LOCUS       Exported               12335 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  All-in-One piggyBac transposon Destination vector for dox-inducible 
            expression of Gateway cloned elements (inducible mCherry and 
            constitutive rtTA and puromycin resistance).
ACCESSION   .
VERSION     .
KEYWORDS    PB-TAG-ERP2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12335)
  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 12335)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..12335
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   711..773
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    1145..1163
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1182..1200
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1219..1237
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1256..1274
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1293..1311
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1330..1348
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1367..1385
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1404..1422
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1444..1468
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    1548..1672
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1697..1727
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1781..2440
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1847..1866)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2782..3087
                     /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     2966..2985
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3128..3252)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    3294..3879
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             3880..4599
                     /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             3880..4596
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3925..3946)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4186..4205)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4533..4554
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4611..4630)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4676..4731
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4730..4749)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     enhancer        5172..5551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             6989..7735
                     /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    7743..8328
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             8329..8928
                     /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(8329..8348)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8825..8845
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    9014..9586
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     repeat_region   9960..9994
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(10144..10162)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10183..10199)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10183..10199)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10196..10218)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10207..10223
                     /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(10231..10261)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10276..10297
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10414..10431)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10585..11173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10665..10684)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11344..12204)
                     /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     11967..11986
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12205..12309)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg cgccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcgg
      661 tattcacgac agcaggctga ataataaaaa aattagaaac tattatttaa ccctagaaag
      721 ataatcatat tgtgacgtac gttaaagata atcatgcgta aaattgacgc atgtgtttta
      781 tcggtctgta tatcgaggtt tatttattaa tttgaataga tattaagttt tattatattt
      841 acacttacat actaataata aattcaacaa acaatttatt tatgtttatt tatttattaa
      901 aaaaaaacaa aaactcaaaa tttcttctat aaagtaacaa aacttttaaa cattctctct
      961 tttacaaaaa taaacttatt ttgtacttta aaaacagtca tgttgtatta taaaataagt
     1021 aattagctta acctatacat aatagaaaca aattatactt attagtcagt cagaaacaac
     1081 tttggcacat atcaatatta tgctctcgtt aattaactcg agggatccaa gctggccgca
     1141 cgtctcccta tcagtgatag agaagtcgac acgtctcgag ctccctatca gtgatagaga
     1201 aggtacgtct agaacgtctc cctatcagtg atagagaagt cgacacgtct cgagctccct
     1261 atcagtgata gagaaggtac gtctagaacg tctccctatc agtgatagag aagtcgacac
     1321 gtctcgagct ccctatcagt gatagagaag gtacgtctag aacgtctccc tatcagtgat
     1381 agagaagtcg acacgtctcg agctccctat cagtgataga gaaggtaccc cctatataag
     1441 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
     1501 ccatagaaga caccgggacc gatccagcct ccgcggtggc ggccatcaca agtttgtaca
     1561 aaaaagctga acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc
     1621 ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcacta tggcggccgc
     1681 attaggcacc ccaggcttta cactttatgc ttccggctcg tataatgtgt ggattttgag
     1741 ttaggatccg tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg
     1801 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
     1861 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
     1921 cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
     1981 gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag
     2041 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag
     2101 tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta
     2161 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc
     2221 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
