LOCUS       Exported                8664 bp ds-DNA     circular SYN 08-JUN-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8664)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 5, 2023 from SnapGene 4.0.8
            http://www.snapgene.com
COMMENT     Notes o The plasmid has been transformed in SURE2 cells 
            (Stratagene), to avoid recombination. o The cassette 
            [5'TR-MCS-pA-3'TR] has been cloned between the SacII and KpnI sites 
            of the vector pBluescript II SK+. It can be removed by cutting 
            SacII-KpnI or by cutting with AscI. o The 5'TR contains the two 
            point mutations described in Lacoste et al., 2009 (Cell Stem Cell). 
            o The following primers can be used to sequence the plasmid: M13-rev
            and T3 (upstream of the 5'TR, top strand); M13-fw and T7 (downstream
            of the 3'TR, bottom strand). o Any restriction site can be used to 
            clone your favorite promoter/gene. However, I would suggest cloning 
            the promoter between XhoI-BamHI and the gene between BamHI-NotI, for
            consistency with previous constructs by Arnaud and others in the 
            lab. o You don't need to add a polyAdenylation (pA) sequence, since 
            it is already present in the vector. If undesired, the pA can be 
            removed with the EcoRI site.
FEATURES             Location/Qualifiers
     source          1..8664
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          326..557
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     source          2695..2757
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     source          4858..5088
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     repeat_region   1..325
                     /label=5' PB Terminal Repeat (ePB)
                     /note="5' Piggybac Transposon Terminal Repeat. Optimized 
                     for human IPSC by Arnaud et al. 2009."
     misc_feature    14..26
                     /label=13 bp terminal repeat (TR)
     misc_feature    27..29
                     /label=3 bp spacer
     misc_feature    30..48
                     /label=19 bp Internal Repeat (IR)
     enhancer        326..557
                     /label=core Insulator
                     /label=core insul enhancer
                     /label=core insul
     misc_feature    564..590
                     /label=I-CeuI site
                     /label=ICeuI site
                     /note="ICeuI site"
     misc_feature    591..605
                     /label=AarI site
     promoter        612..861
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA. 
                     Taken from Feng Zhang lab CRISPR sgRNA plasmids (Addgene 
                     42230, 60958, 61591)."
     primer_bind     612..632
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     783..802
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     regulatory      832..837
                     /regulatory_class="TATA_box"
     misc_signal     861
                     /label=TSS +1
                     /note="Transcription start site for human U6 promoter. 
                     First base of transcript. G is preferred."
     regulatory      881..886
                     /regulatory_class="terminator"
     misc_feature    887..901
                     /label=AarI site
     misc_feature    894..911
                     /label=ISceI site
                     /note="ISceI site"
     misc_feature    912..1455
                     /label=EF1 promoter
                     /note="/ApEinfo_fwdcolor=#ff80c0
                     /ApEinfo_revcolor=green"
     promoter        942..1153
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1166..1434
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      1468..1477
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1474..2694
                     /codon_start=1
                     /label=EPN-11
                     /translation="MGCIKSKRKDNLNLQSRPEPTAPPEESFRSGVETTTPPQKQEPID
                     KELYPLTSLRSLFGNDPSSQKIEELFKKHKIVAVLRANSVEEAKKKALAVFLGGVHLIE
                     ITFTVPDADTVIKELSFLKEMGAIIGAGTVTSVEQCRKAVESGAEFIVSPHLDEEISQF
                     CKEKGVFYMPGVMTPTELVKAMKLGHTILKLFPGEVVGPQFVKAMKGPFPNVKFVPTGG
                     VNLDNVCEWFKAGVLAVGVGSALVKGTPVEVAEKAKAFVEKIRGCTASNFTQFVLVDNG
                     GTGDVTVAPSNFANGVAEWISSNSRSQAYKVTCSVRQSSAQKRKYTIKVEVPKVATQTV
                     GGVELPVAAWRSYLNMELTIPIFATNSDCELIVKAMQGLLKDGNPIPSAIAANSGIYEQ
                     KLISEEDL"
     CDS             1477..1512
                     /codon_start=1
                     /product="Myristoylation/palmitoylation motif from Lyn 
                     kinase (residues 2-13)"
                     /label=Lyn
                     /translation="GCIKSKRKDNLN"
     CDS             1513..1668
                     /codon_start=1
                     /product="Plays a role in budding of the assembled particle
                     by interacting with the host class E VPS proteins TSG101 
                     and PDCD6IP/AIP1."
