LOCUS       Exported                8631 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 8631)
  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..8631
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          326..557
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     source          2662..2724
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     source          4825..5055
                     /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..2661
                     /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..2661
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2662..2724
                     /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             2725..3441
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     polyA_signal    3456..3692
                     /label=pA
                     /note="pA"
     polyA_signal    3569..3690
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    3701..4150
                     /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      4160..4169
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4166..4564
                     /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    4582..4806
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        complement(4825..5055)
                     /label=core Insulator
                     /label=core insul(1) enhancer
                     /label=core insul(1)
     repeat_region   5056..5308
                     /label=3' PB Terminal Repeat (ePB)
                     /note="3' Piggybac Transposon Terminal Repeat. Optimized 
                     for human IPSC by Arnaud et al. 2009."
     misc_feature    5233..5251
                     /label=19 bp Internal Repeat (IR)
     misc_feature    5252..5282
                     /label=31 bp spacer
     misc_feature    5283..5295
                     /label=13 bp terminal repeat (TR)
     primer_bind     complement(5794..5810)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5818..5834
                     /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(5842..5872)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5887..5908
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6196..6784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6955..7749)
                     /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(7750..7854)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7880..8335)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     8477..8493
                     /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 cgaacagaag
     2641 ctgatctccg aagaagacct gggatcggga gagggcagag gaagtctgct aacatgcggt
     2701 gacgtcgagg agaatcctgg cccaatgagc aagggcgagg agctgttcac cggggtggtg
     2761 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2821 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2881 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2941 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     3001 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     3061 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     3121 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     3181 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     3241 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     3301 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     3361 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactcacggc
     3421 atggacgagc tgtacaagta agcggccgcg tcgacgactc tagatcataa tcagccatac
     3481 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3541 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     3601 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3661 tggtttgtcc aaactcatca atgtatctta agcgccggcg ctacacaagt ggcctctggc
     3721 ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct
     3781 tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt
     3841 cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca
     3901 ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct
     3961 tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc
     4021 tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa
     4081 agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcagcc
     4141 caagcttacc atgggtaccg ccaccatggc caagcctttg tctcaagaag aatccaccct
     4201 cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagatt acagcgtcgc
     4261 cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac
     4321 tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa
     4381 cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg
     4441 gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga
     4501 tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg
     4561 ctaaggtcac ccgcgcgcgg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     4621 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     4681 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     4741 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     4801 ctatggttcg aaccagtacg atggttttcc ccgtatcccc ccaggtgtct gcaggctcaa
     4861 agagcagcga gaagcgttca gaggaaagcg atcccgtgcc accttccccg tgcccgggct
     4921 gtccccgcac gctgccggct cggggatgcg gggggagcgc cggaccggag cggagccccg
     4981 ggcggctcgc tgctgccccc tagcggggga gggacgtaat tacatccctg ggggctttgg
     5041 gggggggctg tccctgataa aagttttgtt actttataga agaaattttg agtttttgtt
     5101 tttttttaat aaataaataa acataaataa attgtttgtt gaatttatta ttagtatgta
     5161 agtgtaaata taataaaact taatatctat tcaaattaat aaataaacct cgatatacag
     5221 accgataaaa cacatgcgtc aattttacgc atgattatct ttaacgtacg tcacaatatg
     5281 attatctttc tagggttaat ctagctgcgg cgcgccatca ggatcatatc gtcgggtctt
     5341 ttttccggct cagtcatcgc ccaagctggc gctatctggg catcggggag gaagaagccc
     5401 gtgccttttc ccgcgaggtt gaagcggcat ggaaagagtt tgccgaggat gactgctgct
     5461 gcattgacgt tgagcgaaaa cgcacgttta ccatgatgat tcgggaaggt gtggccatgc
     5521 acgcctttaa cggtgaactg ttcgttcagg ccacctggga taccagttcg tcgcggcttt
     5581 tccggacaca gttccggatg gtcagcccga agcgcatcag caacccgaac aataccggcg
     5641 acagccggaa ctgccgtgcc ggtgtgcaga ttaatgacag cggtgcggcg ctgggatatt
     5701 acgtcagcga ggacgggtat cctggctgga tgccgcagaa atggacatgg ataccccgtg
     5761 agttacccgg cgggcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5821 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5881 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5941 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6001 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6061 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6121 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6181 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6241 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6301 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6361 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6421 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6481 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6541 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6601 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     6661 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6721 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6781 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6841 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6901 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagtcagaa
     6961 gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta
     7021 aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc
     7081 caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga
     7141 aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag
     7201 atcctcgccg tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc
     7261 ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc
     7321 tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg
     7381 cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga
     7441 caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac
     7501 aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc
     7561 ctcgtcttgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg
     7621 cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca
     7681 gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg
     7741 ttcaatcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7801 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7861 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     7921 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     7981 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     8041 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     8101 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     8161 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     8221 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     8281 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     8341 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     8401 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     8461 tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cctcgttcat tcacgttttt
     8521 gaacccgtgg aggacgggca gactcgcggt gcaaatgtgt tttacagcgt gatggagcag
     8581 atgaagatgc tcgacacgct gcagaacacg agctccaccg cggggcgcgc c
//
