaaagagg tatgctatga
     2581 agcagcgtat tacagtgaca gttgacagcg acagctatca gttgctcaag gcatatatga
     2641 tgtcaatatc tccggtctgg taagcacaac catgcagaat gaagcccgtc gtctgcgtgc
     2701 cgaacgctgg aaagcggaaa atcaggaagg gatggctgag gtcgcccggt ttattgaaat
     2761 gaacggctct tttgctgacg agaacagggg ctggtgaaat gcagtttaag gtttacacct
     2821 ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc
     2881 ccgggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtctccc
     2941 gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata
     3001 tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa
     3061 atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca ggctccctta
     3121 tacacagcca gtctgcaggt cgaccatagt gactggatat gttgtgtttt acagtattat
     3181 gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt
     3241 tctcgttcag ctttcttgta caaagtggtg atctagctaa aacgcggcct cgaatcgaat
     3301 tccgcccccc cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
     3361 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     3421 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     3481 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     3541 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     3601 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     3661 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     3721 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     3781 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     3841 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg
     3901 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     3961 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     4021 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     4081 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     4141 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     4201 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     4261 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     4321 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     4381 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     4441 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     4501 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     4561 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc
     4621 ggccgcgatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     4681 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     4741 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     4801 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta
     4861 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     4921 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     4981 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     5041 ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagga attcgattac
     5101 ttaagcctag gatccagatc tgtacattcg aatcgatgaa ttcgttaacc atggaagctt
     5161 ctgcagcggc cgcactagcg gtggcggcct cgacattgat tattgactag ttattaatag
     5221 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     5281 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     5341 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     5401 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     5461 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     5521 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat gatgatgcgg
     5581 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     5641 caccccattg acgtcaatgg gagtttgttt tgactagtgg agccgagagt aattcataca
     5701 aaaggaggga tcgccttcgc aaggggagag cccagggacc gtccctaaat tctcacagac
     5761 ccaaatccct gtagccgccc cacgacagcg cgaggagcat gcgcccaggg ctgagcgcgg
     5821 gtagatcaga gcacacaagc tcacagtccc cggcggtggg gggaggggcg cgctgagcgg
     5881 gggccaggga gctggcgcgg ggcaaactgg gaaagtggtg tcgtgtgctg gctccgccct
     5941 cttcccgagg gtgggggaga acggtatata agtgcggtag tcgccttgga cgttcttttt
     6001 cgcaacgggt ttgccgtcag aacgcaggtg agtggcgggt gtggcttccg cgggccccgg
     6061 agctggagcc ctgctctgag cgggccgggc tgatatgcga gtgtcgtccg cagggtttag
     6121 ctgtgagcat tcccacttcg agtggcgggc ggtgcggggg tgagagtgcg aggcctagcg
     6181 gcaaccccgt agcctcgcct cgtgtccggc ttgaggccta gcgtggtgtc cgccgccgcg
     6241 tgccactccg gccgcactat gcgttttttg tccttgctgc cctcgattgc cttccagcag
     6301 catgggctaa caaagggagg gtgtggggct cactcttaag gagcccatga agcttacgtt
     6361 ggataggaat ggaagggcag gaggggcgac tggggcccgc ccgccttcgg agcacatgtc
     6421 cgacgccacc tggatggggc gaggcctgtg gctttccgaa gcaatcgggc gtgagtttag
     6481 cctacctggg ccatgtggcc ctagcactgg gcacggtctg gcctggcggt gccgcgttcc
     6541 cttgcctccc aacaagggtg aggccgtccc gcccggcacc agttgcttgc gcggaaagat
     6601 ggccgctccc ggggccctgt tgcaaggagc tcaaaatgga ggacgcggca gcccggtgga
     6661 gcgggcgggt gagtcaccca cacaaaggaa gagggccttg cccctcgccg gccgctgctt
     6721 cctgtgaccc cgtggtctat cggccgcata gtcacctcgg gcttctcttg agcaccgctc
     6781 gtcgcggcgg ggggagggga tctaatggcg ttggagtttg ttcacatttg gtgggtggag
     6841 actagtcagg ccagcctggc gctggaagtc attcttggaa tttgcccctt tgagtttgga
     6901 gcgaggctaa ttctcaagcc tcttagcggt tcaaaggtat tttctaaacc cgtttccagg
     6961 tgttgtgaaa gccaccgcta attcaaagca atccggagta attcaccatg tctagactgg
     7021 acaagagcaa agtcataaac ggcgctctgg aattactcaa tggagtcggt atcgaaggcc
     7081 tgacgacaag gaaactcgct caaaagctgg gagttgagca gcctaccctg tactggcacg
     7141 tgaagaacaa gcgggccctg ctcgatgccc tgccaatcga gatgctggac aggcatcata
     7201 cccacttctg ccccctggaa ggcgagtcat ggcaagactt tctgcggaac aacgccaagt
     7261 cattccgctg tgctctcctc tcacatcgcg acggggctaa agtgcatctc ggcacccgcc
     7321 caacagagaa acagtacgaa accctggaaa atcagctcgc gttcctgtgt cagcaaggct
