LOCUS       Exported                9018 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 9018)
  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..9018
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          326..557
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     source          3049..3111
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     source          5212..5442
                     /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..1476
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1474..3048
                     /label=EPN-24
     CDS             1474..1866
                     /codon_start=1
                     /product="PLC delta 11-140 (PH) membrane interaction 
                     domain."
                     /label=PLCD (PH)
                     /translation="MHGLQDDPDLQALLKGSQLLKVKSSSWRRERFYKLQEDCKTIWQE
                     SRKVMRSPESQLFSIEDIQEVRMGHRTEGLEKFARDIPEDRCFSIVFKDQRNTLDLIAP
                     SPADAQHWVQGLRKIIHHSGSMDQRQK"
     CDS             1867..2022
                     /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             1885..1896
                     /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             1969..1998
                     /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             2023..2631
                     /codon_start=1
                     /product="I3-01 Interaction domain"
                     /label=I3-01
                     /translation="KIEELFKKHKIVAVLRANSVEEAKKKALAVFLGGVHLIEITFTVP
                     DADTVIKELSFLKEMGAIIGAGTVTSVEQCRKAVESGAEFIVSPHLDEEISQFCKEKGV
                     FYMPGVMTPTELVKAMKLGHTILKLFPGEVVGPQFVKAMKGPFPNVKFVPTGGVNLDNV
                     CEWFKAGVLAVGVGSALVKGTPVEVAEKAKAFVEKIRGCT"
     misc_feature    2274
                     /label=Recoded to remove EcoRI
     CDS             2632..3018
                     /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             3019..3048
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3049..3111
                     /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             3112..3828
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     polyA_signal    3843..4079
                     /label=pA
                     /note="pA"
     polyA_signal    3956..4077
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    4088..4537
                     /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      4547..4556
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4553..4951
                     /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    4969..5193
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        complement(5212..5442)
                     /label=core Insulator
                     /label=core insul(1) enhancer
                     /label=core insul(1)
     repeat_region   5443..5695
                     /label=3' PB Terminal Repeat (ePB)
                     /note="3' Piggybac Transposon Terminal Repeat. Optimized 
                     for human IPSC by Arnaud et al. 2009."
     misc_feature    5620..5638
                     /label=19 bp Internal Repeat (IR)
     misc_feature    5639..5669
                     /label=31 bp spacer
     misc_feature    5670..5682
                     /label=13 bp terminal repeat (TR)
     primer_bind     complement(6181..6197)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6205..6221
                     /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(6229..6259)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6274..6295
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6583..7171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7342..8136)
                     /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(8137..8241)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8267..8722)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     8864..8880
                     /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 accatgcacg gtctgcagga cgacccggac
     1501 ctgcaagcac tgctcaaggg cagccagctg ctgaaagtta aatcttcttc ttggcgtcgt
     1561 gaacgtttct acaaactcca ggaggactgc aaaaccatct ggcaggaatc ccgtaaagtt
     1621 atgcgttctc cggaatctca gctgttctct atcgaagaca tccaggaagt tcgtatgggt
     1681 caccgtaccg aaggtctgga aaaattcgcg cgtgacatcc cggaagaccg ttgcttctcc
     1741 atcgttttca aagaccagcg taacaccctg gacctgatcg cgccgtctcc ggcggacgcg
     1801 cagcactggg ttcagggtct gcgtaaaatc atccaccact ctggttctat ggaccaacgt
     1861 cagaaactgc agtctcgtcc ggaaccgacc gcaccgccgg aagaatcttt ccgttctggt
     1921 gttgaaacca ccacgccacc gcaaaaacag gagccgatcg acaaagaact gtacccgctg
     1981 acctctctgc gttctctgtt cggtaacgac ccgtcttctc agaagatcga agagctgttc
     2041 aagaaacaca agatcgttgc cgtgctgcgt gccaatagtg tggaagaagc gaaaaagaaa
     2101 gcgctggcgg ttttcctggg cggcgttcat ctgattgaaa ttacctttac cgtgccggat
     2161 gcggataccg tgattaagga actgagcttt ctgaaggaaa tgggcgcgat tattggtgcg
     2221 ggcaccgtga ccagcgtgga gcagtgccgt aaagcggtgg aaagtggcgc cgagttcatt
     2281 gtgagtccgc acctggacga ggaaattagc caattttgca aggagaaggg tgtgttctat
     2341 atgccaggcg ttatgacccc gaccgaactg gtgaaagcca tgaaactggg ccataccatc
     2401 ttaaaactgt ttccgggtga ggtggtgggt ccgcagtttg ttaaagcgat gaaaggtccg
     2461 tttccgaatg tgaaatttgt gccaaccggc ggtgttaatc tggacaatgt gtgcgaatgg
     2521 ttcaaagcgg gcgtgctggc cgtgggcgtg ggcagcgcgt tagtgaaagg caccccggtg
     2581 gaagtggcgg aaaaggccaa ggcgttcgtt gagaagattc gtggctgcac cgcttcaaac
     2641 tttactcagt tcgtgctcgt ggacaatggt gggacagggg atgtgacagt ggctccttct
     2701 aatttcgcta atggggtggc agagtggatc agctccaact cacggagcca ggcctacaag
     2761 gtgacatgca gcgtcaggca gtctagtgcc cagaagagaa agtataccat caaggtggag
     2821 gtccccaaag tggctaccca gacagtgggc ggagtcgaac tgcctgtcgc cgcttggagg
     2881 tcctacctga acatggagct cactatccca attttcgcta ccaattctga ctgtgaactc
     2941 atcgtgaagg caatgcaggg gctcctcaaa gacggtaatc ctatcccttc cgccatcgcc
     3001 gctaactcag gtatctacga acagaagctg atctccgaag aagacctggg atcgggagag
     3061 ggcagaggaa gtctgctaac atgcggtgac gtcgaggaga atcctggccc aatgagcaag