                     /label=HIV-1 p6
                     /translation="LQSRPEPTAPPEESFRSGVETTTPPQKQEPIDKELYPLTSLRSLF
                     GNDPSSQ"
     CDS             1531..1542
                     /product="Late-budding domains (L domains) are short 
                     sequence motifs essential for viral particle budding. They 
                     recruit proteins of the host ESCRT machinery (Endosomal 
                     Sorting Complex Required for Transport) or ESCRT-associated
                     proteins. p6-gag contains two L domains: a PTAP/PSAP motif,
                     which interacts with the UEV domain of TSG101 and a 
                     LYPX(n)L motif which interacts with PDCD6IP/AIP1 
                     (https://www.uniprot.org/uniprot/P04591). Labeled 
                     ""ESCRT-I"" in Votteler et al. 2016, Extended Data Figure 
                     1a."
                     /label=HIV-1 p6-gag L domain
     CDS             1615..1644
                     /product="Late-budding domains (L domains) are short 
                     sequence motifs essential for viral particle budding. They 
                     recruit proteins of the host ESCRT machinery (Endosomal 
                     Sorting Complex Required for Transport) or ESCRT-associated
                     proteins. p6-gag contains two L domains: a PTAP/PSAP motif,
                     which interacts with the UEV domain of TSG101 and a 
                     LYPX(n)L motif which interacts with PDCD6IP/AIP1 
                     (https://www.uniprot.org/uniprot/P04591). Labeled ""ALIX"" 
                     in Votteler et al. 2016, Extended Data Figure 1a."
                     /label=HIV-1 p6-gag L domain
     CDS             1669..2277
                     /codon_start=1
                     /product="I3-01 Interaction domain"
                     /label=I3-01
                     /translation="KIEELFKKHKIVAVLRANSVEEAKKKALAVFLGGVHLIEITFTVP
                     DADTVIKELSFLKEMGAIIGAGTVTSVEQCRKAVESGAEFIVSPHLDEEISQFCKEKGV
                     FYMPGVMTPTELVKAMKLGHTILKLFPGEVVGPQFVKAMKGPFPNVKFVPTGGVNLDNV
                     CEWFKAGVLAVGVGSALVKGTPVEVAEKAKAFVEKIRGCT"
     CDS             1918..1920
                     /label=Recoded to remove EcoRI
     CDS             2278..2664
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2-N55K
                     /note="binds with high affinity to a specific stem-loop 
                     structure in the viral RNA (Lim et al., 1994)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     CDS             2665..2694
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2695..2757
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="GSGEGRGSLLTCGDVEENPGP"
     CDS             2758..3474
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     polyA_signal    3489..3725
                     /label=pA
                     /note="pA"
     polyA_signal    3602..3723
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    3734..4183
                     /label=PGK promoter
                     /label=PGK(-)
                     /note="/ApEinfo_fwdcolor=#ff8000
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
                     /ApEinfo_revcolor=#ff8040"
     regulatory      4193..4202
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4199..4597
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BlastR
                     /note="BSD
                     confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4615..4839
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        complement(4858..5088)
                     /label=core Insulator
                     /label=core insul(1) enhancer
                     /label=core insul(1)
     repeat_region   5089..5341
                     /label=3' PB Terminal Repeat (ePB)
                     /note="3' Piggybac Transposon Terminal Repeat. Optimized 
                     for human IPSC by Arnaud et al. 2009."