     2281 cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
     2341 ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga
     2401 attacaacag tactgcgatg agtggcaggg cggggcgtaa acgcgtggat ccggcttact
     2461 aaaagccaga taacagtatg cgtatttgcg cgctgatttt tgcggtataa gaatatatac
     2521 tgatatgtat acccgaagta tgtcaaaaag aggtatgcta tgaagcagcg tattacagtg
     2581 acagttgaca gcgacagcta tcagttgctc aaggcatata tgatgtcaat atctccggtc
     2641 tggtaagcac aaccatgcag aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg
     2701 aaaatcagga agggatggct gaggtcgccc ggtttattga aatgaacggc tcttttgctg
     2761 acgagaacag gggctggtga aatgcagttt aaggtttaca cctataaaag agagagccgt
     2821 tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg acggatggtg
     2881 atccccctgg ccagtgcacg tctgctgtca gataaagtct cccgtgaact ttacccggtg
     2941 gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggtc
     3001 tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc
     3061 attaacctga tgttctgggg aatataaatg tcaggctccc ttatacacag ccagtctgca
     3121 ggtcgaccat agtgactgga tatgttgtgt tttacagtat tatgtagtct gttttttatg
     3181 caaaatctaa tttaatatat tgatatttat atcattttac gtttctcgtt cagctttctt
     3241 gtacaaagtg gtgatctagc taaaacgcgg cctcgaatcg aattccgccc ccccccctct
     3301 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     3361 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     3421 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     3481 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
     3541 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     3601 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     3661 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     3721 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
     3781 tttacatgtg tttagtcgag gttaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     3841 ttttcctttg aaaaacacga tgataatatg gccacaacca tggtgagcaa gggcgaggag
     3901 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     3961 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     4021 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     4081 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     4141 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     4201 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     4261 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     4321 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     4381 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     4441 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     4501 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     4561 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga tctttttccc
     4621 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
     4681 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     4741 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     4801 acatatgcca tatgctggct gccatgaaca aaggtggcta taaagaggtc atcagtatat
     4861 gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat
     4921 tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca
     4981 tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct
     5041 tctcttatga agatccctcg acctgcagga attcgattac ttaagcctag gatccagatc
     5101 tgtacattcg aatcgatgaa ttcgttaacc atggaagctt ctgcagcggc cgcactagcg
     5161 gtggcggcct cgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     5221 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     5281 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     5341 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     5401 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     5461 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     5521 catctacgta ttagtcatcg ctattaccat gatgatgcgg ttttggcagt acatcaatgg
     5581 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     5641 gagtttgttt tgactagtgg agccgagagt aattcataca aaaggaggga tcgccttcgc
     5701 aaggggagag cccagggacc gtccctaaat tctcacagac ccaaatccct gtagccgccc
     5761 cacgacagcg cgaggagcat gcgcccaggg ctgagcgcgg gtagatcaga gcacacaagc
     5821 tcacagtccc cggcggtggg gggaggggcg cgctgagcgg gggccaggga gctggcgcgg
     5881 ggcaaactgg gaaagtggtg tcgtgtgctg gctccgccct cttcccgagg gtgggggaga
     5941 acggtatata agtgcggtag tcgccttgga cgttcttttt cgcaacgggt ttgccgtcag
     6001 aacgcaggtg agtggcgggt gtggcttccg cgggccccgg agctggagcc ctgctctgag
     6061 cgggccgggc tgatatgcga gtgtcgtccg cagggtttag ctgtgagcat tcccacttcg
     6121 agtggcgggc ggtgcggggg tgagagtgcg aggcctagcg gcaaccccgt agcctcgcct
     6181 cgtgtccggc ttgaggccta gcgtggtgtc cgccgccgcg tgccactccg gccgcactat
     6241 gcgttttttg tccttgctgc cctcgattgc cttccagcag catgggctaa caaagggagg
     6301 gtgtggggct cactcttaag gagcccatga agcttacgtt ggataggaat ggaagggcag
     6361 gaggggcgac tggggcccgc ccgccttcgg agcacatgtc cgacgccacc tggatggggc
     6421 gaggcctgtg gctttccgaa gcaatcgggc gtgagtttag cctacctggg ccatgtggcc
     6481 ctagcactgg gcacggtctg gcctggcggt gccgcgttcc cttgcctccc aacaagggtg
     6541 aggccgtccc gcccggcacc agttgcttgc gcggaaagat ggccgctccc ggggccctgt
     6601 tgcaaggagc tcaaaatgga ggacgcggca gcccggtgga gcgggcgggt gagtcaccca
     6661 cacaaaggaa gagggccttg cccctcgccg gccgctgctt cctgtgaccc cgtggtctat
     6721 cggccgcata gtcacctcgg gcttctcttg agcaccgctc gtcgcggcgg ggggagggga
     6781 tctaatggcg ttggagtttg ttcacatttg gtgggtggag actagtcagg ccagcctggc
     6841 gctggaagtc attcttggaa tttgcccctt tgagtttgga gcgaggctaa ttctcaagcc
     6901 tcttagcggt tcaaaggtat tttctaaacc cgtttccagg tgttgtgaaa gccaccgcta
     6961 attcaaagca atccggagta aattcaccat gtctagactg gacaagagca aagtcataaa
     7021 cggcgctctg gaattactca atggagtcgg tatcgaaggc ctgacgacaa ggaaactcgc
     7081 tcaaaagctg ggagttgagc agcctaccct gtactggcac gtgaagaaca agcgggccct
     7141 gctcgatgcc ctgccaatcg agatgctgga caggcatcat acccacttct gccccctgga