     3121 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     3181 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     3241 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     3301 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     3361 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     3421 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     3481 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     3541 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     3601 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     3661 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     3721 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     3781 gtgaccgccg ccgggatcac tcacggcatg gacgagctgt acaagtaagc ggccgcgtcg
     3841 acgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3901 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3961 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4021 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagc
     4081 gccggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc
     4141 aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga
     4201 agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc
     4261 tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg
     4321 gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg
     4381 ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg
     4441 gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct
     4501 catctccggg cctttcgacc tgcagcccaa gcttaccatg ggtaccgcca ccatggccaa
     4561 gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat
     4621 ccccatctct gaagattaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt
     4681 cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg
     4741 cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa
     4801 caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg
     4861 gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt
     4921 gctgccctct ggttatgtgt gggagggcta aggtcacccg cgcgcggact gtgccttcta
     4981 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     5041 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     5101 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     5161 gcaggcatgc tggggatgcg gtgggctcta tggttcgaac cagtacgatg gttttccccg
     5221 tatcccccca ggtgtctgca ggctcaaaga gcagcgagaa gcgttcagag gaaagcgatc
     5281 ccgtgccacc ttccccgtgc ccgggctgtc cccgcacgct gccggctcgg ggatgcgggg
     5341 ggagcgccgg accggagcgg agccccgggc ggctcgctgc tgccccctag cgggggaggg
     5401 acgtaattac atccctgggg gctttggggg ggggctgtcc ctgataaaag ttttgttact
     5461 ttatagaaga aattttgagt ttttgttttt ttttaataaa taaataaaca taaataaatt
     5521 gtttgttgaa tttattatta gtatgtaagt gtaaatataa taaaacttaa tatctattca
     5581 aattaataaa taaacctcga tatacagacc gataaaacac atgcgtcaat tttacgcatg
     5641 attatcttta acgtacgtca caatatgatt atctttctag ggttaatcta gctgcggcgc
     5701 gccatcagga tcatatcgtc gggtcttttt tccggctcag tcatcgccca agctggcgct
     5761 atctgggcat cggggaggaa gaagcccgtg ccttttcccg cgaggttgaa gcggcatgga
     5821 aagagtttgc cgaggatgac tgctgctgca ttgacgttga gcgaaaacgc acgtttacca
     5881 tgatgattcg ggaaggtgtg gccatgcacg cctttaacgg tgaactgttc gttcaggcca
     5941 cctgggatac cagttcgtcg cggcttttcc ggacacagtt ccggatggtc agcccgaagc
     6001 gcatcagcaa cccgaacaat accggcgaca gccggaactg ccgtgccggt gtgcagatta
     6061 atgacagcgg tgcggcgctg ggatattacg tcagcgagga cgggtatcct ggctggatgc
     6121 cgcagaaatg gacatggata ccccgtgagt tacccggcgg gcgcgcttgg cgtaatcatg
     6181 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6241 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6301 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6361 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6421 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6481 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6541 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6601 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6661 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6721 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6781 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6841 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6901 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6961 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7021 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7081 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7141 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7201 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7261 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7321 tgagtaaact tggtctgaca gtcagaagaa ctcgtcaaga aggcgataga aggcgatgcg
     7381 ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc
     7441 aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc
     7501 cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa
     7561 gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct
     7621 ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac
     7681 aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa
     7741 tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac
     7801 tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag
     7861 cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt
     7921 cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg caccggacag
     7981 gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc
     8041 agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc
     8101 cggagaacct gcgtgcaatc catcttgttc aatcatactc ttcctttttc aatattattg
     8161 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8221 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     8281 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     8341 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     8401 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     8461 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     8521 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     8581 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     8641 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     8701 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
     8761 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     8821 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     8881 gagcgcgcct cgttcattca cgtttttgaa cccgtggagg acgggcagac tcgcggtgca
     8941 aatgtgtttt acagcgtgat ggagcagatg aagatgctcg acacgctgca gaacacgagc
     9001 tccaccgcgg ggcgcgcc
//