     misc_feature    5266..5284
                     /label=19 bp Internal Repeat (IR)
     misc_feature    5285..5315
                     /label=31 bp spacer
     misc_feature    5316..5328
                     /label=13 bp terminal repeat (TR)
     primer_bind     complement(5827..5843)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5851..5867
                     /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(5875..5905)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5920..5941
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6229..6817)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6988..7782)
                     /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"
     promoter        complement(7783..7887)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7913..8368)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     8510..8526
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gcagctagat taaccctaga aagatagtct gcgtaaaatt gacgcatgca ttcttgaaat
       61 attgctctct ctttctaaat agcgcgaatc cgtcgctgtg catttaggac atctcagtcg
      121 ccgcttggag ctcccgtgag gcgtgcttgt caatgcggta agtgtcactg attttgaact
      181 ataacaaccg cgtgagtcaa aatgacgcat gattatcttt tacgtgactt ttaagattta
      241 actcatacga taattatatt gttatttcgt gttctactta cgtgataact tattatatat
      301 atattttctt gttatagata tccttgaggg acagcccccc cccaaagccc ccagggatgt
      361 aattacgtcc ctcccccgct agggggcagc agcgagccgc ccggggctcc gctccggtcc
      421 ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga cagcccgggc acggggaagg
      481 tggcacggga tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg cagacacctg
      541 gggggatacg gggaaaaatc gattaactat aacggtccta aggtagcgaa cacctgcgct
      601 cgagcgacgt cgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
      661 ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata
      721 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
      781 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
      841 tgtggaaagg acgaaacacc gaccaggtgg gcccacgcgt ttttttcacc tgcattaccc
      901 tgttatccct aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc
      961 cacagtcccc gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg
     1021 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     1081 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     1141 cgccagaaca cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc
     1201 tacctgaggc cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc
     1261 ctcctgaact gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg
     1321 tccggcgctc ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct
     1381 gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct
     1441 gtgaccggcg cctacgctag cggatccgcc accatgggtt gtatcaagag caagcgcaag
     1501 gataatttga acctgcagtc tcgtccggaa ccgaccgcac cgccggaaga atctttccgt
     1561 tctggtgttg aaaccaccac gccaccgcaa aaacaggagc cgatcgacaa agaactgtac
     1621 ccgctgacct ctctgcgttc tctgttcggt aacgacccgt cttctcagaa gatcgaagag
     1681 ctgttcaaga aacacaagat cgttgccgtg ctgcgtgcca atagtgtgga agaagcgaaa
     1741 aagaaagcgc tggcggtttt cctgggcggc gttcatctga ttgaaattac ctttaccgtg
     1801 ccggatgcgg ataccgtgat taaggaactg agctttctga aggaaatggg cgcgattatt
     1861 ggtgcgggca ccgtgaccag cgtggagcag tgccgtaaag cggtggaaag tggcgccgag
     1921 ttcattgtga gtccgcacct ggacgaggaa attagccaat tttgcaagga gaagggtgtg
     1981 ttctatatgc caggcgttat gaccccgacc gaactggtga aagccatgaa actgggccat
     2041 accatcttaa aactgtttcc gggtgaggtg gtgggtccgc agtttgttaa agcgatgaaa
     2101 ggtccgtttc cgaatgtgaa atttgtgcca accggcggtg ttaatctgga caatgtgtgc
     2161 gaatggttca aagcgggcgt gctggccgtg ggcgtgggca gcgcgttagt gaaaggcacc
     2221 ccggtggaag tggcggaaaa ggccaaggcg ttcgttgaga agattcgtgg ctgcaccgct
     2281 tcaaacttta ctcagttcgt gctcgtggac aatggtggga caggggatgt gacagtggct
     2341 ccttctaatt tcgctaatgg ggtggcagag tggatcagct ccaactcacg gagccaggcc
     2401 tacaaggtga catgcagcgt caggcagtct agtgcccaga agagaaagta taccatcaag
     2461 gtggaggtcc ccaaagtggc tacccagaca gtgggcggag tcgaactgcc tgtcgccgct
     2521 tggaggtcct acctgaacat ggagctcact atcccaattt tcgctaccaa ttctgactgt
     2581 gaactcatcg tgaaggcaat gcaggggctc ctcaaagacg gtaatcctat cccttccgcc
     2641 atcgccgcta actcaggtat ctacgaacag aagctgatct ccgaagaaga cctgggatcg
     2701 ggagagggca gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc tggcccaatg
     2761 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     2821 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     2881 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2941 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     3001 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     3061 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     3121 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     3181 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     3241 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     3301 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     3361 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     3421 gagttcgtga ccgccgccgg gatcactcac ggcatggacg agctgtacaa gtaagcggcc
     3481 gcgtcgacga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     3541 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     3601 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3661 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3721 ttaagcgccg gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta
     3781 ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag
     3841 tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag
     3901 cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc
     3961 tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa
     4021 ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt
     4081 cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct
     4141 cttcctcatc tccgggcctt tcgacctgca gcccaagctt accatgggta ccgccaccat
     4201 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
     4261 cagcatcccc atctctgaag attacagcgt cgccagcgca gctctctcta gcgacggccg
     4321 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
     4381 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
     4441 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
     4501 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
     4561 tgaattgctg ccctctggtt atgtgtggga gggctaaggt cacccgcgcg cggactgtgc
     4621 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     4681 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     4741 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     4801 acaatagcag gcatgctggg gatgcggtgg gctctatggt tcgaaccagt acgatggttt
     4861 tccccgtatc cccccaggtg tctgcaggct caaagagcag cgagaagcgt tcagaggaaa
     4921 gcgatcccgt gccaccttcc ccgtgcccgg gctgtccccg cacgctgccg gctcggggat
     4981 gcggggggag cgccggaccg gagcggagcc ccgggcggct cgctgctgcc ccctagcggg
     5041 ggagggacgt aattacatcc ctgggggctt tggggggggg ctgtccctga taaaagtttt
     5101 gttactttat agaagaaatt ttgagttttt gttttttttt aataaataaa taaacataaa
     5161 taaattgttt gttgaattta ttattagtat gtaagtgtaa atataataaa acttaatatc
     5221 tattcaaatt aataaataaa cctcgatata cagaccgata aaacacatgc gtcaatttta
     5281 cgcatgatta tctttaacgt acgtcacaat atgattatct ttctagggtt aatctagctg
     5341 cggcgcgcca tcaggatcat atcgtcgggt cttttttccg gctcagtcat cgcccaagct
     5401 ggcgctatct gggcatcggg gaggaagaag cccgtgcctt ttcccgcgag gttgaagcgg
     5461 catggaaaga gtttgccgag gatgactgct gctgcattga cgttgagcga aaacgcacgt
     5521 ttaccatgat gattcgggaa ggtgtggcca tgcacgcctt taacggtgaa ctgttcgttc
     5581 aggccacctg ggataccagt tcgtcgcggc ttttccggac acagttccgg atggtcagcc
     5641 cgaagcgcat cagcaacccg aacaataccg gcgacagccg gaactgccgt gccggtgtgc
     5701 agattaatga cagcggtgcg gcgctgggat attacgtcag cgaggacggg tatcctggct
     5761 ggatgccgca gaaatggaca tggatacccc gtgagttacc cggcgggcgc gcttggcgta
     5821 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5881 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5941 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6001 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6061 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6121 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6181 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6241 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6301 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6361 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6421 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6481 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6541 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6601 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6661 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6721 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6781 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6841 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6901 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6961 tatatatgag taaacttggt ctgacagtca gaagaactcg tcaagaaggc gatagaaggc
     7021 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc
     7081 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc
     7141 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt
     7201 cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt
     7261 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg
     7321 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg
     7381 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat
     7441 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc
     7501 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac
     7561 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc
     7621 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc
     7681 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca
     7741 agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atactcttcc tttttcaata
     7801 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7861 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta
     7921 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     7981 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     8041 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     8101 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     8161 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     8221 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     8281 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     8341 gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg
     8401 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     8461 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     8521 gccagtgagc gcgcctcgtt cattcacgtt tttgaacccg tggaggacgg gcagactcgc
     8581 ggtgcaaatg tgttttacag cgtgatggag cagatgaaga tgctcgacac gctgcagaac
     8641 acgagctcca ccgcggggcg cgcc
//