     7381 tctccctgga gaacgcactg tacgctctgt ccgccgtggg ccactttaca ctgggctgcg
     7441 tattggagga acaggagcat caagtagcaa aagaggaaag agagacacct accaccgatt
     7501 ctatgccccc acttctgaga caagcaattg agctgttcga ccggcaggga gccgaacctg
     7561 ccttcctttt cggcctggaa ctaatcatat gtggcctgga gaaacagcta aagtgcgaaa
     7621 gcggcgggcc ggccgacgcc cttgacgatt ttgacttaga catgctccca gccgatgccc
     7681 ttgacgactt tgaccttgat atgctgcctg ctgacgctct tgacgatttt gaccttgaca
     7741 tgctccccgg gtaactaagt aaggatcatt ccgccccccc cccctctccc tccccccccc
     7801 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     7861 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     7921 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     7981 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc
     8041 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     8101 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     8161 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     8221 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt
     8281 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     8341 aaacacgatg ataatatggc cacaaccatg accgagtaca agcccacggt gcgcctcgcc
     8401 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     8461 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     8521 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     8581 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     8641 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     8701 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     8761 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     8821 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     8881 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     8941 tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa
     9001 aggagcgcac gaccccatgc atcgaaatga ccgaccaagc gaattcgcta ggattatccc
     9061 taatacctgc caccccactc ttaatcagtg gtggaagaac ggtctcagaa ctgtttgttt
     9121 caattggcca tttaagttta gtagtaaaag actggttaat gataacaatg catcgtaaaa
     9181 ccttcagaag gaaaggagaa tgttttgtgg accactttgg ttttcttttt tgcgtgtggc
     9241 agttttaagt tattagtttt taaaatcagt actttttaat ggaaacaact tgaccaaaaa
     9301 tttgtcacag aattttgaga cccattaaaa aagttaaatg agaaacctgt gtgttccttt
     9361 ggtcaacacc gagacattta ggtgaaagac atctaattct ggttttacga atctggaaac
     9421 ttcttgaaaa tgtaattctt gagttaacac ttctgggtgg agaatagggt tgttttcccc
     9481 ccacataatt ggaaggggaa ggaatatcat ttaaagctat gggagggttg ctttgattac
     9541 aacactggag agaaatgcag catgttgctg attgcctgtc actaaaacag gccaaaaact
     9601 gagtccttgg gttgcataga aagctgccaa acgctagcca acaagctcgt catcgctttg
     9661 cagaagagca gagaggatat gctcatcgtc taaagaacta cccattttat tatatattag
     9721 tcacgatatc tataacaaga aaatatatat ataataagtt atcacgtaag tagaacatga
     9781 aataacaata taattatcgt atgagttaaa tcttaaaagt cacgtaaaag ataatcatgc
     9841 gtcattttga ctcacgcggt cgttatagtt caaaatcagt gacacttacc gcattgacaa
     9901 gcacgcctca cgggagctcc aagcggcgac tgagatgtcc taaatgcaca gcgacggatt
     9961 cgcgctattt agaaagagag agcaatattt caagaatgca tgcgtcaatt ttacgcagac
    10021 tatctttcta gggttaaaaa agatttgcgc tttactcgac ctaaacttta aacacgtcat
    10081 agaatcttcg tttgacaaaa accacattgt ggccaagctg tgtgacgcga cgcgcgctaa
    10141 agaatggcaa accaagtcgc gcgaggtacc agcttttgtt ccctttagtg agggttaatt
    10201 tcgagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
    10261 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
    10321 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
    10381 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
    10441 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
    10501 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
    10561 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
    10621 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
    10681 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
    10741 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
    10801 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
    10861 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
    10921 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
    10981 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
    11041 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
    11101 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
    11161 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
    11221 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
    11281 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
    11341 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
    11401 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
    11461 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
    11521 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
    11581 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
    11641 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
    11701 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
    11761 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
    11821 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
    11881 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
    11941 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
    12001 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
    12061 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
    12121 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
    12181 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
    12241 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
    12301 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
//