     7201 aggcgagtca tggcaagact ttctgcggaa caacgccaag tcattccgct gtgctctcct
     7261 ctcacatcgc gacggggcta aagtgcatct cggcacccgc ccaacagaga aacagtacga
     7321 aaccctggaa aatcagctcg cgttcctgtg tcagcaaggc ttctccctgg agaacgcact
     7381 gtacgctctg tccgccgtgg gccactttac actgggctgc gtattggagg aacaggagca
     7441 tcaagtagca aaagaggaaa gagagacacc taccaccgat tctatgcccc cacttctgag
     7501 acaagcaatt gagctgttcg accggcaggg agccgaacct gccttccttt tcggcctgga
     7561 actaatcata tgtggcctgg agaaacagct aaagtgcgaa agcggcgggc cggccgacgc
     7621 ccttgacgat tttgacttag acatgctccc agccgatgcc cttgacgact ttgaccttga
     7681 tatgctgcct gctgacgctc ttgacgattt tgaccttgac atgctccccg ggtaaaattc
     7741 cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg
     7801 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc
     7861 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa
     7921 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga
     7981 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc
     8041 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc
     8101 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac
     8161 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg
     8221 tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaacg tctaggcccc ccgaaccacg
     8281 gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat gaccgagtac
     8341 aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg caccctcgcc
     8401 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     8461 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     8521 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     8581 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     8641 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     8701 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     8761 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     8821 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     8881 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac
     8941 gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcgaaatg accgaccaag
     9001 cgaattcgct aggattatcc ctaatacctg ccaccccact cttaatcagt ggtggaagaa
     9061 cggtctcaga actgtttgtt tcaattggcc atttaagttt agtagtaaaa gactggttaa
     9121 tgataacaat gcatcgtaaa accttcagaa ggaaaggaga atgttttgtg gaccactttg
     9181 gttttctttt ttgcgtgtgg cagttttaag ttattagttt ttaaaatcag tactttttaa
     9241 tggaaacaac ttgaccaaaa atttgtcaca gaattttgag acccattaaa aaagttaaat
     9301 gagaaacctg tgtgttcctt tggtcaacac cgagacattt aggtgaaaga catctaattc
     9361 tggttttacg aatctggaaa cttcttgaaa atgtaattct tgagttaaca cttctgggtg
     9421 gagaataggg ttgttttccc cccacataat tggaagggga aggaatatca tttaaagcta
     9481 tgggagggtt gctttgatta caacactgga gagaaatgca gcatgttgct gattgcctgt
     9541 cactaaaaca ggccaaaaac tgagtccttg ggttgcatag aaagctgcca aacgctagcc
     9601 aacaagctcg tcatcgcttt gcagaagagc agagaggata tgctcatcgt ctaaagaact
     9661 acccatttta ttatatatta gtcacgatat ctataacaag aaaatatata tataataagt
     9721 tatcacgtaa gtagaacatg aaataacaat ataattatcg tatgagttaa atcttaaaag
     9781 tcacgtaaaa gataatcatg cgtcattttg actcacgcgg tcgttatagt tcaaaatcag
     9841 tgacacttac cgcattgaca agcacgcctc acgggagctc caagcggcga ctgagatgtc
     9901 ctaaatgcac agcgacggat tcgcgctatt tagaaagaga gagcaatatt tcaagaatgc
     9961 atgcgtcaat tttacgcaga ctatctttct agggttaaaa aagatttgcg ctttactcga
    10021 cctaaacttt aaacacgtca tagaatcttc gtttgacaaa aaccacattg tggccaagct
    10081 gtgtgacgcg acgcgcgcta aagaatggca aaccaagtcg cgcgaggtac ccagcttttg
    10141 ttccctttag tgagggttaa ttccgagctt ggcgtaatca tggtcatagc tgtttcctgt
    10201 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
    10261 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
    10321 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
    10381 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
    10441 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
    10501 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
    10561 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
    10621 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
    10681 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
    10741 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
    10801 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
    10861 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
    10921 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
    10981 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
    11041 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
    11101 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
    11161 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
    11221 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
    11281 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
    11341 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
    11401 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
    11461 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
    11521 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
    11581 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
    11641 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
    11701 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
    11761 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
    11821 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
    11881 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
    11941 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
    12001 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
    12061 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
    12121 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
    12181 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
    12241 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
    12301 ggttccgcgc acatttcccc gaaaagtgcc acctg
//
